Structured Review

Developmental Studies Hybridoma Bank mouse anti fasciclin ii
Neuron-specific depletion of Edis causes abnormal axonogenesis and neurodevelopment. (A) Confocal images of dorsal anterior regions of adult brains with neuron-specific expression of control shgfp (left panel) or shEdis (middle and right panels) driven by Elav-Gal4 driver. In control brain (left panel), <t>anti-FasII</t> antibody delineates the central complex (CC) as well as the vertical α and horizontal β and γ lobes of the mushroom bodies ( MB s), with γ lobes showing weaker FasII signal, as indicated. Depletion of Edis resulted in a spectrum of severe, age-dependent morphological defects in the MBs. In middle panel, one α lobe and one β lobe were mostly missing, and the remaining α and β lobes showed abnormal morphology. In right panel, both α lobes were largely missing, while the two β lobes crossed the midline and merged together. Scale bar: 20 μm. (B) Quantification of MB morphology phenotypes shown in A (Chi-squared test, sample numbers are shown on top, *** p
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Images

1) Product Images from "The circular RNA Edis regulates neurodevelopment and innate immunityA circular RNA Edis-Relish-castor axis regulates neuronal development in Drosophila"

Article Title: The circular RNA Edis regulates neurodevelopment and innate immunityA circular RNA Edis-Relish-castor axis regulates neuronal development in Drosophila

Journal: PLOS Genetics

doi: 10.1371/journal.pgen.1010429

Neuron-specific depletion of Edis causes abnormal axonogenesis and neurodevelopment. (A) Confocal images of dorsal anterior regions of adult brains with neuron-specific expression of control shgfp (left panel) or shEdis (middle and right panels) driven by Elav-Gal4 driver. In control brain (left panel), anti-FasII antibody delineates the central complex (CC) as well as the vertical α and horizontal β and γ lobes of the mushroom bodies ( MB s), with γ lobes showing weaker FasII signal, as indicated. Depletion of Edis resulted in a spectrum of severe, age-dependent morphological defects in the MBs. In middle panel, one α lobe and one β lobe were mostly missing, and the remaining α and β lobes showed abnormal morphology. In right panel, both α lobes were largely missing, while the two β lobes crossed the midline and merged together. Scale bar: 20 μm. (B) Quantification of MB morphology phenotypes shown in A (Chi-squared test, sample numbers are shown on top, *** p
Figure Legend Snippet: Neuron-specific depletion of Edis causes abnormal axonogenesis and neurodevelopment. (A) Confocal images of dorsal anterior regions of adult brains with neuron-specific expression of control shgfp (left panel) or shEdis (middle and right panels) driven by Elav-Gal4 driver. In control brain (left panel), anti-FasII antibody delineates the central complex (CC) as well as the vertical α and horizontal β and γ lobes of the mushroom bodies ( MB s), with γ lobes showing weaker FasII signal, as indicated. Depletion of Edis resulted in a spectrum of severe, age-dependent morphological defects in the MBs. In middle panel, one α lobe and one β lobe were mostly missing, and the remaining α and β lobes showed abnormal morphology. In right panel, both α lobes were largely missing, while the two β lobes crossed the midline and merged together. Scale bar: 20 μm. (B) Quantification of MB morphology phenotypes shown in A (Chi-squared test, sample numbers are shown on top, *** p

Techniques Used: Expressing

2) Product Images from "Characterization of enhancer fragments in Drosophila robo2"

Article Title: Characterization of enhancer fragments in Drosophila robo2

Journal: Fly

doi: 10.1080/19336934.2022.2126259

Lateral longitudinal axons labelled by GMR28F02 and GMR28G05 are distinct from FasII-positive lateral axon pathways.
Figure Legend Snippet: Lateral longitudinal axons labelled by GMR28F02 and GMR28G05 are distinct from FasII-positive lateral axon pathways.

Techniques Used:

3) Product Images from "Characterization of enhancer fragments in Drosophila robo2"

Article Title: Characterization of enhancer fragments in Drosophila robo2

Journal: bioRxiv

doi: 10.1101/2022.08.01.502399

GMR27F07 is expressed broadly in the embryonic ventral nerve cord. (A) GMR27F07/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed broadly throughout the brain and ventral nerve cord beginning around stage 11/12, and is also detectable in peripheral nervous system cells beginning around stage 13. (B-G) Ventral nerve cords from stage 12-17 GMR27F07/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed in pioneer neurons as early as stage 12 (B) and broad expression persists through stage 17 (G). GFP-positive longitudinal axons are visible in medial, intermediate, and lateral pathways (G, arrows; “m”, medial; “i”, intermediate; “l”, lateral).
Figure Legend Snippet: GMR27F07 is expressed broadly in the embryonic ventral nerve cord. (A) GMR27F07/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed broadly throughout the brain and ventral nerve cord beginning around stage 11/12, and is also detectable in peripheral nervous system cells beginning around stage 13. (B-G) Ventral nerve cords from stage 12-17 GMR27F07/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed in pioneer neurons as early as stage 12 (B) and broad expression persists through stage 17 (G). GFP-positive longitudinal axons are visible in medial, intermediate, and lateral pathways (G, arrows; “m”, medial; “i”, intermediate; “l”, lateral).

Techniques Used: Staining, Isolation, Expressing

GMR27H02 is expressed in a small number of longitudinal neurons in the embryonic ventral nerve cord. (A) GMR27H02/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed throughout the head/brain beginning around stage 12, and sparsely expressed in a few individually identifiable neurons per segment in the VNC, along with dorsal channel glia/TN exit glia. (B-G) Ventral nerve cords from stage 12-17 GMR27H02/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. The GFP-positive neurons are ipsilateral longitudinal neurons, with axons detectable in medial and intermediate axon pathways (F, arrows; “m”, medial; “i”, intermediate). Only two or three GFP-positive neurons are detectable in most abdominal VNC hemisegments.
Figure Legend Snippet: GMR27H02 is expressed in a small number of longitudinal neurons in the embryonic ventral nerve cord. (A) GMR27H02/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed throughout the head/brain beginning around stage 12, and sparsely expressed in a few individually identifiable neurons per segment in the VNC, along with dorsal channel glia/TN exit glia. (B-G) Ventral nerve cords from stage 12-17 GMR27H02/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. The GFP-positive neurons are ipsilateral longitudinal neurons, with axons detectable in medial and intermediate axon pathways (F, arrows; “m”, medial; “i”, intermediate). Only two or three GFP-positive neurons are detectable in most abdominal VNC hemisegments.

Techniques Used: Staining, Isolation

GMR28F12 lacks expression in the embryonic ventral nerve cord. (A) GMR28F12/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP expression is undetectable in the ventral nerve cord. GFP is strongly expressed in the visceral mesoderm (asterisks) and in ectodermal stripes (arrowheads) beginning around stage 13. Strong staining in some cells in brain/head region beginning around stage 13 and persisting through stage 17 in anterior head lobes. (B-G) Ventral nerve cords from stage 12-17 GMR28F12/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. There is little or no GFP expression in the ventral nerve cord at any of the examined developmental stages.
Figure Legend Snippet: GMR28F12 lacks expression in the embryonic ventral nerve cord. (A) GMR28F12/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP expression is undetectable in the ventral nerve cord. GFP is strongly expressed in the visceral mesoderm (asterisks) and in ectodermal stripes (arrowheads) beginning around stage 13. Strong staining in some cells in brain/head region beginning around stage 13 and persisting through stage 17 in anterior head lobes. (B-G) Ventral nerve cords from stage 12-17 GMR28F12/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. There is little or no GFP expression in the ventral nerve cord at any of the examined developmental stages.

Techniques Used: Expressing, Staining, Isolation

robo2 enhancer fragments and summary of GAL4 transgene expression patterns. (A) Schematic of the robo2/lea genomic region illustrating the intron/exon structure of the robo2 transcription unit (grey boxes, 5’ and 3’UTR; orange boxes, coding exons). Transcription of robo2 proceeds from right to left. Two robo2 transcripts with identical sequence are reported (robo2-RA and robo2-RB). Positions of putative enhancer fragments used to create GAL4 transgenic lines by Pfeiffer et al ( Pfeiffer et al. 2008 ) are shown below (“Putative Brain Enhancers”). Screenshot taken from http://flybase.org/cgi-bin/gbrowse2/dmel/?Search=1;name=FBgn0002543 . The location of the GAL4 enhancer trap transgene P{GawB}NP6273 (robo2 GAL4 ) is indicated by the cyan triangle. The insertion site is 217 bp upstream of the predicted robo2 transcriptional start site, and does not overlap with the GMR28C04 or GMR28B05 fragments. (B-R) GAL4 expression patterns in stage 16-17 ventral nerve cords from embryos carrying the indicated GAL4 transgenes along with UAS-TauMycGFP (UAS-TMG), labeled with antibodies against GFP (green; labels GAL4-expressing cells), FasII (red; labels a subset of longitudinal axon pathways), and HRP (blue; labels all axons). Lower panels show anti-GFP channel alone.
Figure Legend Snippet: robo2 enhancer fragments and summary of GAL4 transgene expression patterns. (A) Schematic of the robo2/lea genomic region illustrating the intron/exon structure of the robo2 transcription unit (grey boxes, 5’ and 3’UTR; orange boxes, coding exons). Transcription of robo2 proceeds from right to left. Two robo2 transcripts with identical sequence are reported (robo2-RA and robo2-RB). Positions of putative enhancer fragments used to create GAL4 transgenic lines by Pfeiffer et al ( Pfeiffer et al. 2008 ) are shown below (“Putative Brain Enhancers”). Screenshot taken from http://flybase.org/cgi-bin/gbrowse2/dmel/?Search=1;name=FBgn0002543 . The location of the GAL4 enhancer trap transgene P{GawB}NP6273 (robo2 GAL4 ) is indicated by the cyan triangle. The insertion site is 217 bp upstream of the predicted robo2 transcriptional start site, and does not overlap with the GMR28C04 or GMR28B05 fragments. (B-R) GAL4 expression patterns in stage 16-17 ventral nerve cords from embryos carrying the indicated GAL4 transgenes along with UAS-TauMycGFP (UAS-TMG), labeled with antibodies against GFP (green; labels GAL4-expressing cells), FasII (red; labels a subset of longitudinal axon pathways), and HRP (blue; labels all axons). Lower panels show anti-GFP channel alone.

Techniques Used: Expressing, Sequencing, Transgenic Assay, Labeling

GMR28D05 is expressed broadly in the embryonic ventral nerve cord. (A) GMR28D05/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed broadly throughout the brain and ventral nerve cord beginning around stage 11/12, and is also detectable in peripheral nervous system cells beginning around stage 13. Moderate GFP expression throughout the gut begins around stage 13 and persists through stage 17. (B-G) Ventral nerve cords from stage 12-17 GMR28D05/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed broadly in VNC neurons as early as stage 12 (B) and broad expression persists through stage 17 (G). Many GFP-positive axons are visible in both commissures and many longitudinal pathways.
Figure Legend Snippet: GMR28D05 is expressed broadly in the embryonic ventral nerve cord. (A) GMR28D05/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed broadly throughout the brain and ventral nerve cord beginning around stage 11/12, and is also detectable in peripheral nervous system cells beginning around stage 13. Moderate GFP expression throughout the gut begins around stage 13 and persists through stage 17. (B-G) Ventral nerve cords from stage 12-17 GMR28D05/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed broadly in VNC neurons as early as stage 12 (B) and broad expression persists through stage 17 (G). Many GFP-positive axons are visible in both commissures and many longitudinal pathways.

Techniques Used: Staining, Expressing, Isolation

GMR28G05 is expressed in a subset of commissural longitudinal neurons. (A) GMR28G05/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. Strong GFP expression in segmentally repeated clusters of neurons. GFP-positive cells wrapping external surface of nerve cord/motor nerve roots (likely peripheral glia) are prominent in mid-and late-stage embryos. (B-G) Ventral nerve cords from stage 12-17 GMR28G05/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. Lateral neuronal cell bodies are GFP-positive by stage 12 (B). Commissural axons visible crossing the midline in both commissures by stage 13 (C, arrows; “ac”, anterior commissure; “pc”, posterior commissure), and turn longitudinally at medial/intermediate positions by stage 14 (D, arrow). GFP-positive axons are detectable in lateral pathways at stages 16-17, at the same time or later than FasII-positive lateral axon pathways form (F,G, arrows).
Figure Legend Snippet: GMR28G05 is expressed in a subset of commissural longitudinal neurons. (A) GMR28G05/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. Strong GFP expression in segmentally repeated clusters of neurons. GFP-positive cells wrapping external surface of nerve cord/motor nerve roots (likely peripheral glia) are prominent in mid-and late-stage embryos. (B-G) Ventral nerve cords from stage 12-17 GMR28G05/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. Lateral neuronal cell bodies are GFP-positive by stage 12 (B). Commissural axons visible crossing the midline in both commissures by stage 13 (C, arrows; “ac”, anterior commissure; “pc”, posterior commissure), and turn longitudinally at medial/intermediate positions by stage 14 (D, arrow). GFP-positive axons are detectable in lateral pathways at stages 16-17, at the same time or later than FasII-positive lateral axon pathways form (F,G, arrows).

