Journal: STAR Protocols

Article Title: Volumetric super-resolution imaging by serial ultrasectioning and stochastic optical reconstruction microscopy in mouse neural tissue

doi: 10.1016/j.xpro.2021.100971

Figure Lengend Snippet:

Article Snippet: Guinea pig polyclonal anti-Calbindin (1:100) , Synaptic Systems , Cat#214 005.

Techniques: Recombinant, Saline, Electron Microscopy, Plasmid Preparation, Software, Imaging, Control, Mass Measurement, Capsules, Microscopy, Staining, Adhesive, Spectrophotometry

Journal: The Journal of Neuroscience

Article Title: Laminin β2 Chain Regulates Retinal Progenitor Cell Mitotic Spindle Orientation via Dystroglycan

doi: 10.1523/JNEUROSCI.0551-18.2018

Figure Lengend Snippet: Deletion of laminin β2 leads to overproduction of rod photoreceptors and underproduction of bipolar cells and Müller glia. A, B, Extended focus view of z stacks from OPL and adjacent part of INL in flat-mount retinas stained for calbindin (horizontal cells) and cone arrestin (cone photoreceptors). The numbers of horizontal cells and cone photoreceptors were determined by counting in retinal flat-mounts and were found to be unaffected in the Lamb2−/− retinas. C, D, Quantification of cone photoreceptor and horizontal cell densities. E, F, Syntaxin-1 staining of retinal sections labeling amacrine cells. G, Quantification of the amacrine cell numbers. H, I, Hoechst staining of retinal cross sections. J, Quantification of rows of cells in the photoreceptor layer. K, L, Chx10 (bipolar cells) and Sox9 (Müller glia) staining of retinal cross sections. M, N, Quantification of bipolar cells and Müller glia. Data are mean ± SEM. *p ≤ 0.05, **p ≤ 0.01.

Article Snippet: The following antibodies were used: mouse anti-Centrin (1:1000, Millipore, 04-1624), rat anti-phospho histone H3 (1:3000, Sigma-Aldrich, H9908), sheep anti-Chx10 (1:300, Abcam, ab16141), rabbit anti-Sox9 (1:300, Millipore, AB5535), mouse anti-phospho-vimentin (1:300, MBL, D076-3), rabbit anti-Olig2 (1:300, Millipore, AB9610), mouse anti-syntaxin (1:500, Sigma-Aldrich, S0664), rabbit anti-cone arrestin, mCAR-LUMIj (1:10,000), previously described ( Nikonov et al., 2008 ) and a kind gift from gift from Cheryl Craft (University of California–Los Angeles), mouse anti-α-DG (1:50, Millipore, 05-298), rabbit anti-β-DG (1:300, JAF), a kind gift from Dominique Mornet (Université de Montpellier), rat anti-β1-integrin (1:300, Millipore, MAB1997), and guinea pig anti-calbindin (1:600, Synaptic Systems, 214 005).

Techniques: Staining, Labeling

Journal: PLOS One

Article Title: Impaired dynein function preserves spinal interneuron survival and positioning in an ALS-like mouse model

doi: 10.1371/journal.pone.0346246

Figure Lengend Snippet: Number of ChAT + (A-C), GAD-67 + (D), Calbindin + (E), Parvalbumin + (F), Parvalbumin + −Calbindin + (G), Parvalbumin + −GAD-67 + (H), and ChAT + −GAD-67 + (I) cells per biological replicate. The number of cells analysed is the sum of immunostained positive cells from eight spinal cord cross-sections (two sections per lumbar level L3–L6) per biological replicate. Data are shown as mean ± standard deviation and were analysed by an ordinary one-way ANOVA. N = 3 biological replicates per genotype.

Article Snippet: Following three 10-min PBS washes at 100 rpm, secondary probing was performed for 3 hours in the dark in PBS containing 0.05% Tween-20 and the relevant secondary antibodies for ChAT (1:250 dilution, donkey anti-goat IgG H + L Alexa Fluor ® 405, Abcam, ab175664), Calbindin (1:500 dilution, Rhodamine TRITC-AffiniPure donkey anti-guinea pig Ig G H + L, Jackson Immuno Research, 706-025-148-JIR), Parvalbumin (1:500 dilution, Alexa Fluor 647-AffiniPure donkey anti-rabbit IgG H + L, Jackson Immuno Research, 711-605-152), and GAD-67 (1:500 dilution, donkey anti-chicken IgY H + L Highly Cross Adsorbed Alexa Fluor ® 488, Life Technologies, A78948).

