Journal: Life Science Alliance

Article Title: Neuronal Aβ42 is enriched in small vesicles at the presynaptic side of synapses

doi: 10.26508/lsa.201800028

Figure Lengend Snippet: (A) Hippocampal neurons stained with Aβ42 (red) and synaptophysin (green), using the same primary and secondary antibodies as for STED images in combined with actin (light grey) staining and imaged by confocal microscopy. Scale bars: 5 μm. (B) Negative control for synaptophysin (syp neg cntr). Hippocampal neurons only stained with Aβ42 (red) and the same secondary antibodies (Abberior STAR 635P and Alexa Fluor 594) as for STED images in combined with actin (light grey) staining and imaged by confocal microscopy in the same setting as in (A). Scale bars: 5 μm. (C) Hippocampal neurons stained for Aβ42 (red) and PSD95 (cyan), using the same primary and secondary antibodies as for STED in combined with actin (light grey) staining and imaged by confocal microscopy. Scale bars: 5 μm. (D) Negative control for PSD95 (PSD95 neg cntr). Hippocampal neurons only stained for Aβ42 (red), using the same secondary antibodies (Abberrior STAR 635P and Alexa Fluor 594) as for STED images in combined with actin (light grey) staining and imaged by confocal microscopy in the same setting as in (C). Scale bars: 5 μm. (E) Negative control for Aβ42 (Aβ42 neg cntr). Hippocampal neurons only stained for PSD95 (cyan), using the same secondary antibodies (Abberrior STAR 635P and Alexa Fluor 594) as for STED in combined with actin (light grey) staining and imaged by confocal microscopy in the same setting as in (C). Scale bars: 5 μm.

Article Snippet: Mouse anti-PSD95 IgG, here denoted anti-PSD95 (Ab2723; Abcam), was used as a postsynaptic marker whereas mouse anti-synaptophysin IgG, here denoted anti-synaptophysin (SP15; Enzo Life Sciences), was used as a presynaptic marker.

Techniques: Staining, Confocal Microscopy, Negative Control

Journal: Life Science Alliance

Article Title: Neuronal Aβ42 is enriched in small vesicles at the presynaptic side of synapses

doi: 10.26508/lsa.201800028

Figure Lengend Snippet: (A, B) Hippocampal neurons stained to visualize Aβ42 (red), synaptophysin (green), and actin (light grey) were imaged by confocal microscopy (A) and STED microscopy (B). Yellow arrows point at enrichment of Aβ42 at the synapse. Left scale bars: 10 μm. Other scale bars: 1 μm. (C, D) Hippocampal neurons subjected to immunocytochemistry to visualize Aβ42 (red), PSD95 (cyan), and actin (light grey) were imaged by confocal microscopy (C) and STED microscopy (D). Yellow arrows point at enrichment of Aβ42 at the synapse. Green arrow points at an Aβ42-stained vesicle where Aβ42 is stained only at the rim of the vesicle. Left scale bars: 4 μm. Other scale bars: 1 μm.

Article Snippet: Mouse anti-PSD95 IgG, here denoted anti-PSD95 (Ab2723; Abcam), was used as a postsynaptic marker whereas mouse anti-synaptophysin IgG, here denoted anti-synaptophysin (SP15; Enzo Life Sciences), was used as a presynaptic marker.

Techniques: Staining, Confocal Microscopy, Microscopy, Immunocytochemistry

Journal: Life Science Alliance

Article Title: Neuronal Aβ42 is enriched in small vesicles at the presynaptic side of synapses

doi: 10.26508/lsa.201800028

Figure Lengend Snippet: (A) STED image showing Aβ42 (red) and synaptophysin (green) combined with a confocal channel for actin (light grey). The left image has a lower brightness in the actin channel than the right image. Note that the axonal staining appears for many axons only in the right image with increased brightness. This image was further zoomed-in in to focus on the synapses. Scale bars: 2 μm. (B) STED image showing Aβ42 (red) and PSD95 (cyan) combined with a confocal channel for actin (light grey). The left image has a lower brightness in the actin channel than in the right channel. Note the intensive axon network both between and along the neurites. Scale bars: 2 μm.

Article Snippet: Mouse anti-PSD95 IgG, here denoted anti-PSD95 (Ab2723; Abcam), was used as a postsynaptic marker whereas mouse anti-synaptophysin IgG, here denoted anti-synaptophysin (SP15; Enzo Life Sciences), was used as a presynaptic marker.