Techniques Used: Staining, Expressing, Isolation

GMR27D10 is stochastically expressed in a small number of longitudinal pioneer neurons in the ventral nerve cord. (A) GMR27D10/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed in a few midline-adjacent ipsilateral neurons per hemisegment at stage 12-13, with scattered expression in a few other neurons farther from the midline. GFP persists in these neurons and their axons through stage 17. GFP expression in brain/head begins as early as stage 11 and persists through stage 17. (B-G) Ventral nerve cords from stage 12-17 GMR27D10/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP-positive pioneer neurons are likely dMP2/vMP2 based on cell body position near the midline, axon projection pattern (C, arrows; one projects dorsally and one ventrally), and the location of their axons in the medial longitudinal pathway at late stages (F, arrow). GFP expression is stochastic in these neurons and either zero, one, or two of them express GFP in each hemisegment.
Figure Legend Snippet: GMR27D10 is stochastically expressed in a small number of longitudinal pioneer neurons in the ventral nerve cord. (A) GMR27D10/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed in a few midline-adjacent ipsilateral neurons per hemisegment at stage 12-13, with scattered expression in a few other neurons farther from the midline. GFP persists in these neurons and their axons through stage 17. GFP expression in brain/head begins as early as stage 11 and persists through stage 17. (B-G) Ventral nerve cords from stage 12-17 GMR27D10/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP-positive pioneer neurons are likely dMP2/vMP2 based on cell body position near the midline, axon projection pattern (C, arrows; one projects dorsally and one ventrally), and the location of their axons in the medial longitudinal pathway at late stages (F, arrow). GFP expression is stochastic in these neurons and either zero, one, or two of them express GFP in each hemisegment.

Techniques Used: Staining, Expressing, Isolation

GMR28C04 is expressed broadly in the embryonic ventral nerve cord. (A) GMR28C04/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed broadly throughout the brain and ventral nerve cord beginning around stage 11/12. The broad staining makes it difficult to discern specific aspects of GFP expression in the ventral nerve cords of whole embryos. GFP is also broadly expressed in body wall muscles (A, asterisk). (B-G) Ventral nerve cords from stage 12-17 GMR28C04/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed broadly in many neurons beginning at stage 13. GFP-positive commissural axons are visible at stage 13 (C, arrow; “c”, commissural) and GFP-positive longitudinal axons are detectable beginning at stage 14 (D, arrow) and persisting through stage 17 in medial and intermediate positions (G, arrows; “m”, medial; “i”, intermediate). The broad GFP expression in the VNC makes it difficult to link these longitudinal axons to specific neuronal cell bodies.
Figure Legend Snippet: GMR28C04 is expressed broadly in the embryonic ventral nerve cord. (A) GMR28C04/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed broadly throughout the brain and ventral nerve cord beginning around stage 11/12. The broad staining makes it difficult to discern specific aspects of GFP expression in the ventral nerve cords of whole embryos. GFP is also broadly expressed in body wall muscles (A, asterisk). (B-G) Ventral nerve cords from stage 12-17 GMR28C04/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed broadly in many neurons beginning at stage 13. GFP-positive commissural axons are visible at stage 13 (C, arrow; “c”, commissural) and GFP-positive longitudinal axons are detectable beginning at stage 14 (D, arrow) and persisting through stage 17 in medial and intermediate positions (G, arrows; “m”, medial; “i”, intermediate). The broad GFP expression in the VNC makes it difficult to link these longitudinal axons to specific neuronal cell bodies.

Techniques Used: Staining, Expressing, Isolation

GMR28F02 is expressed in a subset of commissural longitudinal neurons. (A) GMR28F02/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. Strong GFP expression in a segmentally repeated cluster of lateral neurons plus some midline-adjacent neurons. Peripheral GFP (possibly motor nerve or peripheral glia) detectable by stage 15. GFP detectable in visceral mesoderm from stage 12-13. Expressed in dorsal channel glia/TN exit glia beginning around stage 13. (B-G) Ventral nerve cords from stage 12-17 GMR28F02/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. Medial and lateral neuronal cell bodies are GFP-positive by stage 12 (B). Commissural axons visible crossing the midline in both commissures by stage 13 (C, arrows; “ac”, anterior commissure; “pc”, posterior commissure), and turn longitudinally at medial/intermediate and lateral positions by stage 14 (D, arrows; “m”, medial/intermediate; “l”, lateral). GFP-positive lateral pathway forms before FasII-positive axons are detectable in the lateral region (compare GFP and FasII staining in panel D).
Figure Legend Snippet: GMR28F02 is expressed in a subset of commissural longitudinal neurons. (A) GMR28F02/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. Strong GFP expression in a segmentally repeated cluster of lateral neurons plus some midline-adjacent neurons. Peripheral GFP (possibly motor nerve or peripheral glia) detectable by stage 15. GFP detectable in visceral mesoderm from stage 12-13. Expressed in dorsal channel glia/TN exit glia beginning around stage 13. (B-G) Ventral nerve cords from stage 12-17 GMR28F02/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. Medial and lateral neuronal cell bodies are GFP-positive by stage 12 (B). Commissural axons visible crossing the midline in both commissures by stage 13 (C, arrows; “ac”, anterior commissure; “pc”, posterior commissure), and turn longitudinally at medial/intermediate and lateral positions by stage 14 (D, arrows; “m”, medial/intermediate; “l”, lateral). GFP-positive lateral pathway forms before FasII-positive axons are detectable in the lateral region (compare GFP and FasII staining in panel D).

Techniques Used: Staining, Expressing, Isolation

GMR27D07 is expressed in commissural pioneer and longitudinal neurons in the ventral nerve cord. (A) GMR27D07/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed in clusters of neurons in the ventral nerve cord and throughout the brain/head region beginning by stages 12-13 and persisting through stage 17. Strong midline staining and distinct longitudinal pathways visible through stage 17. Peripheral GFP staining begins around stage 13. Strong GFP expression in lateral spots in the head that migrate anteriorly in later embryos (arrows). (B-G) Ventral nerve cords from stage 12-17 GMR27D07/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. Little or no GFP expression at stage 12; expression in small clusters of neurons begins stage 13, including some commissural pioneer neurons (C, arrow; “c”, commissural). Two distinct GFP-positive longitudinal pathways detectable by stage 14, persisting through stage 17 (G, arrows; “m”, medial; “i”, intermediate. Clusters of cells near the midline retain strong GFP expression through stage 17, with lower expression in a larger number of cells farther from the midline.
Figure Legend Snippet: GMR27D07 is expressed in commissural pioneer and longitudinal neurons in the ventral nerve cord. (A) GMR27D07/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed in clusters of neurons in the ventral nerve cord and throughout the brain/head region beginning by stages 12-13 and persisting through stage 17. Strong midline staining and distinct longitudinal pathways visible through stage 17. Peripheral GFP staining begins around stage 13. Strong GFP expression in lateral spots in the head that migrate anteriorly in later embryos (arrows). (B-G) Ventral nerve cords from stage 12-17 GMR27D07/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. Little or no GFP expression at stage 12; expression in small clusters of neurons begins stage 13, including some commissural pioneer neurons (C, arrow; “c”, commissural). Two distinct GFP-positive longitudinal pathways detectable by stage 14, persisting through stage 17 (G, arrows; “m”, medial; “i”, intermediate. Clusters of cells near the midline retain strong GFP expression through stage 17, with lower expression in a larger number of cells farther from the midline.

Techniques Used: Staining, Expressing, Isolation

GMR28A12 exhibits little or no expression in the embryonic ventral nerve cord. (A) GMR28A12/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. Little or no GFP expression is detectable in the ventral nerve cord, apart from a single bilateral pair of cells in each segment with processes extending laterally out of the CNS. GFP expressed in visceral mesoderm beginning stage 12 (arrowheads). GFP strongly expressed in salivary glands beginning around stage 14/15 (asterisks). Expression in anterior/head beginning stage 12 or earlier, persisting through stage 17 including anterior head lobes. (B-G) Ventral nerve cords from stage 12-17 GMR28A12/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. There is little or no GFP expression in the ventral nerve cord at any of the examined developmental stages. Cells near the midline with lateral processes are located dorsally to the ventral nerve cord, likely dorsal channel glia/TN exit glia (D, arrows).
Figure Legend Snippet: GMR28A12 exhibits little or no expression in the embryonic ventral nerve cord. (A) GMR28A12/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. Little or no GFP expression is detectable in the ventral nerve cord, apart from a single bilateral pair of cells in each segment with processes extending laterally out of the CNS. GFP expressed in visceral mesoderm beginning stage 12 (arrowheads). GFP strongly expressed in salivary glands beginning around stage 14/15 (asterisks). Expression in anterior/head beginning stage 12 or earlier, persisting through stage 17 including anterior head lobes. (B-G) Ventral nerve cords from stage 12-17 GMR28A12/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. There is little or no GFP expression in the ventral nerve cord at any of the examined developmental stages. Cells near the midline with lateral processes are located dorsally to the ventral nerve cord, likely dorsal channel glia/TN exit glia (D, arrows).

Techniques Used: Expressing, Staining, Isolation

GMR28A10 exhibits little or no expression in the embryonic ventral nerve cord. (A) GMR28A10/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. Little or no GFP expression is detectable in the ventral nerve cord. Scattered GFP expression in the anterior/head and in some peripheral cells at later stages. (B-G) Ventral nerve cords from stage 12-17 GMR28A10/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. There is little or no GFP expression in the ventral nerve cord at any of the examined developmental stages.
Figure Legend Snippet: GMR28A10 exhibits little or no expression in the embryonic ventral nerve cord. (A) GMR28A10/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. Little or no GFP expression is detectable in the ventral nerve cord. Scattered GFP expression in the anterior/head and in some peripheral cells at later stages. (B-G) Ventral nerve cords from stage 12-17 GMR28A10/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. There is little or no GFP expression in the ventral nerve cord at any of the examined developmental stages.

Techniques Used: Expressing, Staining, Isolation

GMR27G11 lacks expression in the embryonic ventral nerve cord. (A) GMR27G11/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. Little or no GFP expression is detectable in the ventral nerve cord. Sparse GFP expression is detectable in the anterior/head region in embryos from approximately stage 11 onwards. (B-G) Ventral nerve cords from stage 12-17 GMR27G11/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. There is little or no GFP expression in the ventral nerve cord at any of the examined developmental stages.
Figure Legend Snippet: GMR27G11 lacks expression in the embryonic ventral nerve cord. (A) GMR27G11/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. Little or no GFP expression is detectable in the ventral nerve cord. Sparse GFP expression is detectable in the anterior/head region in embryos from approximately stage 11 onwards. (B-G) Ventral nerve cords from stage 12-17 GMR27G11/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. There is little or no GFP expression in the ventral nerve cord at any of the examined developmental stages.

