Journal: eLife

Article Title: Regulation of retinal axon growth by secreted Vax1 homeodomain protein

doi: 10.7554/eLife.02671

Figure Lengend Snippet: ( A ) Cells expressing Vax1 (green) in brain sections (coronal; 16 μm) from E14.5 Vax1 +/+ (top) and Vax1 −/− (bottom) embryos were detected by co-immunostaining for the NPC marker Sox2 (red) and post-mitotic neuronal marker tubulin-βIII (blue), detected with the Tuj1 antibody. The right-most three columns are the magnified images of dotted boxes in the left column image. The results indicate that Vax1 is expressed in a subpopulation of Sox2-positive NPCs (arrowheads) but is not detectable in Tuj1-positive neurons. ( B ) Vax1-expressing cells in the vHT were also compared with RC2-positive radial glia. Arrowheads indicate RC2-positive radial glial cells expressing Vax1. Scale bars: 50 μm. ( C ) The vHT and dorsal neural retina (NR) were isolated from WT ( Vax1 +/+ ) and Vax1- knockout ( Vax1 −/− ) E13.5 mouse embryos and co-cultured in a combinatorial manner for 48 hr. The explants were fixed and immunostained with an anti-NF160 antibody (α-NF160; red); nuclei were counterstained with DAPI (blue). Dotted boxes indicate the area magnified in each inset. Red dotted lines link centers of retinal explants and vHT explants. Scale bars: 500 μm. ( D ) The angular distribution of RGC axons in images was measured by counting pixels containing immunostaining for the axon marker NF160 (axon counts), as described in ‘Materials and methods’, and presented graphically. +, forward direction angle segment; 0, neutral direction angle segments; −, reverse direction angle segment. The values in the bar are averages, error bars denote standard deviations (SDs), and numbers under y -axis labels are the numbers (n) of explants analyzed from three independent experiments. p -values determined by the analysis of variance (ANOVA) are between 0.01 and 0.005. DOI: http://dx.doi.org/10.7554/eLife.02671.003

Article Snippet: Commercially available antibodies against the following proteins were used: mouse anti-Myc (Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-GFP (Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-tubulin β-III (Tuj1; Covance, Princeton, NJ, USA), goat anti-Sox2 (Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-Nestin (RC2; Millipore, Billerica, MA, USA), mouse anti-β-galactosidase (Developmental Studies Hybridoma Bank, DSHB), mouse anti-NF160 (Developmental Studies Hybridoma Bank [DSHB], Iowa City, IA, USA), goat anti-Sdc2 (Santa Cruz Biotechnology, Dallas, TX, USA), goat anti-Sdc3 (Santa Cruz Biotechnology (Dallas, TX, USA), for immunohistochemistry), rabbit anti-Sdc3 (Abcam (UK), for Western blot), rabbit anti-Glp1 (Santa Cruz Biotechnology, Dallas, TX, USA), and rabbit anti-Pax2 (Invitrogen, Carlsbad, CA, USA) antibodies.

Techniques: Expressing, Immunostaining, Marker, Isolation, Knock-Out, Cell Culture

Journal: Frontiers in Cellular Neuroscience

Article Title: Differentiation-Dependent Motility-Responses of Developing Neural Progenitors to Optogenetic Stimulation

doi: 10.3389/fncel.2017.00401

Figure Lengend Snippet: Primary antibodies used.

Article Snippet: Anti- RC2 mouse IgM monoclonal , E14 - E17 rat fetal brain , Developmental Studies Hybridoma Bank; University of Iowa, United States , 1/500.

Techniques: Recombinant, Sequencing

Journal: Frontiers in Cellular Neuroscience

Article Title: Differentiation-Dependent Motility-Responses of Developing Neural Progenitors to Optogenetic Stimulation

doi: 10.3389/fncel.2017.00401

Figure Lengend Snippet: Immunocytochemical characterization of ChR2-eYFP expressing and non-expressing RGl cells. ChR2-eYFP expressing RGl cells were identified by eYFP fluorescence (A,C,E,G,I,K: green ). Both ChR2+ (A–L) and ChR2- cells (M–P) displayed radial glial markers as nestin ( B–D,M,N : red), RC2 ( F–H,O,P : greenish blue) and Pax6 ( J,K : red). Blue color shows DAPI staining of cell nuclei. All pictures were taken at the same magnification. Scale bars are presented on (L,P) .