Techniques: Standard Deviation

Journal: Frontiers in Molecular Neuroscience

Article Title: Connexin30.2: In Vitro Interaction with Connexin36 in HeLa Cells and Expression in AII Amacrine Cells and Intrinsically Photosensitive Ganglion Cells in the Mouse Retina

doi: 10.3389/fnmol.2016.00036

Figure Lengend Snippet: List of primary antibodies used in this study .

Article Snippet: Calretinin , Guinea pig calretinin, full length amino acid sequence , Goat, polyclonal , 1:500 , Merck-Millipore, Darmstadt, Germany (AB 1550).

Techniques: Recombinant, Sequencing, Derivative Assay, Purification

Journal: Frontiers in Molecular Neuroscience

Article Title: Connexin30.2: In Vitro Interaction with Connexin36 in HeLa Cells and Expression in AII Amacrine Cells and Intrinsically Photosensitive Ganglion Cells in the Mouse Retina

doi: 10.3389/fnmol.2016.00036

Figure Lengend Snippet: Colocalization (coloc.) of β-galactosidase immunoreactivity with different calcium-binding proteins (CaBP) .

Article Snippet: Calretinin , Guinea pig calretinin, full length amino acid sequence , Goat, polyclonal , 1:500 , Merck-Millipore, Darmstadt, Germany (AB 1550).

Techniques:

Journal: Frontiers in Molecular Neuroscience

Article Title: Connexin30.2: In Vitro Interaction with Connexin36 in HeLa Cells and Expression in AII Amacrine Cells and Intrinsically Photosensitive Ganglion Cells in the Mouse Retina

doi: 10.3389/fnmol.2016.00036

Figure Lengend Snippet: Cx30.2 lacZ -expressing neurons belong to several different populations of amacrine and ganglion cells, including intrinsically photosensitive retinal ganglion cells (ipRGCs). (A–H) Double immunofluorescence stainings for β-gal (A 1 –H 1 ) and calbindin (A 2 ,B 2 ) , calretinin (C 2 ,D 2 ) , parvalbumin (E 2 ,F 2 ) , and melanopsin (G 2 ,H 2 ) in the INL (A/C/E/G) and GCL (B,D,F,H) of Cx30.2 lacZ/lacZ whole-mount retinae. (A 3 –H 3 ) show overlays of the respective single channels. Arrows indicate example cells that express both antigens, double arrows indicate cells that express β-gal but lack the respective marker, and arrowheads indicate cells expressing the marker but lacking β-gal. Insets in (G 1 ) show intensity-enhanced magnifications of the areas marked by dashed squares to verify colocalization with melanopsin. Scale bar: 50 μm.

Article Snippet: Calretinin , Guinea pig calretinin, full length amino acid sequence , Goat, polyclonal , 1:500 , Merck-Millipore, Darmstadt, Germany (AB 1550).

Techniques: Expressing, Immunofluorescence, Marker

Journal: bioRxiv

Article Title: Maturation of Purkinje cell firing properties relies on granule cell neurogenesis

doi: 10.1101/2020.05.20.106732

Figure Lengend Snippet: A. Schematic of an embryonic brain. Inset is the cerebellar anlage. Atoh1 domain (granule cell precursors, pink), Ptf1a domain (Purkinje cell precursors, green), En1 domain (grey). Orientation is the same for all panels unless otherwise indicated. B. Schematic of a sagittal section of a P14 cerebellum. Purkinje cell=green; granule cell=pink. Cerebellar lobules are labelled with Roman numerals . C. Intersectional labelling of En1;Atoh1 domain with tdTomato (pink) shows no overlap with Purkinje cells (Calbindin; green). D. Whole brain images of control and En1 Cre/+ ;Atoh1 fl/- mice showing abnormal gross morphology. E. Sagittal sections of control and En1 Cre/+ ;Atoh1 fl/- hindbrains stained with cresyl violet to visualize cell nuclei. F. and G. Sagittal and coronal sections of P14 cerebella of control ( F ) and En1 Cre/+ ;Atoh1 fl/- ( G ) mice stained with Calbindin (grey). Arrows indicate Purkinje cells that have migrated into the colliculi. F. and G. are presented at the same magnification. H. Higher magnification images of Calbindin staining in control and En1 Cre/+ ;Atoh1 fl/- mice. Images are representative for N=3 brains for each genotype.