Techniques Used: Expressing, Staining, Isolation

GMR28E07 is expressed strongly in midline glia and pioneer longitudinal neurons. (A) GMR28E07/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is broadly expressed at a low level throughout the ventral nerve cord, and at a higher level in distinct cell clusters near the midline in each segment. GFP also expressed throughout the anterior/head and in body wall muscles and/or peripheral cells in the ectoderm. (B-G) Ventral nerve cords from stage 12-17 GMR28E07/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed in many neurons by stage 12, with little or no overlap with FasII (B). By stage 13 higher GFP expression is seen in midline cell clusters (C, arrowhead), with GFP-positive axons forming longitudinal tracts connecting adjacent segments (C, arrow; these tracts appear to be both GFP-positive and FasII-positive). GFP expression remains strong in midline cells (G, arrowhead) and longitudinal axons within medial and intermediate pathways (G, arrows; “m”, medial; “i”, intermediate) through stage 17. Expressed in dorsal channel glia/TN exit glia beginning around stage 13.
Figure Legend Snippet: GMR28E07 is expressed strongly in midline glia and pioneer longitudinal neurons. (A) GMR28E07/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is broadly expressed at a low level throughout the ventral nerve cord, and at a higher level in distinct cell clusters near the midline in each segment. GFP also expressed throughout the anterior/head and in body wall muscles and/or peripheral cells in the ectoderm. (B-G) Ventral nerve cords from stage 12-17 GMR28E07/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed in many neurons by stage 12, with little or no overlap with FasII (B). By stage 13 higher GFP expression is seen in midline cell clusters (C, arrowhead), with GFP-positive axons forming longitudinal tracts connecting adjacent segments (C, arrow; these tracts appear to be both GFP-positive and FasII-positive). GFP expression remains strong in midline cells (G, arrowhead) and longitudinal axons within medial and intermediate pathways (G, arrows; “m”, medial; “i”, intermediate) through stage 17. Expressed in dorsal channel glia/TN exit glia beginning around stage 13.

Techniques Used: Staining, Isolation, Expressing

GMR28E10 is expressed broadly in the embryonic ventral nerve cord. (A) GMR28E10/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed broadly throughout the epidermis, brain, and ventral nerve cord beginning around stage 11/12. The broad staining makes it difficult to discern specific aspects of GFP expression in the ventral nerve cords of whole embryos. (B-G) Ventral nerve cords from stage 12-17 GMR28E10/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed broadly in many neurons from stage 12 through stage 17. While some distinct GFP-positive commissural and longitudinal axons are discernable, the broad GFP expression throughout the nerve cord makes it difficult to identify other specific subsets of GFP-expressing neurons in these embryos.
Figure Legend Snippet: GMR28E10 is expressed broadly in the embryonic ventral nerve cord. (A) GMR28E10/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed broadly throughout the epidermis, brain, and ventral nerve cord beginning around stage 11/12. The broad staining makes it difficult to discern specific aspects of GFP expression in the ventral nerve cords of whole embryos. (B-G) Ventral nerve cords from stage 12-17 GMR28E10/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed broadly in many neurons from stage 12 through stage 17. While some distinct GFP-positive commissural and longitudinal axons are discernable, the broad GFP expression throughout the nerve cord makes it difficult to identify other specific subsets of GFP-expressing neurons in these embryos.

Techniques Used: Staining, Expressing, Isolation

GMR28B05 is expressed broadly in the embryonic ventral nerve cord. (A) GMR28B05/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed broadly throughout the epidermis. The broad staining makes it difficult to discern specific aspects of GFP expression in the ventral nerve cords of whole embryos. (B-G) Ventral nerve cords from stage 12-17 GMR28B05/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed broadly in many neurons and especially high in midline cells from stage 12 (B, arrowhead) through stage 17 (G, arrowhead). The broad GFP expression makes it difficult to identify specific subsets of GFP-expressing neurons in these embryos, although some GFP-positive longitudinal axons in medial and intermediate pathways are detectable.
Figure Legend Snippet: GMR28B05 is expressed broadly in the embryonic ventral nerve cord. (A) GMR28B05/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed broadly throughout the epidermis. The broad staining makes it difficult to discern specific aspects of GFP expression in the ventral nerve cords of whole embryos. (B-G) Ventral nerve cords from stage 12-17 GMR28B05/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed broadly in many neurons and especially high in midline cells from stage 12 (B, arrowhead) through stage 17 (G, arrowhead). The broad GFP expression makes it difficult to identify specific subsets of GFP-expressing neurons in these embryos, although some GFP-positive longitudinal axons in medial and intermediate pathways are detectable.

Techniques Used: Staining, Expressing, Isolation

GMR28D10 is expressed broadly in the embryonic ventral nerve cord. (A) GMR28D10/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed broadly throughout the epidermis, brain, and ventral nerve cord beginning around stage 11/12. The broad staining makes it difficult to discern specific aspects of GFP expression in the ventral nerve cords of whole embryos. (B-G) Ventral nerve cords from stage 12-17 GMR28D10/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed broadly in many neurons from stage 12 through stage 17. While some distinct GFP-positive commissural and longitudinal axons are discernable, the broad GFP expression throughout the nerve cord makes it difficult to identify other specific subsets of GFP-expressing neurons in these embryos.
Figure Legend Snippet: GMR28D10 is expressed broadly in the embryonic ventral nerve cord. (A) GMR28D10/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed broadly throughout the epidermis, brain, and ventral nerve cord beginning around stage 11/12. The broad staining makes it difficult to discern specific aspects of GFP expression in the ventral nerve cords of whole embryos. (B-G) Ventral nerve cords from stage 12-17 GMR28D10/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed broadly in many neurons from stage 12 through stage 17. While some distinct GFP-positive commissural and longitudinal axons are discernable, the broad GFP expression throughout the nerve cord makes it difficult to identify other specific subsets of GFP-expressing neurons in these embryos.

Techniques Used: Staining, Expressing, Isolation

Expression of a robo2 GAL4 enhancer trap allele (A) robo2 GAL4 /UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is broadly expressed throughout the epidermis in these embryos throughout embryonic development. GFP expression in midline cells of the ventral nerve cord is detectable (arrowhead) but other aspects of internal GFP expression are obscured by the strong epidermal staining. (B-G) Ventral nerve cords from stage 12-17 robo2 GAL4 /UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is strongly expressed in midline cells from stage 12-13 through stage 17 (C,G, arrowhead), and is detectable on early pioneer axons (C, arrow) that remain detectable in medial and intermediate longitudinal pathways in older embryos (G, arrow). As in the whole mount embryos in (A), the broad GFP expression throughout the nerve cord makes it difficult to identify other specific subsets of GFP-expressing neurons in these embryos.
Figure Legend Snippet: Expression of a robo2 GAL4 enhancer trap allele (A) robo2 GAL4 /UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is broadly expressed throughout the epidermis in these embryos throughout embryonic development. GFP expression in midline cells of the ventral nerve cord is detectable (arrowhead) but other aspects of internal GFP expression are obscured by the strong epidermal staining. (B-G) Ventral nerve cords from stage 12-17 robo2 GAL4 /UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is strongly expressed in midline cells from stage 12-13 through stage 17 (C,G, arrowhead), and is detectable on early pioneer axons (C, arrow) that remain detectable in medial and intermediate longitudinal pathways in older embryos (G, arrow). As in the whole mount embryos in (A), the broad GFP expression throughout the nerve cord makes it difficult to identify other specific subsets of GFP-expressing neurons in these embryos.

Techniques Used: Expressing, Staining, Isolation

Lateral longitudinal axons labeled by GMR28F02 and GMR28G05 are distinct from FasII-positive lateral axon pathways. (A) Stage 16 robo2 myc-robo2 embryo stained with anti-myc (green), anti-FasII (red) and anti-HRP (blue) antibodies. Individual channels are shown in grayscale. Lower panel shows confocal x,z cross-sections through the area outlined by the white box. (B,C) Stage 16 GMR28F02/UAS-TMG (B) and GMR28G05/UAS-TMG (C) embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower panels show confocal x,z cross-sections through the area outlined by the white box. (A’-C’) Enlarged view of the confocal x,z cross-sections from panels (A-C). Arrows indicate the position of the dorsal lateral FasII axon pathway, and arrowheads indicate the position of myc-positive (A’) or GFP-positive (B’,C’) axons. While myc-positive axons in A’ overlap with FasII-positive lateral axon pathways, GFP-positive axons in B’ and C’ do not overlap with FasII, indicating that GMR28F02 and GMR28G05 label lateral longitudinal axons that are not part of FasII-positive pathways.
Figure Legend Snippet: Lateral longitudinal axons labeled by GMR28F02 and GMR28G05 are distinct from FasII-positive lateral axon pathways. (A) Stage 16 robo2 myc-robo2 embryo stained with anti-myc (green), anti-FasII (red) and anti-HRP (blue) antibodies. Individual channels are shown in grayscale. Lower panel shows confocal x,z cross-sections through the area outlined by the white box. (B,C) Stage 16 GMR28F02/UAS-TMG (B) and GMR28G05/UAS-TMG (C) embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower panels show confocal x,z cross-sections through the area outlined by the white box. (A’-C’) Enlarged view of the confocal x,z cross-sections from panels (A-C). Arrows indicate the position of the dorsal lateral FasII axon pathway, and arrowheads indicate the position of myc-positive (A’) or GFP-positive (B’,C’) axons. While myc-positive axons in A’ overlap with FasII-positive lateral axon pathways, GFP-positive axons in B’ and C’ do not overlap with FasII, indicating that GMR28F02 and GMR28G05 label lateral longitudinal axons that are not part of FasII-positive pathways.

Techniques Used: Labeling, Staining

GMR28D12 is expressed broadly in the embryonic ventral nerve cord. (A) GMR28D12/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed broadly throughout the epidermis. The broad staining makes it difficult to discern specific aspects of GFP expression in the ventral nerve cords of whole embryos. (B-G) Ventral nerve cords from stage 12-17 GMR28D12/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed broadly in many neurons and elevated in midline cel ls from stage 12 (B, arrowhead) through stage 17 (G, arrowhead). The broad GFP expression makes it difficult to identify specific subsets of GFP-expressing neurons in these embryos.
Figure Legend Snippet: GMR28D12 is expressed broadly in the embryonic ventral nerve cord. (A) GMR28D12/UAS-TMG embryos stained with an anti-GFP antibody (brown). Representative embryos from various developmental stages are arranged in order of increasing age from left to right. Lateral view of youngest embryo on left; all others ventral side up. GFP is expressed broadly throughout the epidermis. The broad staining makes it difficult to discern specific aspects of GFP expression in the ventral nerve cords of whole embryos. (B-G) Ventral nerve cords from stage 12-17 GMR28D12/UAS-TMG embryos stained with anti-GFP (green), anti-FasII (red) and anti-HRP (blue) antibodies. Lower images show isolated channels for each of the three antibodies. GFP is expressed broadly in many neurons and elevated in midline cel ls from stage 12 (B, arrowhead) through stage 17 (G, arrowhead). The broad GFP expression makes it difficult to identify specific subsets of GFP-expressing neurons in these embryos.

Techniques Used: Staining, Expressing, Isolation

4) Product Images from "Molecular Dissection of DAAM Function during Axon Growth in Drosophila Embryonic Neurons"

Article Title: Molecular Dissection of DAAM Function during Axon Growth in Drosophila Embryonic Neurons

Journal: Cells

doi: 10.3390/cells11091487

The FH2R/K-A mutant form of DAAM fails to rescue the MB axonal defects observed in DAAM Ex4 . ( A ) Schematic representation of the organization of the mushroom bodies. ( B – F′ ) Confocal images of MBs stained for FasII (green) to label the α and β-lobes; with the exception of wild-type; ( B ) two representative examples are shown for each genotype. The DAAM Ex4 mutant ( C , C′ ) MBs exhibit various defects in axonal development resulting in missing, shorter, thinner or thicker lobes, which can be perfectly rescued by UAS-DAAM expression ( D , D′ ). As compared to this, the expression of UAS-DAAM FH2R/K-A , in addition to various effects on the dorsal lobes, results in overprojection of the β axons ( E , E′ ) which is very similar to the effect of DAD-G expression ( F , F′ ). ( G ) Quantification of the axonal growth and guidance and defects detected in the MB α-lobes of adults of the genotypes indicated. ( H ) Quantification of the axonal growth and guidance and defects detected in the MB β-lobes of adults of the genotypes indicated. ( I ) Quantification of the β-lobe fusion phenotype in the MBs of adults of the genotypes indicated. Chi-square or Fisher’s exact tests were used for statistical analysis. * p
Figure Legend Snippet: The FH2R/K-A mutant form of DAAM fails to rescue the MB axonal defects observed in DAAM Ex4 . ( A ) Schematic representation of the organization of the mushroom bodies. ( B – F′ ) Confocal images of MBs stained for FasII (green) to label the α and β-lobes; with the exception of wild-type; ( B ) two representative examples are shown for each genotype. The DAAM Ex4 mutant ( C , C′ ) MBs exhibit various defects in axonal development resulting in missing, shorter, thinner or thicker lobes, which can be perfectly rescued by UAS-DAAM expression ( D , D′ ). As compared to this, the expression of UAS-DAAM FH2R/K-A , in addition to various effects on the dorsal lobes, results in overprojection of the β axons ( E , E′ ) which is very similar to the effect of DAD-G expression ( F , F′ ). ( G ) Quantification of the axonal growth and guidance and defects detected in the MB α-lobes of adults of the genotypes indicated. ( H ) Quantification of the axonal growth and guidance and defects detected in the MB β-lobes of adults of the genotypes indicated. ( I ) Quantification of the β-lobe fusion phenotype in the MBs of adults of the genotypes indicated. Chi-square or Fisher’s exact tests were used for statistical analysis. * p