Techniques: Staining

Journal: Life Science Alliance

Article Title: Neuronal Aβ42 is enriched in small vesicles at the presynaptic side of synapses

doi: 10.26508/lsa.201800028

Figure Lengend Snippet: (A, B) Zoomed STED images of synapses showing Aβ42 (red) and the presynaptic marker synaptophysin (green). Neuronal structure is shown by actin in a confocal channel (light grey) merged to the STED channels in the left panel. The staining of synaptophysin and Aβ42 at presynaptic boutons connected to the postsynaptic spines are shown by the yellow arrows. The staining of synaptophysin and Aβ42 at “free” boutons that do not connect to the postsynaptic spine is shown by a pink arrow. Scale bars: 1 μm. (C, D) Zoomed STED images of synapses showing Aβ42 (red) and the postsynaptic marker PSD95 (cyan). Neuronal structure is shown by actin staining in a confocal channel (light grey) merged to the STED channels in the left panel. Yellow arrows point at Aβ42 vesicles in axonal regions opposite to PSD95-containing stubby spines in (A) and to Aβ42 vesicles in axonal regions opposite to a mushroom spine in (B). Scale bars: 1 μm. (E) Quantification of Aβ42 staining in pre- and postsynapses. The staining of Aβ42 combined with the presynaptic marker synaptophysin (green bar) or postsynaptic marker PSD95 (cyan bar) was quantified from STED data. Graphpad prism was used to prepare graphs showing the proportion ± SD of presynapses (n = 73) and postsynapses (n = 73) that were positively stained by Aβ42.

Article Snippet: Mouse anti-PSD95 IgG, here denoted anti-PSD95 (Ab2723; Abcam), was used as a postsynaptic marker whereas mouse anti-synaptophysin IgG, here denoted anti-synaptophysin (SP15; Enzo Life Sciences), was used as a presynaptic marker.

Techniques: Marker, Staining

Journal: Life Science Alliance

Article Title: Neuronal Aβ42 is enriched in small vesicles at the presynaptic side of synapses

doi: 10.26508/lsa.201800028

Figure Lengend Snippet: (A, B) Zoomed STED images of synapses showing Aβ42 (red) and the presynaptic marker synaptophysin (green) combined with actin in a confocal channel (light grey) in the left panel. The staining of synaptophysin and Aβ42 at presynaptic boutons connected to the postsynaptic spines are shown by the yellow arrows. Scale bars: 1 μm. (C, D) Zoomed STED images of synapses showing Aβ42 (red) and the presynaptic marker PSD95 (cyan) combined with actin in a confocal channel (light grey) in the left panel. The staining of PSD95 at postsynaptic spines and Aβ42 at presynaptic boutons is shown (yellow arrows). Scale bars: 1 μm.

Article Snippet: Mouse anti-PSD95 IgG, here denoted anti-PSD95 (Ab2723; Abcam), was used as a postsynaptic marker whereas mouse anti-synaptophysin IgG, here denoted anti-synaptophysin (SP15; Enzo Life Sciences), was used as a presynaptic marker.

Techniques: Marker, Staining

Journal: Life Science Alliance

Article Title: Neuronal Aβ42 is enriched in small vesicles at the presynaptic side of synapses

doi: 10.26508/lsa.201800028

Figure Lengend Snippet: Colocalization between Aβ42 or synaptobrevin with synaptophysin staining in presynapses. The percentage of presynaptic marker synaptophysin (green) staining combined with Aβ42 staining (red) or another presynaptic marker synaptobrevin (red) was quantified from STED data using imageJ based on the area of staining using Otsu thresholding. Meta-chart was used to prepare Venn charts. 50 regions of interest were quantified for each staining combination. Total stained pixels in each region of interest was set to 100%.

Article Snippet: Mouse anti-PSD95 IgG, here denoted anti-PSD95 (Ab2723; Abcam), was used as a postsynaptic marker whereas mouse anti-synaptophysin IgG, here denoted anti-synaptophysin (SP15; Enzo Life Sciences), was used as a presynaptic marker.