Article Snippet: The following primary antibodies were used for the data described in this manuscript: guinea pig (gp)-α-Calbindin (1:1,000; SySy; #214004); rabbit (rb)-α-gamma-aminobutyric acid receptor α6 (GABARα6; 1:500; Millipore Sigma; #AB5610), rb-α-T-box brain protein 2 (Tbr2; 1:500; Abcam; #AB23345), mouse (ms)-α-Calretinin (1:500; Swant; #6B3); ms-α-Neurofilament Heavy (NFH; 1:1,000; Biolegend; #801701); rb-α-Hyperpolarization Activated Cyclic Nucleotide Gated Potassium Channel 1 (HCN1; 1:500; Alomone Lab; #APC-056); goat (gt)-α-RAR-related orphan receptor alpha (RORα; 1:250; Santa Cruz; #F2510); rb-α-parvalbumin (PV; 1:1,000; Swant; #PV25); rb-α-neurogranin (1:500; Millipore Sigma; #AB5620); ms-α-ZebrinII (1:500; kind gift from Dr. Richard Hawkes, University of Calgary, Calgary, Alberta, Canada); rb-α-Vglut1 (1:500; SySy; #135302); rb-α-Vglut2 (1:500; SySy; #135403).

Techniques: Control, Staining

Journal: bioRxiv

Article Title: Maturation of Purkinje cell firing properties relies on granule cell neurogenesis

doi: 10.1101/2020.05.20.106732

Figure Lengend Snippet: A. Schematic of a control Purkinje cell and an En1 Cre/+ ;Atoh1 fl/- Purkinje cell (based on results from B-H ). MF=Mossy Fiber; CF=Climbing fiber. B. Images of Calbindin (green) and Vglut1 (pink) staining. For En1 Cre/+ ;Atoh1 fl/- mice in B, C, and E c=cerebellum, and d=displaced Purkinje cells. Images are representative for N=3 brains for each genotype. C. Images of Calbindin (green) and Vglut2 (pink) staining. D. Schematic of a control Purkinje cell, the En1 Cre/+ ;Atoh1 fl/- ZebrinII staining pattern (see Supp. Figure 2 ) and WGA-Alexa 555 tracing form the spinal cord. E. Representative images of WGA-Alexa 555+ terminals in the cerebellum. Dotted lines represent the border between ZebrinII-positive (cyan) and -negative region (left four images) or between the cerebellum and colliculi (right images). Black and white image shows the pattern of WGA-Alexa 555 positive terminals. F. Representative images of Golgi-Cox-labelled Purkinje cells in control (top row) and En1 Cre/+ ;Atoh1 fl/- brains (bottom row). G. Sholl analysis for dendritic complexity. H. Purkinje cells in En1 Cre/+ ;Atoh1 fl/- mice have shorter and less branched Purkinje cell dendrites (n=30/N=3 for each genotype, each animal is indicated with a differentially oriented triangle). Linear mixed model with genotype as the fixed effect and mouse number as the random effect. *P<0.001 for both distance from soma and branch number. All images were acquired from the cerebellum of P14 mice.

Article Snippet: The following primary antibodies were used for the data described in this manuscript: guinea pig (gp)-α-Calbindin (1:1,000; SySy; #214004); rabbit (rb)-α-gamma-aminobutyric acid receptor α6 (GABARα6; 1:500; Millipore Sigma; #AB5610), rb-α-T-box brain protein 2 (Tbr2; 1:500; Abcam; #AB23345), mouse (ms)-α-Calretinin (1:500; Swant; #6B3); ms-α-Neurofilament Heavy (NFH; 1:1,000; Biolegend; #801701); rb-α-Hyperpolarization Activated Cyclic Nucleotide Gated Potassium Channel 1 (HCN1; 1:500; Alomone Lab; #APC-056); goat (gt)-α-RAR-related orphan receptor alpha (RORα; 1:250; Santa Cruz; #F2510); rb-α-parvalbumin (PV; 1:1,000; Swant; #PV25); rb-α-neurogranin (1:500; Millipore Sigma; #AB5620); ms-α-ZebrinII (1:500; kind gift from Dr. Richard Hawkes, University of Calgary, Calgary, Alberta, Canada); rb-α-Vglut1 (1:500; SySy; #135302); rb-α-Vglut2 (1:500; SySy; #135403).