Techniques Used: Mutagenesis, Staining, Expressing

The effect of CDAAM overexpression on neuronal morphology. ( A – D ) Morphology of the FasII-positive motoraxons in the ventral nerve cord of control ( Elav-Gal4 ) ( A ), CDAAM ( B ), CDAAM I732A ( C ) and CDAAM FH2R/K-A ( D ) expressing Drosophila embryos. Scale bar represents 50 µm. ( E – G″ ) Representative images of axonal and growth cone morphology of primary neurons derived from control ( E – E″ ), CDAAM ( F – F″ ) or CDAAM I732A ( G – G″ ) expressing Drosophila embryos. The actin cytoskeleton was labelled by phalloidin (green), and microtubules were detected by an anti-tubulin antibody (magenta). Scale bar represents 5 µm. ( H ) Statistical analysis of the length of axonal microtubule bundles. Scatter plots show the values of the individual cells and the means of the independent experiments. ANOVA was used for statistical analysis. Tukey’s post hoc test was used for multiple comparison. ** p
Figure Legend Snippet: The effect of CDAAM overexpression on neuronal morphology. ( A – D ) Morphology of the FasII-positive motoraxons in the ventral nerve cord of control ( Elav-Gal4 ) ( A ), CDAAM ( B ), CDAAM I732A ( C ) and CDAAM FH2R/K-A ( D ) expressing Drosophila embryos. Scale bar represents 50 µm. ( E – G″ ) Representative images of axonal and growth cone morphology of primary neurons derived from control ( E – E″ ), CDAAM ( F – F″ ) or CDAAM I732A ( G – G″ ) expressing Drosophila embryos. The actin cytoskeleton was labelled by phalloidin (green), and microtubules were detected by an anti-tubulin antibody (magenta). Scale bar represents 5 µm. ( H ) Statistical analysis of the length of axonal microtubule bundles. Scatter plots show the values of the individual cells and the means of the independent experiments. ANOVA was used for statistical analysis. Tukey’s post hoc test was used for multiple comparison. ** p

Techniques Used: Over Expression, Expressing, Derivative Assay

Morphological analysis of primary neurons derived from DAAM Ex4 mutant Drosophila embryos expressing DAAM FH2R/K-A . ( A – B″ ) Representative images of the axonal and growth cone morphology of primary neurons derived from DAAM Ex4 ;Elav-Gal4 control ( A – A″ ) and DAAM FH2R/K-A ( B – B″ ) expressing Drosophila embryos. The actin cytoskeleton was labelled by phalloidin (green), and microtubules were detected by an anti-tubulin antibody (magenta). Scale bar represents 5 µm. ( C ) Statistical analysis of the length of axonal microtubule bundles. Scatter plots show the values of the individual cells and the means of the independent experiments. ( D ) Scatter plots show the frequency of microtubule morphologies in the growth cone. ( E ) Statistical analysis of the axonal filopodia numbers. Scatter plots represent the values of the individual cells and the means of the independent experiments. ( F ) Scatter plots show the frequency of growth cone morphologies labeled by phalloidin. ANOVA was used for statistical analysis. Tukey’s post hoc test was used for multiple comparison. ** p
Figure Legend Snippet: Morphological analysis of primary neurons derived from DAAM Ex4 mutant Drosophila embryos expressing DAAM FH2R/K-A . ( A – B″ ) Representative images of the axonal and growth cone morphology of primary neurons derived from DAAM Ex4 ;Elav-Gal4 control ( A – A″ ) and DAAM FH2R/K-A ( B – B″ ) expressing Drosophila embryos. The actin cytoskeleton was labelled by phalloidin (green), and microtubules were detected by an anti-tubulin antibody (magenta). Scale bar represents 5 µm. ( C ) Statistical analysis of the length of axonal microtubule bundles. Scatter plots show the values of the individual cells and the means of the independent experiments. ( D ) Scatter plots show the frequency of microtubule morphologies in the growth cone. ( E ) Statistical analysis of the axonal filopodia numbers. Scatter plots represent the values of the individual cells and the means of the independent experiments. ( F ) Scatter plots show the frequency of growth cone morphologies labeled by phalloidin. ANOVA was used for statistical analysis. Tukey’s post hoc test was used for multiple comparison. ** p

Techniques Used: Derivative Assay, Mutagenesis, Expressing, Labeling

Biochemical characterization of the FH2R/K-A mutant form of DAAM. ( A – B′ ) Ribbon diagram ( A , B ) and surface charge distribution ( A′ , B′ ) of the wild-type and R/K-A mutant FH2 domain of Drosophila DAAM. The region affected by the R/K-A is encircled in red (dashed oval in A′ , B′ ). ( C ) A GST-pull down assay, carried out by using GST-tagged wild-type and R/K-A mutant FH2 recombinant proteins in combination with purified tubulin. Eluates of the GST-pull down were analyzed by gel electrophoresis and Western blot. Recombinant proteins were visualized by Coomassie blue staining and bound tubulin was detected by an anti-tubulin antibody. Note the presence of tubulin in samples with GST::FH2, and the lack of tubulin when GST::FH2R/K-A was immobilized on the beads. ( D ) Western blot analysis of samples obtained from a microtubule co-sedimentation assay. Tubulin was detected by an anti-tubulin antibody, while the Flag-tagged wild-type and R/K-A mutant form of DAAM FH2 was detected by an anti-Flag antibody. Flag-tagged FH2 protein was detected in the pellet. In contrast, the 3xFLAG::FH2-R/K-A did not co-sediment with microtubules. I—input, SN—supernatant, P—pellet.
Figure Legend Snippet: Biochemical characterization of the FH2R/K-A mutant form of DAAM. ( A – B′ ) Ribbon diagram ( A , B ) and surface charge distribution ( A′ , B′ ) of the wild-type and R/K-A mutant FH2 domain of Drosophila DAAM. The region affected by the R/K-A is encircled in red (dashed oval in A′ , B′ ). ( C ) A GST-pull down assay, carried out by using GST-tagged wild-type and R/K-A mutant FH2 recombinant proteins in combination with purified tubulin. Eluates of the GST-pull down were analyzed by gel electrophoresis and Western blot. Recombinant proteins were visualized by Coomassie blue staining and bound tubulin was detected by an anti-tubulin antibody. Note the presence of tubulin in samples with GST::FH2, and the lack of tubulin when GST::FH2R/K-A was immobilized on the beads. ( D ) Western blot analysis of samples obtained from a microtubule co-sedimentation assay. Tubulin was detected by an anti-tubulin antibody, while the Flag-tagged wild-type and R/K-A mutant form of DAAM FH2 was detected by an anti-Flag antibody. Flag-tagged FH2 protein was detected in the pellet. In contrast, the 3xFLAG::FH2-R/K-A did not co-sediment with microtubules. I—input, SN—supernatant, P—pellet.

Techniques Used: Mutagenesis, Pull Down Assay, Recombinant, Purification, Nucleic Acid Electrophoresis, Western Blot, Staining, Sedimentation

Morphological analysis of S2 cells expressing GFP-tagged wild-type and mutant forms of CDAAM. ( A – D″ ) Representative images of the cytoskeletal organization of control ( A – A″ , a , a′ ), CDAAM ( B – B″ , b , b′ ), CDAAMI732A ( C – C″ , c , c′ ) and CDAAMFH2R/K-A ( D – D″ , d , d′ ) expressing S2 cells. GFP or GFP-tagged CDAAM was detected by an anti-GFP antibody (green), and microtubules are visualized by an anti-tubulin antibody (magenta). Filamentous organization of CDAAM was visible in the cortical lamellipodial region of CDAAM and CDAAMI732A expressing cells (see the insets in b , b′ , c , c′, respectively), which is not present in control cells ( a , a′ ), and in cells expressing CDAAMFH2R/K-A ( d , d′ ) where most of the GFP signal accumulates into cytoplasmic foci (arrows in D″ ). Scale bar represents 5 µm.
Figure Legend Snippet: Morphological analysis of S2 cells expressing GFP-tagged wild-type and mutant forms of CDAAM. ( A – D″ ) Representative images of the cytoskeletal organization of control ( A – A″ , a , a′ ), CDAAM ( B – B″ , b , b′ ), CDAAMI732A ( C – C″ , c , c′ ) and CDAAMFH2R/K-A ( D – D″ , d , d′ ) expressing S2 cells. GFP or GFP-tagged CDAAM was detected by an anti-GFP antibody (green), and microtubules are visualized by an anti-tubulin antibody (magenta). Filamentous organization of CDAAM was visible in the cortical lamellipodial region of CDAAM and CDAAMI732A expressing cells (see the insets in b , b′ , c , c′, respectively), which is not present in control cells ( a , a′ ), and in cells expressing CDAAMFH2R/K-A ( d , d′ ) where most of the GFP signal accumulates into cytoplasmic foci (arrows in D″ ). Scale bar represents 5 µm.

Techniques Used: Expressing, Mutagenesis

Morphological analysis of the nervous system of DAAM and frl single- and double-mutant embryos. A) Morphology of FasII-positive motoraxons in the ventral nerve cord of wild-type ( A ), frl59 ( A′ ), DAAM Ex4 ( A″ ) and DAAM Ex4 /frl 59 ( A‴ ) embryos. Scale bar represents 50 µm. ( B ) Frequency of ISNb phenotypes in wild-type and mutant embryos. ( C – C′ ) Schematic representation of wild-type and mutant (stalled) motoraxons (TN—transverse nerve, ISNb—intersegmental nerve b, ISNa—intersegmental nerve a, SNa—segmental nerve a). ( D ) Morphology of the FasII-positive motoraxons in the ISNb of wild-type ( D ), frl 59 ( D′ ), DAAM Ex4 ( D″ ) and DAAM Ex4 ; frl 59 ( D‴ ) embryos. Arrows in ( D″ ) and ( D‴ ) point to the stalled ISNb. Scale bar represents 10 µm.
Figure Legend Snippet: Morphological analysis of the nervous system of DAAM and frl single- and double-mutant embryos. A) Morphology of FasII-positive motoraxons in the ventral nerve cord of wild-type ( A ), frl59 ( A′ ), DAAM Ex4 ( A″ ) and DAAM Ex4 /frl 59 ( A‴ ) embryos. Scale bar represents 50 µm. ( B ) Frequency of ISNb phenotypes in wild-type and mutant embryos. ( C – C′ ) Schematic representation of wild-type and mutant (stalled) motoraxons (TN—transverse nerve, ISNb—intersegmental nerve b, ISNa—intersegmental nerve a, SNa—segmental nerve a). ( D ) Morphology of the FasII-positive motoraxons in the ISNb of wild-type ( D ), frl 59 ( D′ ), DAAM Ex4 ( D″ ) and DAAM Ex4 ; frl 59 ( D‴ ) embryos. Arrows in ( D″ ) and ( D‴ ) point to the stalled ISNb. Scale bar represents 10 µm.