Article Snippet: Anti- RC2 mouse IgM monoclonal , E14 - E17 rat fetal brain , Developmental Studies Hybridoma Bank; University of Iowa, United States , 1/500.

Techniques: Expressing, Fluorescence, Staining

Journal:

Article Title: Secretagogin is a Ca 2+ -binding protein identifying prospective extended amygdala neurons in the developing mammalian telencephalon

doi: 10.1111/j.1460-9568.2010.07275.x

Figure Lengend Snippet: Generation of anti-secretagogin antibodies and histochemistry

Article Snippet: Tissue autofluorescence in sections from adult mouse and primate brains was quenched by Sudan Black B ( Schnell et al. , 1999 ). table ft1 table-wrap mode="anchored" t5 Protein target Species Dilution Source Cat. No. Fluorophore Reference Brn-1 goat 1:1,000 Santa Cruz SC-6028 Cy2 Keays et al . (2007) Calretinin goat 1:2,000 SWant CG1 Cy5 Schwaller et al . (1993) Calbindin D28k mouse (IgG) 1:2,000 SWant 300 Cy5 Celio (1990) Choline-acetyltransferase goat 1:200 Chemicon/Millipore AB144P Cy2, Cy3 Härtig et al . (1998) Doublecortin goat 1:100 Santa Cruz SC-8066 Cy2, Cy3 Nagatsuka et al . (2003) GABA guinea pig 1:1,000 Chemicon/Millipore AB175 Cy3 McDonald & Pearson (1989) GAD65/67 mouse (IgG) 1:500 Nordic BioSite MSA-225 Cy5 Karlsen et al . (1991) Lhx8 goat 1:100 Santa Cruz SC-2216X Cy3, Cy5 Kitanaka et al . (1998) Nestin mouse (IgG) 1:1,000 Chemicon/Millipore MAB353 Cy5 Bossolasco et al . (2005) PSA-NCAM mouse (IgM) 1:200 Chemicon/Millipore MAB5324 Cy5 Bernier et al . (2002) RC2 mouse (IgM) 1:200 Chemicon/Millipore MAB5740 Cy5 Aguado et al . (2006) Secretagogin goat 1:1,000 R & D Systems AF4878 Cy3 This report β-III-tubulin (TUJ1) mouse (IgG) 1:2,000 Promega G7121 Cy5 Berghuis et al . (2007) Tyrosine hydroxylase mouse (IgG) 1:1,000 R & D Systems MAB1423 Cy2 Arita et al . (2002) Open in a separate window Generation of anti-secretagogin antibodies and histochemistry

Techniques:

Journal: bioRxiv

Article Title: The valine-arginine dipeptide repeat protein encoded by mammalian telomeric RNA appears highly expressed in mitosis and may repress global translation

doi: 10.1101/2024.07.24.604971

Figure Lengend Snippet: (A) E16 mouse embryonic cortical section was labeled with anti-VR (teal) and cortical progenitor-specific RC2 (Yellow) antibodies. VR labels the ventricular zone (VZ) where progenitors proliferate as well as the cortical plate (CP) where new neurons settle. (B-C) VR expression is also evident in the early (E14) and late-stage (P0) cortical ventricular zone. Anti-VR and RC2 antibodies were used at 1:200 dilution. IZ, intermediate zone. Scale bar: 50µm (A), 25µm (B-C).

Article Snippet: The following primary antibodies were used: anti-VR ( ) and RC2 (1:200 dilution; mouse IgM, Developmental Studies Hybridoma Bank, University of Iowa).