Techniques: Control, Staining

Journal: Frontiers in Cellular Neuroscience

Article Title: Dentate gyrus network dysfunctions precede the symptomatic phase in a genetic mouse model of seizures

doi: 10.3389/fncel.2013.00138

Figure Lengend Snippet: SynII is highly colocalized with the mossy cell marker calretinin. (A) Representative images of the DG region in brain slices from 6 weeks old WT mice immunostained for Syn I, Syn II, and calretinin (scale bar 50 μm); (B) Representative images of the inner molecular layer area (IML) of the DG immunostained for Syn I (upper panel) or Syn II (lower panel) (scale bar, 50 μm); (C,D) Mean (±s.e.m.) ratios between Syn I/calretinin and Syn II/calretinin signals in the hilus (Hyl, C ) and inner molecular layer (IML, D ) of the DG. * p < 0.05, two-tailed unpaired Student's t -test. (E) Representative images of the DG region in brain slices from 6 weeks old WT (left column), Syn I −/− (middle column) or Syn II −/− (right column) mice immunostained for all synapsin isoforms (pan-Syn antibody) and calretinin. Right panels display the merge of the two signals (scale bar, 50 μm).

Article Snippet: Antibodies used were: Anti-Synapsin 1 (Mouse, 1:500, SYSY #106 001; Rabbit, 1:200 G-177), Anti-Synapsin 2 (Mouse, 1:200, clone 19.21), Anti-panSynapsins (Rabbit, 1:500 G143), Anti-Calretinin (Guinea Pig, 1:1000, SYSY# 214 104).

Techniques: Marker, Two Tailed Test

Journal: bioRxiv

Article Title: Anatomical development of the cerebellothalamic tract in embryonic mice

doi: 10.1101/731968

Figure Lengend Snippet: A) Horizontal view of a maximum intensity projection of a 3DISCO cleared E17.5 mouse brain. No visible change has occurred compared to E16.5 at this level of magnification. Compass in bottom right indicating directions of rostral (R) and caudal (C) sides B-E) Coronal section of an E17.5 mouse brain, with insets. Fibers can be seen in the Th, though they are much less bundled than in the medial pTh. F) Coronal section of an E18.5 mouse brain stained for FoxP2 (in green), Calbindin (Magenta) and RFP (in red) showing an example of delineation of the VM, POm, MD, VB, VL, LP, dLGN, fr, and the mt. G) Zoom in of boxed region in F. In the inset, a single axon with multiple putative boutons is visible. Arrows indicate the putative boutons. H) Binarized version of G after applying the threshold. Inset is a binarized version of the inset in H. Scale bars: A = 100 µm; B and D = 250 µ m; C and E = 100 µ m; F = 200 µm; G = 100 µm, insets = 25 µm. CbT = cerebellothalamic tract; CbX = decussation of the CbT; CN = cerebellar nuclei; dLGN = dorsal lateral geniculate nucleus; fr = fasciculus retroflexus; Hb = Habenula; Hy = hypothalamus; LP = lateral posterior nucleus; MD = mediodorsal nucleus; mt = mamillothalamic tract; Pf = Parafascicular nucleus; POm = posteromdeial nucleus; VB = ventrobasal nucleus; VL = ventrolateral nucleus; VM = ventromedial nucleus.

Article Snippet: For immunofluorescence, we used chicken anti-Calbindin (1:500, Synaptic Systems ), mouse anti-CalbindinD-28 (1:500, Sigma ), chick anti-GFP (1:200, Abcam ), goat anti-FoxP2 (1:500, Santa Cruz ), guinea pig anti-vGluT2 (1:500, MilliPore ), rabbit anti-RFP (1:1000, Rockland ), and mouse anti-NeuN (1:1000, MilliPore ) as primary antibodies.

Techniques: Staining