Techniques Used: Mutagenesis

The actin-processing activity of DAAM plays a role in axon development in primary neurons. ( A – D″ ) Representative images of the axonal and growth cone morphology of primary neurons derived from DAAM Ex4 ;Elav-Gal4 control ( A – A″ ) and transgene expressing ( B – B″ - DAAM ; C – C″ - DAAM I732A ; D – D″ - DAAM K881A ) Drosophila embryos. The actin cytoskeleton was labelled by phalloidin (green), and microtubules were detected by an anti-tubulin antibody (magenta). Scale bar represents 5 µm. ( E ) Statistical analysis of the length of the axonal microtubule bundles. Scatter plots show the values of the individual cells and the means of the independent experiments. ( F ) Scatter plots show the frequency of microtubule morphologies in the growth cone. ( G ) Statistical analysis of the axonal filopodia numbers. Scatter plots represent the values of the individual cells and the means of the independent experiments. ( H ) Scatter plots show the frequency of growth cone morphologies labelled by phalloidin. ANOVA was used for statistical analysis. Tukey’s post hoc test was used for multiple comparison. * p
Figure Legend Snippet: The actin-processing activity of DAAM plays a role in axon development in primary neurons. ( A – D″ ) Representative images of the axonal and growth cone morphology of primary neurons derived from DAAM Ex4 ;Elav-Gal4 control ( A – A″ ) and transgene expressing ( B – B″ - DAAM ; C – C″ - DAAM I732A ; D – D″ - DAAM K881A ) Drosophila embryos. The actin cytoskeleton was labelled by phalloidin (green), and microtubules were detected by an anti-tubulin antibody (magenta). Scale bar represents 5 µm. ( E ) Statistical analysis of the length of the axonal microtubule bundles. Scatter plots show the values of the individual cells and the means of the independent experiments. ( F ) Scatter plots show the frequency of microtubule morphologies in the growth cone. ( G ) Statistical analysis of the axonal filopodia numbers. Scatter plots represent the values of the individual cells and the means of the independent experiments. ( H ) Scatter plots show the frequency of growth cone morphologies labelled by phalloidin. ANOVA was used for statistical analysis. Tukey’s post hoc test was used for multiple comparison. * p

Techniques Used: Activity Assay, Derivative Assay, Expressing

Morphological analysis of primary neurons derived from DAAM and frl single- and double-mutant embryos. ( A – D″ ) Representative images of primary neurons derived from wild-type ( A – A″ ), DAAM Ex4 ( B – B″ ), frl 59 ( C – C″ ) and DAAM Ex4 ; frl 59 ( D – D″ ) embryos. The actin cytoskeleton was labelled by phalloidin (green); microtubules were visualized by an anti-tubulin (magenta) antibody. Scale bar represents 5 µm. ( E – F″ ) Examples for cytoskeleton organization of axonal growth cones labelled by phalloidin (green) and anti-tubulin antibody (magenta). Scale bar represents 5 µm. ( G ) Statistical analysis of axonal length of axonal microtubule bundles. Scatter plots show the values of the individual cells and the means of the independent experiments. ( H ) Scatter plots show the frequency of microtubule morphologies in the growth cone. ( I ) Statistical analysis of the axonal filopodia numbers. Scatter plots represent the values of the individual cells and the means of the independent experiments. ( J ) Scatter plots show the frequency of growth cone morphologies labelled by phalloidin. ANOVA was used for statistical analysis. Tukey’s post hoc test was used for multiple comparison. * p
Figure Legend Snippet: Morphological analysis of primary neurons derived from DAAM and frl single- and double-mutant embryos. ( A – D″ ) Representative images of primary neurons derived from wild-type ( A – A″ ), DAAM Ex4 ( B – B″ ), frl 59 ( C – C″ ) and DAAM Ex4 ; frl 59 ( D – D″ ) embryos. The actin cytoskeleton was labelled by phalloidin (green); microtubules were visualized by an anti-tubulin (magenta) antibody. Scale bar represents 5 µm. ( E – F″ ) Examples for cytoskeleton organization of axonal growth cones labelled by phalloidin (green) and anti-tubulin antibody (magenta). Scale bar represents 5 µm. ( G ) Statistical analysis of axonal length of axonal microtubule bundles. Scatter plots show the values of the individual cells and the means of the independent experiments. ( H ) Scatter plots show the frequency of microtubule morphologies in the growth cone. ( I ) Statistical analysis of the axonal filopodia numbers. Scatter plots represent the values of the individual cells and the means of the independent experiments. ( J ) Scatter plots show the frequency of growth cone morphologies labelled by phalloidin. ANOVA was used for statistical analysis. Tukey’s post hoc test was used for multiple comparison. * p

Techniques Used: Derivative Assay, Mutagenesis

5) Product Images from "Analysis of growth cone extension in standardized coordinates highlights self-organization rules during wiring of the Drosophila visual system"

Article Title: Analysis of growth cone extension in standardized coordinates highlights self-organization rules during wiring of the Drosophila visual system

Journal: PLoS Genetics

doi: 10.1371/journal.pgen.1009857

Changes in R3 or R4 cell identity lead to changes in final targeting. (A) Schematic of NSP wiring topology. Solid circles: Landing points of R1-R6 at the lamina (“heels”); open circles: target locations of R1-R6 growth cones. T3’: target of fate-transformed R3s; T0: target located within the bundle of interest (though targeted by R cells from other bundles in NSP wiring). R1-R6 are color coded consistently in all schematics. (B-C) Representative schematics and images of bundles from (B) sev > Fz (38 hrs APF) and (C) sev > N ic (45 hrs APF) flies (see S1B and S1C Fig for more examples). Top panels: schematics of wild-type or altered wiring topology. Bottom panels: confocal images of representative bundles. Photoreceptor growth cones were segmented, pseudo-colored, and intensity scaled for visualization ( Materials and Methods ). Red: sev > RFP expression; green: mδ0 . 5-GFP expression; white: Fasciclin 2 (FasII) antibody staining. White ellipses: targets. Scale bar: 5 μm.
Figure Legend Snippet: Changes in R3 or R4 cell identity lead to changes in final targeting. (A) Schematic of NSP wiring topology. Solid circles: Landing points of R1-R6 at the lamina (“heels”); open circles: target locations of R1-R6 growth cones. T3’: target of fate-transformed R3s; T0: target located within the bundle of interest (though targeted by R cells from other bundles in NSP wiring). R1-R6 are color coded consistently in all schematics. (B-C) Representative schematics and images of bundles from (B) sev > Fz (38 hrs APF) and (C) sev > N ic (45 hrs APF) flies (see S1B and S1C Fig for more examples). Top panels: schematics of wild-type or altered wiring topology. Bottom panels: confocal images of representative bundles. Photoreceptor growth cones were segmented, pseudo-colored, and intensity scaled for visualization ( Materials and Methods ). Red: sev > RFP expression; green: mδ0 . 5-GFP expression; white: Fasciclin 2 (FasII) antibody staining. White ellipses: targets. Scale bar: 5 μm.

Techniques Used: Transformation Assay, Expressing, Staining

6) Product Images from "On the role of photoreceptor identity in controlling accurate wiring of the Drosophila visual circuit"

Article Title: On the role of photoreceptor identity in controlling accurate wiring of the Drosophila visual circuit

Journal: bioRxiv

doi: 10.1101/2020.10.13.337865

Final targeting of sev > Fz and sev > N ic flies. (A-B) Schematics (top panels) and confocal images (bottom panels) of bundles from three (A) sev > Fz (38 hrs APF) and (B) sev > N ic (45 hrs APF) flies. Top panels: schematic of wild-type or altered wiring topology. Solid or open circles: starting points (‘heels”) or targets (respectively); colors coordinated between R cells and targets. T3’: target of fate-altered R3s; T0: target located within the bundle of interest (though targeted by R cells from other bundles in NSP wiring). Bottom panels: confocal images of representative bundles. Photoreceptor growth cones are segmented and pseudo-colored (Methods) and intensity scaled for visualization. Red: sev > RFP expression; green: mδ-GFP expression; white: Fasciclin 2 (FasII) antibody staining. White ellipses: targets. Scale bar: 5 μm.
Figure Legend Snippet: Final targeting of sev > Fz and sev > N ic flies. (A-B) Schematics (top panels) and confocal images (bottom panels) of bundles from three (A) sev > Fz (38 hrs APF) and (B) sev > N ic (45 hrs APF) flies. Top panels: schematic of wild-type or altered wiring topology. Solid or open circles: starting points (‘heels”) or targets (respectively); colors coordinated between R cells and targets. T3’: target of fate-altered R3s; T0: target located within the bundle of interest (though targeted by R cells from other bundles in NSP wiring). Bottom panels: confocal images of representative bundles. Photoreceptor growth cones are segmented and pseudo-colored (Methods) and intensity scaled for visualization. Red: sev > RFP expression; green: mδ-GFP expression; white: Fasciclin 2 (FasII) antibody staining. White ellipses: targets. Scale bar: 5 μm.

Techniques Used: Expressing, Staining

Changes in R3 or R4 cell identity lead to changes in final targeting. (A-B) Representative schematics and images of bundles from (A) sev > Fz (38 hrs APF) and (B) sev > N ic (45 hrs APF) flies (see Figure 1 – figure supplement 2 for more examples). Top panels: schematic of wild-type or altered wiring topology. Solid or open circles: starting points (‘heels”) or targets (respectively); colors coordinated between R cells and targets. T3’: target of fate-altered R3s; T0: target located within the bundle of interest (though targeted by R cells from other bundles in NSP wiring). Bottom panels: confocal images of representative bundles. Photoreceptor growth cones were segmented, pseudo-colored, and intensity scaled for visualization (Methods). Red: sev > RFP expression; green: mδ-GFP expression; white: Fasciclin 2 (FasII) antibody staining. White ellipses: targets. Scale bar: 5 μm.
Figure Legend Snippet: Changes in R3 or R4 cell identity lead to changes in final targeting. (A-B) Representative schematics and images of bundles from (A) sev > Fz (38 hrs APF) and (B) sev > N ic (45 hrs APF) flies (see Figure 1 – figure supplement 2 for more examples). Top panels: schematic of wild-type or altered wiring topology. Solid or open circles: starting points (‘heels”) or targets (respectively); colors coordinated between R cells and targets. T3’: target of fate-altered R3s; T0: target located within the bundle of interest (though targeted by R cells from other bundles in NSP wiring). Bottom panels: confocal images of representative bundles. Photoreceptor growth cones were segmented, pseudo-colored, and intensity scaled for visualization (Methods). Red: sev > RFP expression; green: mδ-GFP expression; white: Fasciclin 2 (FasII) antibody staining. White ellipses: targets. Scale bar: 5 μm.

Techniques Used: Expressing, Staining

7) Product Images from "Steroid Hormone Entry into the Brain Requires a Membrane Transporter in Drosophila"

Article Title: Steroid Hormone Entry into the Brain Requires a Membrane Transporter in Drosophila

Journal: Current biology : CB

doi: 10.1016/j.cub.2019.11.085

EcI in the BBB Is Required for Ecdysone-Mediated Neuronal Events during Development. (A) Developmental changes of the CNS morphology in control, EcR RNAi, and EcI RNAi animals. 9–137-Gal4 > UAS-dicer2 was used to induce RNAi in the BBB. Nuclei in the CNS were stained with Hoechst 33342. Representative images of the CNS at different stages were combined into a single panel. White arrow indicates the failure of the separation of the subesophageal and thoracic ganglia in the CNS of EcR RNAi animals. Consistent phenotypes were observed in all samples examined (n = 10–20 CNS samples per genotype). Scale bar, 200 μm. (B, C, D) Ecdysone-mediated neuronal differentiation (B), remodeling (C) and apoptosis (D) during development. 9–137-Gal4 > UAS-dicer2 ] and indicated by white and yellow brackets, respectively. (C) CNS Immunostaining at 24 hAPF for Fasciclin-II (Fas-II) to label mushroom body lobes. α and β indicate newly extending axonal lobes of α/β neurons, and γ indicates larval-specific axonal lobe of γ neurons. Consistent phenotypes were observed in all samples examined (n = 18–22 CNS samples per genotype). (D) CNS Immunostaining at 0 and 24 hAPF for corazonin. Upper panels show dorso-lateral corazonin-producing neurons (dlCrz) and lower panels show ventral nerve cord corazonin-producing neurons (vCrz). for quantitative analyses of (B) and (D).
Figure Legend Snippet: EcI in the BBB Is Required for Ecdysone-Mediated Neuronal Events during Development. (A) Developmental changes of the CNS morphology in control, EcR RNAi, and EcI RNAi animals. 9–137-Gal4 > UAS-dicer2 was used to induce RNAi in the BBB. Nuclei in the CNS were stained with Hoechst 33342. Representative images of the CNS at different stages were combined into a single panel. White arrow indicates the failure of the separation of the subesophageal and thoracic ganglia in the CNS of EcR RNAi animals. Consistent phenotypes were observed in all samples examined (n = 10–20 CNS samples per genotype). Scale bar, 200 μm. (B, C, D) Ecdysone-mediated neuronal differentiation (B), remodeling (C) and apoptosis (D) during development. 9–137-Gal4 > UAS-dicer2 ] and indicated by white and yellow brackets, respectively. (C) CNS Immunostaining at 24 hAPF for Fasciclin-II (Fas-II) to label mushroom body lobes. α and β indicate newly extending axonal lobes of α/β neurons, and γ indicates larval-specific axonal lobe of γ neurons. Consistent phenotypes were observed in all samples examined (n = 18–22 CNS samples per genotype). (D) CNS Immunostaining at 0 and 24 hAPF for corazonin. Upper panels show dorso-lateral corazonin-producing neurons (dlCrz) and lower panels show ventral nerve cord corazonin-producing neurons (vCrz). for quantitative analyses of (B) and (D).