Techniques: Labeling, Expressing

Journal: PLoS ONE

Article Title: Low Density Lipoprotein-Receptor Related Protein 1 Is Differentially Expressed by Neuronal and Glial Populations in the Developing and Mature Mouse Central Nervous System

doi: 10.1371/journal.pone.0155878

Figure Lengend Snippet: Coronal sections of the E13.5 (a), E15.5 (c) and E18 (g)mouse brain and transverse sections of the E15.5 spinal cord (e) were immunolabelled to detect radial glia (RC2, green) and LRP1 (red). The nuclear marker Hoechst 33342 was used to label cell nuclei (blue). (b,d,f,h,j) secondary antibody alone controls. All images are single z plane confocal scans. White arrows indicate regions of co-localisation. Scale bars represent 17μm. SC = spinal cord.

Article Snippet: Primary antibodies included rabbit anti-LRP1 (1:500, ab92544, Abcam), goat anti-PDGFRα (1:100, AF1062, R&D Systems), mouse anti-PSANCAM (1:500, MAB5324, Millipore), mouse anti-RC2 (1:100, MAB5740, Millipore), mouse anti-GFAP (1:2000, 556327, BD Pharmigen), guinea-pig anti-Iba1 (1:250, 234004, Synaptic Systems), mouse anti-CC1 (1:200, MABC200 Millipore), mouse anti-NeuN (1:200, MAB377, Millipore), mouse anti-parvalbumin (1:1000, MAB1572, Millipore) and rat anti-GFP (1:2000, Nacalitesque).

Techniques: Marker

Journal: Neural Development

Article Title: Migration, early axonogenesis, and Reelin-dependent layer-forming behavior of early/posterior-born Purkinje cells in the developing mouse lateral cerebellum

doi: 10.1186/1749-8104-5-23

Figure Lengend Snippet: The early/posterior-born Purkinje cells are elongated radially and tangentially in the lateral cerebellum at E12.5 and E13.5 . (A-Q) The morphology and orientation of the E10.5-adenovirally labeled nascent Purkinje cells in E12.5 (A-I) and E13.5 (L, O, Q) cerebella are compared with immunoreactivity for Nestin (J, M) (almost identical staining patterns were obtained by using RC2 (not shown)) and Neurofilament (K, N). dn, prospective deep nuclear neurons; RL, rhombic lip. In (I, L), traces of representative cases of E10.5-born nascent Purkinje cells (including cases 1a, 1b, 2, 3, and 4 in this figure, as well as those shown in Additional file ) are summarized in an illustration of a 'standardized' cerebellar primordium at E12.5 or E13.5. The scale is common in panels (I-N). Note that E10.5-born Purkinje cells seen in an outer zone near the pial surface are tangentially elongated whereas those in deeper regions are more radially elongated. Neurofilament + radial fibers markedly increased between E12.5 and E13.5 and appear to outnumber Nestin + fibers at E13.5. (R) Each tangentially oriented Purkinje cell at E13.5 (Q, R) is polarized with a single thin and long process extended anteriorly (arrowheads) and a thick cytoplasmic part (double blue arrowhead) from which a few thick processes (arrow) are extended posteriorly.

Article Snippet: Frozen sections were treated with the following primary antibodies: anti-Lhx1/5 (Lim1/2; mouse, Hybridoma Bank [4F2]); anti-Corl2 (rabbit) [ ]; anti-Nestin (rabbit, gift from Dr Yasuhiro Tomooka) [ ]; RC2 (mouse IgM, gift from Dr Miyuki Mamamoto) [ ]; anti-Neurofilament (mouse, Hybridoma Bank [2H3]; rabbit, BIOMOL, Plymouth Meeting, PA, USA); anti-GFP (rat, Nakarai Tesque, Inc., Kyoto, Japan; rabbit, MBL, Nagoya, Japan, anti-Tbr1 (rabbit, Abcam, Tokyo, Japan), anti-Reelin (mouse [CR50]) [ , ]; Pax6 (rabbit, Covance, Berkeley, CA, USA); anti-Math1 (Hybridoma Bank); anti-tau (mouse, Chemicon, Billerica, MA, USA); anti-MAP2 (mouse, Sigma, St. Louis, MO, USA); and anti-giantin (rabbit, Covance).

Techniques: Labeling, Staining