Techniques: Staining, Marker

Journal: Life Science Alliance

Article Title: Neuronal Aβ42 is enriched in small vesicles at the presynaptic side of synapses

doi: 10.26508/lsa.201800028

Figure Lengend Snippet: (A) dSTORM images show Aβ42 (red) and synaptophysin (green). Two different presynaptic regions from the same image are shown in the left and right image. Note that the Aβ42 staining appears as vesicles that are interspersed in the same region as synaptophysin. Scale bars: 500 nm. (B) STORM images show Aβ42 (red) and PSD95 (cyan). Two different postsynaptic regions from the same image are shown in the left and right image. Note the proximity between Aβ42 and PSD95, in agreement with a synaptic cleft between the two stains. The somewhat overlap in the left panel is probably related to the fact that the synapse is not in the plane of the image. Scale bars: 500 nm.

Article Snippet: Mouse anti-PSD95 IgG, here denoted anti-PSD95 (Ab2723; Abcam), was used as a postsynaptic marker whereas mouse anti-synaptophysin IgG, here denoted anti-synaptophysin (SP15; Enzo Life Sciences), was used as a presynaptic marker.

Techniques: Staining

Journal: Life Science Alliance

Article Title: Neuronal Aβ42 is enriched in small vesicles at the presynaptic side of synapses

doi: 10.26508/lsa.201800028

Figure Lengend Snippet: STED images stained with Aβ42 (red) and synaptophysin (syp, green) combined with an actin (light grey) confocal channel. The yellow circles show synapses selected for quantification: 73 in total. Scale bars: 5 μm.

Article Snippet: Mouse anti-PSD95 IgG, here denoted anti-PSD95 (Ab2723; Abcam), was used as a postsynaptic marker whereas mouse anti-synaptophysin IgG, here denoted anti-synaptophysin (SP15; Enzo Life Sciences), was used as a presynaptic marker.

Techniques: Staining

Journal: Oncotarget

Article Title: Long-term neurocognitive dysfunction in offspring via NGF/ ERK/CREB signaling pathway caused by ketamine exposure during the second trimester of pregnancy in rats

doi: 10.18632/oncotarget.16042

Figure Lengend Snippet: Ketamine administered during the second trimester perturbed NGF and down-regulated ERK, p-ERK, MSK, CREB, p-CREB, SYP, SYN, and PSD95 in the hippocampi of offspring. On postnatal day 30, after behavioral tests, brains were harvested for western blot analyses to determine the expression of NGF, ERK, p-ERK, MSK, CREB, p-CREB, SYP, SYN, and PSD95. (A–I ) Above are representative immunoblots for the expression levels of the corresponding proteins, and below are the histograms of the quantification of the corresponding proteins normalized to GAPDH. Data are expressed as mean percentage ± SEM of control mean values. * p < 0.05.

Article Snippet: The separated proteins were transferred onto nitrocellulose (NC) membranes and incubated with rabbit anti- GAPDH, NGF, ERK, p-ERK, CREB, p-CREB, MSK1, SYP, SYN, and PSD95 (1:600, EnoGene, Nanjing, China) overnight at a temperature of 4 °C.

Techniques: Western Blot, Expressing, Control

Journal: Oncotarget

Article Title: Long-term neurocognitive dysfunction in offspring via NGF/ ERK/CREB signaling pathway caused by ketamine exposure during the second trimester of pregnancy in rats

doi: 10.18632/oncotarget.16042

Figure Lengend Snippet: Ketamine administered during the second trimester down-regulated ERK, p-ERK, MSK, CREB, p-CREB, SYP, SYN, and PSD95 in PC12 cells during in vitro experiments. SCH772984 is the inhibitor of the ERK protein. Through the contrast between S group and C group, a very good effect of the inhibitor is revealed. (A) Representative immunoblots for expression levels of the corresponding proteins. (B–I ) Histograms that quantify the corresponding proteins normalized to GAPDH, in which the data are expressed as mean percentages ± SEM of control mean values. * p < 0.05

Article Snippet: The separated proteins were transferred onto nitrocellulose (NC) membranes and incubated with rabbit anti- GAPDH, NGF, ERK, p-ERK, CREB, p-CREB, MSK1, SYP, SYN, and PSD95 (1:600, EnoGene, Nanjing, China) overnight at a temperature of 4 °C.

Techniques: In Vitro, Western Blot, Expressing, Control

Journal: Neural Development

Article Title: Imp is expressed in INPs and newborn neurons where it regulates neuropil targeting in the central complex

doi: 10.1186/s13064-023-00177-9

Figure Lengend Snippet: Key Resource Table

Article Snippet: Antibody , Rabbit anti-Syp , Genescript (Piscataway, NJ) , #4060 , 1:1000.

Techniques: Knockdown, Over Expression, Membrane, Control, Labeling