Techniques Used: Staining, Immunostaining

8) Product Images from "Glucuronylated core 1 glycans are required for precise localization of neuromuscular junctions and normal formation of basement membranes on Drosophila muscles"

Article Title: Glucuronylated core 1 glycans are required for precise localization of neuromuscular junctions and normal formation of basement membranes on Drosophila muscles

Journal: Developmental biology

doi: 10.1016/j.ydbio.2018.02.017

Genetic interaction between dC1GalT1 and dGlcAT-P (A–F″) Confocal images of basement membranes and NMJ boutons on muscle 6 (M6) and muscle 7 (M7) at abdominal segment 3 in dC1GalT1 2.1 /+ (A–B″), dGlcAT-P SK6 /+ (C–D″), and dC1GalT1 2.1 /+; dGlcAT-P SK6 /+ (E–F″) third-instar larvae. Surface sectional views of the muscles are presented in A–F. Internal sectional views of the muscles are presented in A′–F′. Cross-sectional views of the areas within white dotted-lines in A–F are presented in A″–F″. Basement membranes and NMJ boutons were labeled with an anti-nidogen (Ndg) antibody (magenta) and an anti-fasciclin II (Fas II) antibody (green), respectively. Muscle fibers and cell nuclei (DNA) were labeled with phalloidin (white) and Hoechst 33342 (blue), respectively. White arrowheads in E/F and E′/F′ indicate mislocalized NMJ boutons and a partial loss of Ndg, respectively. Black arrowheads in A″– F″ show positions of NMJ boutons. Scale bars: 50 µm (A–F′) and 10 µm (A″–F″). (G) Frequency of Ndg loss phenotype in dC1GalT1 2.1 /+ ( n = 19), dGlcAT-P SK6 /+ ( n = 20), and dC1GalT1 21 /+; dGlcAT-P SK6 /+ ( n = 21) larvae. (H) Total range of Ndg staining loss at the muscle 6/7 boundary in dC1GalT1 2.1 /+ ( n = 19), dGlcAT-P SK6 /+ ( n = 20), and dC1GalT1 2.1 /+; dGlcAT-P SK6 /+ ( n = 21) larvae. Statistical significance was assessed by Steel-Dwass test. * P
Figure Legend Snippet: Genetic interaction between dC1GalT1 and dGlcAT-P (A–F″) Confocal images of basement membranes and NMJ boutons on muscle 6 (M6) and muscle 7 (M7) at abdominal segment 3 in dC1GalT1 2.1 /+ (A–B″), dGlcAT-P SK6 /+ (C–D″), and dC1GalT1 2.1 /+; dGlcAT-P SK6 /+ (E–F″) third-instar larvae. Surface sectional views of the muscles are presented in A–F. Internal sectional views of the muscles are presented in A′–F′. Cross-sectional views of the areas within white dotted-lines in A–F are presented in A″–F″. Basement membranes and NMJ boutons were labeled with an anti-nidogen (Ndg) antibody (magenta) and an anti-fasciclin II (Fas II) antibody (green), respectively. Muscle fibers and cell nuclei (DNA) were labeled with phalloidin (white) and Hoechst 33342 (blue), respectively. White arrowheads in E/F and E′/F′ indicate mislocalized NMJ boutons and a partial loss of Ndg, respectively. Black arrowheads in A″– F″ show positions of NMJ boutons. Scale bars: 50 µm (A–F′) and 10 µm (A″–F″). (G) Frequency of Ndg loss phenotype in dC1GalT1 2.1 /+ ( n = 19), dGlcAT-P SK6 /+ ( n = 20), and dC1GalT1 21 /+; dGlcAT-P SK6 /+ ( n = 21) larvae. (H) Total range of Ndg staining loss at the muscle 6/7 boundary in dC1GalT1 2.1 /+ ( n = 19), dGlcAT-P SK6 /+ ( n = 20), and dC1GalT1 2.1 /+; dGlcAT-P SK6 /+ ( n = 21) larvae. Statistical significance was assessed by Steel-Dwass test. * P

Techniques Used: Labeling, Staining

Mislocalization of NMJ boutons at the muscle 6/7 boundary in dGlcAT-P null mutants (A–H″) Confocal images of basement membranes and NMJ boutons on muscle 6 (M6) and muscle 7 (M7) at abdominal segment 3 in wild-type (WT) (A–B″), dGlcAT-P null mutant (dGlcAT-P SK1 /Df(3L)BSC817 (C–D″), dGlcAT-P SK6 /Df(3L)BSC817 (E–F″)), and Act5C-Gal4 rescued ( Act5C-Gal4/UAS-dGlcAT-P; dGlcAT-P SK6 /Df(3L)BSC817) (G–H″) third-instar larvae. Surface sectional views of the muscles are presented in A–H. Internal sectional views of the muscles are presented in A′–H′. Cross-sectional views of the areas within white dotted-lines in A–H are presented in A″–H″. Basement membranes and NMJ boutons were labeled with an anti-nidogen (Ndg) antibody (magenta) and an anti-fasciclin II (Fas II) antibody (a presynaptic marker; green), respectively. Muscle fibers and cell nuclei (DNA) were labeled with phalloidin (white) and Hoechst 33342 (blue), respectively. Brackets in A, B, G, and H show the distance from the bouton to the muscle 6/7 boundary. White arrowheads in C–F and C′–F′ indicate mislocalized NMJ boutons and a partial loss of Ndg staining at the muscle 6/7 boundary, respectively. Black arrowheads in A″–H″ show positions of NMJ boutons. Scale bars: 50 µm (A–H′) and 10 µm (A″–H″). (I) Distance between each bouton and muscle 6/7 boundary in WT ( n = 32), dGlcAT-P SK1 /Df(3L)BSC817 ( n = 27), dGlcAT-P SK6 /Df(3L)BSC817 ( n = 28), and Act5C-Gal4 rescued ( n = 25) larvae. Statistical significance was assessed by the Steel-Dwass test. ** P
Figure Legend Snippet: Mislocalization of NMJ boutons at the muscle 6/7 boundary in dGlcAT-P null mutants (A–H″) Confocal images of basement membranes and NMJ boutons on muscle 6 (M6) and muscle 7 (M7) at abdominal segment 3 in wild-type (WT) (A–B″), dGlcAT-P null mutant (dGlcAT-P SK1 /Df(3L)BSC817 (C–D″), dGlcAT-P SK6 /Df(3L)BSC817 (E–F″)), and Act5C-Gal4 rescued ( Act5C-Gal4/UAS-dGlcAT-P; dGlcAT-P SK6 /Df(3L)BSC817) (G–H″) third-instar larvae. Surface sectional views of the muscles are presented in A–H. Internal sectional views of the muscles are presented in A′–H′. Cross-sectional views of the areas within white dotted-lines in A–H are presented in A″–H″. Basement membranes and NMJ boutons were labeled with an anti-nidogen (Ndg) antibody (magenta) and an anti-fasciclin II (Fas II) antibody (a presynaptic marker; green), respectively. Muscle fibers and cell nuclei (DNA) were labeled with phalloidin (white) and Hoechst 33342 (blue), respectively. Brackets in A, B, G, and H show the distance from the bouton to the muscle 6/7 boundary. White arrowheads in C–F and C′–F′ indicate mislocalized NMJ boutons and a partial loss of Ndg staining at the muscle 6/7 boundary, respectively. Black arrowheads in A″–H″ show positions of NMJ boutons. Scale bars: 50 µm (A–H′) and 10 µm (A″–H″). (I) Distance between each bouton and muscle 6/7 boundary in WT ( n = 32), dGlcAT-P SK1 /Df(3L)BSC817 ( n = 27), dGlcAT-P SK6 /Df(3L)BSC817 ( n = 28), and Act5C-Gal4 rescued ( n = 25) larvae. Statistical significance was assessed by the Steel-Dwass test. ** P

Techniques Used: Mutagenesis, Labeling, Marker, Staining

9) Product Images from "Coordinated movement, neuromuscular synaptogenesis and trans-synaptic signaling defects in Drosophila galactosemia models"

Article Title: Coordinated movement, neuromuscular synaptogenesis and trans-synaptic signaling defects in Drosophila galactosemia models

Journal: Human Molecular Genetics

doi: 10.1093/hmg/ddw217

dGALE and dUGP mutants manifest glycan losses in the NMJ synaptomatrix. Representative NMJs co-labelled with anti-Fasciclin II (FASII, red). ( A ) Wisteria floribunda lectin (WFA, green) or ( B ) anti-horseradish peroxidase (HRP; green) in genetic control
Figure Legend Snippet: dGALE and dUGP mutants manifest glycan losses in the NMJ synaptomatrix. Representative NMJs co-labelled with anti-Fasciclin II (FASII, red). ( A ) Wisteria floribunda lectin (WFA, green) or ( B ) anti-horseradish peroxidase (HRP; green) in genetic control

Techniques Used:

10) Product Images from "Mcm3 replicative helicase mutation impairs neuroblast proliferation and memory in Drosophila"

Article Title: Mcm3 replicative helicase mutation impairs neuroblast proliferation and memory in Drosophila

Journal: Genes, brain, and behavior

doi: 10.1111/gbb.12304

Reduced proliferation potential of single MBNBs in smu (a) MARCM analysis of the cell lineage derived from a single MBNB in a control animal. Clones were induced at 1 st ). This is indicated by the projection pattern of the mCD8∷GFP-labeled axons (green) in the different lobes of the adult MB. Fasciclin II staining (red) highlights the α/β-lobes and is weakly detected in γ- and α′/β′-lobes. (b) smu mutant MBNBs precociously stop proliferation before generating the latest born α/β-neurons. Only a reduced number of γ-neurons (left brain hemisphere) or sometimes additionally few α′/β′-neurons (right brain hemisphere) are generated ( n = 12 single smu MBNB clones). Scale bar: 20 μm. (c-e) Quantification of MBNB-derived GMCs in 3 rd instar larval brains. (c) These GMCs express Tailless (Tll, red) and are surrounded by KCs labeled with Dachshund (Dac, blue). Worniu-Gal4 driven expression of UAS-mCD8∷GFP (green) marks central brain neuroblasts and, by perdurance of the protein, also outlines progeny cells. In controls, a cluster of GMCs is associated with each of the 4 MBNBs (encircled). Scale bar: 10 μm. (d) In smu , the number of Tll-positive GMC clusters was reduced, indicating loss of MBNBs or their inability to proliferate. (e) Compared to controls, also the number of GMCs in each cluster was reduced in smu . *** P ≤ 0.001.
Figure Legend Snippet: Reduced proliferation potential of single MBNBs in smu (a) MARCM analysis of the cell lineage derived from a single MBNB in a control animal. Clones were induced at 1 st ). This is indicated by the projection pattern of the mCD8∷GFP-labeled axons (green) in the different lobes of the adult MB. Fasciclin II staining (red) highlights the α/β-lobes and is weakly detected in γ- and α′/β′-lobes. (b) smu mutant MBNBs precociously stop proliferation before generating the latest born α/β-neurons. Only a reduced number of γ-neurons (left brain hemisphere) or sometimes additionally few α′/β′-neurons (right brain hemisphere) are generated ( n = 12 single smu MBNB clones). Scale bar: 20 μm. (c-e) Quantification of MBNB-derived GMCs in 3 rd instar larval brains. (c) These GMCs express Tailless (Tll, red) and are surrounded by KCs labeled with Dachshund (Dac, blue). Worniu-Gal4 driven expression of UAS-mCD8∷GFP (green) marks central brain neuroblasts and, by perdurance of the protein, also outlines progeny cells. In controls, a cluster of GMCs is associated with each of the 4 MBNBs (encircled). Scale bar: 10 μm. (d) In smu , the number of Tll-positive GMC clusters was reduced, indicating loss of MBNBs or their inability to proliferate. (e) Compared to controls, also the number of GMCs in each cluster was reduced in smu . *** P ≤ 0.001.

Techniques Used: Derivative Assay, Clone Assay, Labeling, Staining, Mutagenesis, Generated, Expressing

11) Product Images from "Overelaborated synaptic architecture and reduced synaptomatrix glycosylation in a Drosophila classic galactosemia disease model"

Article Title: Overelaborated synaptic architecture and reduced synaptomatrix glycosylation in a Drosophila classic galactosemia disease model

Journal: Disease Models & Mechanisms

doi: 10.1242/dmm.017137

sgl overexpression prevents dGALT motor, NMJ and HRP glycosylation defects. (A) Representative NMJs imaged with anti-horseradish-peroxidase (HRP; green) and anti-Discs-large (DLG; red) for control ( dGALT C2 ), dGALT null with driver alone ( dGALT ΔAP2 ; UH1/+), sgl overexpression ( UAS-sgl/UH1-Gal4 ) and dGALT null with sgl overexpression ( dGALT; UAS-sgl/UH1-Gal4 ). (B) Normalized time required for the wandering L3 to rollover for all four genotypes. (C) Quantification of synaptic bouton number and (D) inter-bouton spacing distance. (E) Sample NMJs imaged with anti-Fasciclin-II (FASII; red) and anti-HRP (green). (F) Normalized quantification of HRP intensity. Sample size: ≥seven NMJs. Error bars show s.e.m. with significance indicated: * P
Figure Legend Snippet: sgl overexpression prevents dGALT motor, NMJ and HRP glycosylation defects. (A) Representative NMJs imaged with anti-horseradish-peroxidase (HRP; green) and anti-Discs-large (DLG; red) for control ( dGALT C2 ), dGALT null with driver alone ( dGALT ΔAP2 ; UH1/+), sgl overexpression ( UAS-sgl/UH1-Gal4 ) and dGALT null with sgl overexpression ( dGALT; UAS-sgl/UH1-Gal4 ). (B) Normalized time required for the wandering L3 to rollover for all four genotypes. (C) Quantification of synaptic bouton number and (D) inter-bouton spacing distance. (E) Sample NMJs imaged with anti-Fasciclin-II (FASII; red) and anti-HRP (green). (F) Normalized quantification of HRP intensity. Sample size: ≥seven NMJs. Error bars show s.e.m. with significance indicated: * P

Techniques Used: Over Expression

Loss of dGALT activity compromises the NMJ glycosylated synaptomatrix. Representative wandering L3 NMJs imaged with anti-Fasciclin-II (FASII; red) in all cases and co-labeled with lectins (A) Erythrina cristagalli (ECL; green), (B) Wisteria floribunda (WFA; green) and (C) anti-horseradish-peroxidase (HRP; green) in genetic controls (precise-excision dGALT C2 ) and dGALT nulls (imprecise-excision dGALT ΔAP2 ). Quantification shows normalized ECL (≥15 NMJs) (A′), WFA (≥19 NMJs) (B′) and anti-HRP (≥16 NMJs) (C′) intensities. Error bars show s.e.m. with significance indicated as ** P
Figure Legend Snippet: Loss of dGALT activity compromises the NMJ glycosylated synaptomatrix. Representative wandering L3 NMJs imaged with anti-Fasciclin-II (FASII; red) in all cases and co-labeled with lectins (A) Erythrina cristagalli (ECL; green), (B) Wisteria floribunda (WFA; green) and (C) anti-horseradish-peroxidase (HRP; green) in genetic controls (precise-excision dGALT C2 ) and dGALT nulls (imprecise-excision dGALT ΔAP2 ). Quantification shows normalized ECL (≥15 NMJs) (A′), WFA (≥19 NMJs) (B′) and anti-HRP (≥16 NMJs) (C′) intensities. Error bars show s.e.m. with significance indicated as ** P

Techniques Used: Activity Assay, Labeling

Co-removal of dGALK prevents the loss of galactosylation in the dGALT -null synaptomatrix. Representative NMJs imaged with anti-Fasciclin-II (FASII; red) and (A) Erythrina cristagalli lectin (ECL; green) or (B) Wisteria floribunda agglutinin (WFA; green) in control ( dGALT C2 ), dGALK -null ( dGALK ΔEXC9 ), dGALT -null ( dGALT ΔAP2 ) and double-null ( dGALT ΔAP2 ; dGALK ΔEXC9 ) larvae. Normalized quantification of ECL (A′) and WFA (B′) intensities. Sample size: ≥six NMJs. Error bars show s.e.m. with significance indicated: *** P
Figure Legend Snippet: Co-removal of dGALK prevents the loss of galactosylation in the dGALT -null synaptomatrix. Representative NMJs imaged with anti-Fasciclin-II (FASII; red) and (A) Erythrina cristagalli lectin (ECL; green) or (B) Wisteria floribunda agglutinin (WFA; green) in control ( dGALT C2 ), dGALK -null ( dGALK ΔEXC9 ), dGALT -null ( dGALT ΔAP2 ) and double-null ( dGALT ΔAP2 ; dGALK ΔEXC9 ) larvae. Normalized quantification of ECL (A′) and WFA (B′) intensities. Sample size: ≥six NMJs. Error bars show s.e.m. with significance indicated: *** P

Techniques Used:

12) Product Images from "A Targeted Glycan-Related Gene Screen Reveals Heparan Sulfate Proteoglycan Sulfation Regulates WNT and BMP Trans-Synaptic Signaling"

Article Title: A Targeted Glycan-Related Gene Screen Reveals Heparan Sulfate Proteoglycan Sulfation Regulates WNT and BMP Trans-Synaptic Signaling

Journal: PLoS Genetics

doi: 10.1371/journal.pgen.1003031

Synaptic HSPG co-receptor abundance is modified by 6-O-S sulfation. (A) Representative NMJ synaptic boutons imaged from control ( w 1118 ), sulf1 and hs6st nulls, probed with presynaptic neural marker anti-HRP (green) and Dally-like (Dlp; red). Right: Dlp distribution without the HRP signal is shown for clarity. (B) Quantification of mean fluorescent intensity levels of anti-Dlp labeling normalized to the HRP co-label at the muscle 6 NMJ, normalized to genetic control. (C) Boutons labeled with neural marker anti-Fasciclin II (FasII, green) and anti-Syndecan (Sdc, red). Right: Sdc distribution is shown alone for clarity. (D) Quantification of the mean fluorescent intensity levels of anti-Sdc labeling at the muscle 6 NMJ, normalized to genetic control. Sample sizes are at least 12 independent NMJs of at least 7 animals per indicated genotypes. Statistically significant differences calculated using student's t-test, * p
Figure Legend Snippet: Synaptic HSPG co-receptor abundance is modified by 6-O-S sulfation. (A) Representative NMJ synaptic boutons imaged from control ( w 1118 ), sulf1 and hs6st nulls, probed with presynaptic neural marker anti-HRP (green) and Dally-like (Dlp; red). Right: Dlp distribution without the HRP signal is shown for clarity. (B) Quantification of mean fluorescent intensity levels of anti-Dlp labeling normalized to the HRP co-label at the muscle 6 NMJ, normalized to genetic control. (C) Boutons labeled with neural marker anti-Fasciclin II (FasII, green) and anti-Syndecan (Sdc, red). Right: Sdc distribution is shown alone for clarity. (D) Quantification of the mean fluorescent intensity levels of anti-Sdc labeling at the muscle 6 NMJ, normalized to genetic control. Sample sizes are at least 12 independent NMJs of at least 7 animals per indicated genotypes. Statistically significant differences calculated using student's t-test, * p

Techniques Used: Modification, Marker, Labeling

Loss of sulf1 and hs6st causes differential effects on BMP signaling. (A) Representative NMJ synaptic boutons on muscle 6 in segment A3 from control ( w 1118 ), sulf1 and hs6st nulls, labeled with neural marker anti-Fasciclin II (FasII, red) and for phosphorylated Mothers against decapentaplegic (P-Mad; green) activated downstream of Gbb signaling. Arrows indicate representative P-Mad punctae in the indicated genotypes. (B) Representative ventral nerve cord (VNC) midlines from the same 3 genotypes, labeled with anti-FasII (red) and P-Mad (green). Labeled motor neuron nuclei are indicated by arrows. Quantification of the mean fluorescent intensity level of P-Mad labeling normalized to FasII co-label at the NMJ synapse (C) and in motor neuron nuclei (D), normalized to genetic control. Sample sizes are ≥14 animals per indicated genotypes. Statistically significant differences calculated using the Mann-Whitney test for non-parametric data, * p
Figure Legend Snippet: Loss of sulf1 and hs6st causes differential effects on BMP signaling. (A) Representative NMJ synaptic boutons on muscle 6 in segment A3 from control ( w 1118 ), sulf1 and hs6st nulls, labeled with neural marker anti-Fasciclin II (FasII, red) and for phosphorylated Mothers against decapentaplegic (P-Mad; green) activated downstream of Gbb signaling. Arrows indicate representative P-Mad punctae in the indicated genotypes. (B) Representative ventral nerve cord (VNC) midlines from the same 3 genotypes, labeled with anti-FasII (red) and P-Mad (green). Labeled motor neuron nuclei are indicated by arrows. Quantification of the mean fluorescent intensity level of P-Mad labeling normalized to FasII co-label at the NMJ synapse (C) and in motor neuron nuclei (D), normalized to genetic control. Sample sizes are ≥14 animals per indicated genotypes. Statistically significant differences calculated using the Mann-Whitney test for non-parametric data, * p

Techniques Used: Labeling, Marker, MANN-WHITNEY

Synaptic WNT and BMP ligand abundance is modified by 6-O-S sulfation. Images show muscle 6 NMJ in segment A3 probed in non-detergent conditions, so that only extracellular protein distributions are detected. The white lines indicate cross-section planes for spatial measurements. Insets indicate single synaptic boutons at higher magnification. (A) Representative NMJ boutons from control ( w 1118 ), sulf1 and hs6st null genotypes, labeled for presynaptic anti-horseradish peroxidase (HRP, red) and anti-wingless (Wg, green). (B) Extracellular distribution of Wg across the diameter of a synaptic bouton. The Y-axis indicates intensity and the X-axis shows distance in microns. The HRP intensity profile is indicated in red; Wg intensity is shown in green. (C) Quantification of Wg mean intensity levels normalized to the HRP co-label, and to genetic control. Sample sizes are at least 15 animals per indicated genotypes. (D) Representative synaptic boutons labeled with presynaptic anti-Fasciclin II (FasII; green) and anti-Glass Bottom Boat (Gbb; red). (E) Gbb distribution across the diameter of a synaptic bouton. Y-axis indicates intensity and the X-axis shows distance in microns. FasII intensity profile is indicated in green; Gbb intensity is shown in red. (F) Quantification of Gbb mean intensity levels normalized to genetic control. Sample sizes are at least 11 independent NMJs of at least 7 animals per indicated genotypes. Statistically significant differences calculated using student's t-test and Mann-Whitney test for non-parametric data, ** p
Figure Legend Snippet: Synaptic WNT and BMP ligand abundance is modified by 6-O-S sulfation. Images show muscle 6 NMJ in segment A3 probed in non-detergent conditions, so that only extracellular protein distributions are detected. The white lines indicate cross-section planes for spatial measurements. Insets indicate single synaptic boutons at higher magnification. (A) Representative NMJ boutons from control ( w 1118 ), sulf1 and hs6st null genotypes, labeled for presynaptic anti-horseradish peroxidase (HRP, red) and anti-wingless (Wg, green). (B) Extracellular distribution of Wg across the diameter of a synaptic bouton. The Y-axis indicates intensity and the X-axis shows distance in microns. The HRP intensity profile is indicated in red; Wg intensity is shown in green. (C) Quantification of Wg mean intensity levels normalized to the HRP co-label, and to genetic control. Sample sizes are at least 15 animals per indicated genotypes. (D) Representative synaptic boutons labeled with presynaptic anti-Fasciclin II (FasII; green) and anti-Glass Bottom Boat (Gbb; red). (E) Gbb distribution across the diameter of a synaptic bouton. Y-axis indicates intensity and the X-axis shows distance in microns. FasII intensity profile is indicated in green; Gbb intensity is shown in red. (F) Quantification of Gbb mean intensity levels normalized to genetic control. Sample sizes are at least 11 independent NMJs of at least 7 animals per indicated genotypes. Statistically significant differences calculated using student's t-test and Mann-Whitney test for non-parametric data, ** p

Techniques Used: Modification, Labeling, MANN-WHITNEY

13) Product Images from "Drosophila rugose Is a Functional Homolog of Mammalian Neurobeachin and Affects Synaptic Architecture, Brain Morphology, and Associative Learning"

Article Title: Drosophila rugose Is a Functional Homolog of Mammalian Neurobeachin and Affects Synaptic Architecture, Brain Morphology, and Associative Learning

Journal: The Journal of Neuroscience

doi: 10.1523/JNEUROSCI.6424-11.2012

The morphological MB defects in rg mutants are genetically separable from the learning phenotype. Whole-mount anti-fasciclin II staining of the adult central brain labels α, β, γ MB lobes and the ellipsoid body (EB). A , Control w 1118 MB lobes (α, β, γ) and EB. B , rg 1 MBs show only minor defects compared with control brains. C , rg γ 5 hemizygote with an aberrant morphology of the β lobe and β-lobe fusion. D , Overextension of the β lobe and full β-lobe misguidance in rg FDD brains. E , Full β misguidance and aberrant γ-lobe morphology of rg FDD MBs. MB defects mentioned for C–E are indicated with arrows. F–H , Quantification of the frequency of the morphological defects in rg mutants ( F ) and their respective rescues in rg γ 5 ( G ) and rg FDD ( H ) background combined with a Gal4 driver. rg γ 5 and rg FDD combined with the Gal4 driver were used as mutant controls (indicated in gray) to correct for effects of the Gal4 driver and genetic background. The rescues with the rugose transgene generally rescued the phenotypes completely or partially with, respectively, elav–Gal4 or the MB driver OK107–Gal4. Reintroducing rg + cDNA with MB247–Gal4 had no effect. Nbea cDNA expression failed to rescue the phenotypes. Numbers represent the percentages of aberrant hemispheres. Fusion defects were counted as defects of two hemispheres. n is the number of brain hemispheres analyzed. Statistical significance between rg γ 5 or rg FDD combined with a Gal4 driver and the respective rescues with transgenic rg + or Nbea cDNA were assessed with the Fisher's exact test. For rugose mutants, statistical significance with w 1118 brains was also assessed with the Fisher's exact test.
Figure Legend Snippet: The morphological MB defects in rg mutants are genetically separable from the learning phenotype. Whole-mount anti-fasciclin II staining of the adult central brain labels α, β, γ MB lobes and the ellipsoid body (EB). A , Control w 1118 MB lobes (α, β, γ) and EB. B , rg 1 MBs show only minor defects compared with control brains. C , rg γ 5 hemizygote with an aberrant morphology of the β lobe and β-lobe fusion. D , Overextension of the β lobe and full β-lobe misguidance in rg FDD brains. E , Full β misguidance and aberrant γ-lobe morphology of rg FDD MBs. MB defects mentioned for C–E are indicated with arrows. F–H , Quantification of the frequency of the morphological defects in rg mutants ( F ) and their respective rescues in rg γ 5 ( G ) and rg FDD ( H ) background combined with a Gal4 driver. rg γ 5 and rg FDD combined with the Gal4 driver were used as mutant controls (indicated in gray) to correct for effects of the Gal4 driver and genetic background. The rescues with the rugose transgene generally rescued the phenotypes completely or partially with, respectively, elav–Gal4 or the MB driver OK107–Gal4. Reintroducing rg + cDNA with MB247–Gal4 had no effect. Nbea cDNA expression failed to rescue the phenotypes. Numbers represent the percentages of aberrant hemispheres. Fusion defects were counted as defects of two hemispheres. n is the number of brain hemispheres analyzed. Statistical significance between rg γ 5 or rg FDD combined with a Gal4 driver and the respective rescues with transgenic rg + or Nbea cDNA were assessed with the Fisher's exact test. For rugose mutants, statistical significance with w 1118 brains was also assessed with the Fisher's exact test.

Techniques Used: Staining, Mutagenesis, Expressing, Transgenic Assay

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    Developmental Studies Hybridoma Bank mouse anti fasciclin ii
    Neuron-specific depletion of Edis causes abnormal axonogenesis and neurodevelopment. (A) Confocal images of dorsal anterior regions of adult brains with neuron-specific expression of control shgfp (left panel) or shEdis (middle and right panels) driven by Elav-Gal4 driver. In control brain (left panel), <t>anti-FasII</t> antibody delineates the central complex (CC) as well as the vertical α and horizontal β and γ lobes of the mushroom bodies ( MB s), with γ lobes showing weaker FasII signal, as indicated. Depletion of Edis resulted in a spectrum of severe, age-dependent morphological defects in the MBs. In middle panel, one α lobe and one β lobe were mostly missing, and the remaining α and β lobes showed abnormal morphology. In right panel, both α lobes were largely missing, while the two β lobes crossed the midline and merged together. Scale bar: 20 μm. (B) Quantification of MB morphology phenotypes shown in A (Chi-squared test, sample numbers are shown on top, *** p
    Mouse Anti Fasciclin Ii, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Developmental Studies Hybridoma Bank mouse anti fasciclin ii antibody
    Neuron-specific depletion of Edis causes abnormal axonogenesis and neurodevelopment. (A) Confocal images of dorsal anterior regions of adult brains with neuron-specific expression of control shgfp (left panel) or shEdis (middle and right panels) driven by Elav-Gal4 driver. In control brain (left panel), <t>anti-FasII</t> antibody delineates the central complex (CC) as well as the vertical α and horizontal β and γ lobes of the mushroom bodies ( MB s), with γ lobes showing weaker FasII signal, as indicated. Depletion of Edis resulted in a spectrum of severe, age-dependent morphological defects in the MBs. In middle panel, one α lobe and one β lobe were mostly missing, and the remaining α and β lobes showed abnormal morphology. In right panel, both α lobes were largely missing, while the two β lobes crossed the midline and merged together. Scale bar: 20 μm. (B) Quantification of MB morphology phenotypes shown in A (Chi-squared test, sample numbers are shown on top, *** p
    Mouse Anti Fasciclin Ii Antibody, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti fasciclin ii antibody/product/Developmental Studies Hybridoma Bank
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    Developmental Studies Hybridoma Bank mouse anti fasii
    Phenotypic analysis of the Ptr null mutant in whole mount embryos. Z-stack immunofluorescences against PNA and <t>FASII</t> (the two upper rows), PNA and Repo (the two middle rows) or PNA and 22C10 (the two bottom rows). The analysis was carried out on D. melanogaster late embryos (stage 15/16). Arrows indicate the alterations in the primary tracts of Ptr null mutants that were evidenced with PNA. The right column shows a higher magnification of the alterations found inside the dashed boxes in each image. In all the figures, embryos are oriented anterior to the left. To observe the nervous system, the first four rows of images are ventral views of the embryo, whereas, in the last two rows, the ventral side of the embryo is down. Bars: 50 μm.
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    Neuron-specific depletion of Edis causes abnormal axonogenesis and neurodevelopment. (A) Confocal images of dorsal anterior regions of adult brains with neuron-specific expression of control shgfp (left panel) or shEdis (middle and right panels) driven by Elav-Gal4 driver. In control brain (left panel), anti-FasII antibody delineates the central complex (CC) as well as the vertical α and horizontal β and γ lobes of the mushroom bodies ( MB s), with γ lobes showing weaker FasII signal, as indicated. Depletion of Edis resulted in a spectrum of severe, age-dependent morphological defects in the MBs. In middle panel, one α lobe and one β lobe were mostly missing, and the remaining α and β lobes showed abnormal morphology. In right panel, both α lobes were largely missing, while the two β lobes crossed the midline and merged together. Scale bar: 20 μm. (B) Quantification of MB morphology phenotypes shown in A (Chi-squared test, sample numbers are shown on top, *** p

    Journal: PLOS Genetics

    Article Title: The circular RNA Edis regulates neurodevelopment and innate immunityA circular RNA Edis-Relish-castor axis regulates neuronal development in Drosophila

    doi: 10.1371/journal.pgen.1010429

    Figure Lengend Snippet: Neuron-specific depletion of Edis causes abnormal axonogenesis and neurodevelopment. (A) Confocal images of dorsal anterior regions of adult brains with neuron-specific expression of control shgfp (left panel) or shEdis (middle and right panels) driven by Elav-Gal4 driver. In control brain (left panel), anti-FasII antibody delineates the central complex (CC) as well as the vertical α and horizontal β and γ lobes of the mushroom bodies ( MB s), with γ lobes showing weaker FasII signal, as indicated. Depletion of Edis resulted in a spectrum of severe, age-dependent morphological defects in the MBs. In middle panel, one α lobe and one β lobe were mostly missing, and the remaining α and β lobes showed abnormal morphology. In right panel, both α lobes were largely missing, while the two β lobes crossed the midline and merged together. Scale bar: 20 μm. (B) Quantification of MB morphology phenotypes shown in A (Chi-squared test, sample numbers are shown on top, *** p

    Article Snippet: Primary antibodies: mouse anti-Fasciclin II (1D4, Developmental Studies Hybridoma Bank, developed by Dr. Corey Goodman) at 1:50 dilution, rat anti-Elav antibody (7E8A10, Developmental Studies Hybridoma Bank, developed by Dr. Gerald M. Rubin) at 1:100 dilution, prepared in 1XPBT with 5% normal donkey serum to block nonspecific binding.

    Techniques: Expressing

    Lateral longitudinal axons labelled by GMR28F02 and GMR28G05 are distinct from FasII-positive lateral axon pathways.

    Journal: Fly

    Article Title: Characterization of enhancer fragments in Drosophila robo2

    doi: 10.1080/19336934.2022.2126259

    Figure Lengend Snippet: Lateral longitudinal axons labelled by GMR28F02 and GMR28G05 are distinct from FasII-positive lateral axon pathways.

    Article Snippet: The following antibodies were used: rabbit anti-GFP (Invitrogen #A11122, 1:1000), rabbit anti-c-Myc (Sigma #C3956), mouse anti-Fasciclin II (Developmental Studies Hybridoma Bank [DSHB] #1D4, 1:100), mouse anti-HA (BioLegend #901503, 1:1000), mouse anti-wrapper (DSHB #10D3, 1:100), HRP-conjugated goat anti-mouse (Jackson ImmunoResearch #115-035-003, 1:500), HRP-conjugated goat anti-rabbit (Jackson #111-035-003, 1:500), Alexa 488-conjugated goat anti-HRP (Jackson #123-545-021, 1:200), Alexa 647-conjugated goat anti-HRP (Jackson #123-605-021, 1:100), Cy3-conjugated goat anti-mouse (Jackson #115-165-003, 1:1000), Cy3-conjugated goat anti-rabbit (Jackson #111-165-003, 1:500), Alexa 647-conjugated goat anti-mouse (Jackson #115-605-003, 1:500), Alexa 488-conjugated goat anti-rabbit (Jackson #111-545-003, 1:500), Alexa 647-conjugated goat anti-rabbit (Jackson #111-605-144, 1:500).

    Techniques:

    Phenotypic analysis of the Ptr null mutant in whole mount embryos. Z-stack immunofluorescences against PNA and FASII (the two upper rows), PNA and Repo (the two middle rows) or PNA and 22C10 (the two bottom rows). The analysis was carried out on D. melanogaster late embryos (stage 15/16). Arrows indicate the alterations in the primary tracts of Ptr null mutants that were evidenced with PNA. The right column shows a higher magnification of the alterations found inside the dashed boxes in each image. In all the figures, embryos are oriented anterior to the left. To observe the nervous system, the first four rows of images are ventral views of the embryo, whereas, in the last two rows, the ventral side of the embryo is down. Bars: 50 μm.

    Journal: Frontiers in Neuroscience

    Article Title: Patched-Related Is Required for Proper Development of Embryonic Drosophila Nervous System

    doi: 10.3389/fnins.2022.920670

    Figure Lengend Snippet: Phenotypic analysis of the Ptr null mutant in whole mount embryos. Z-stack immunofluorescences against PNA and FASII (the two upper rows), PNA and Repo (the two middle rows) or PNA and 22C10 (the two bottom rows). The analysis was carried out on D. melanogaster late embryos (stage 15/16). Arrows indicate the alterations in the primary tracts of Ptr null mutants that were evidenced with PNA. The right column shows a higher magnification of the alterations found inside the dashed boxes in each image. In all the figures, embryos are oriented anterior to the left. To observe the nervous system, the first four rows of images are ventral views of the embryo, whereas, in the last two rows, the ventral side of the embryo is down. Bars: 50 μm.

    Article Snippet: Antibodies employed were: mouse anti-22C10 [1:50 22C10, Developmental Studies Hybridoma Bank (DSHB, IA, USA)] or mouse anti-FasII (1D4 anti-Fasciclin II, DSHB, diluted 1:30) or mouse anti-Repo (1:20 8D12 anti-Repo, DSHB).

    Techniques: Mutagenesis