ZFIN ID: ZDB-FIG-130907-8
Alunni et al., 2013 - Notch3 signaling gates cell cycle entry and limits neural stem cell amplification in the adult pallium. Development (Cambridge, England)   140(16):3335-47 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Knockdown Reagent:
Anatomical Term:
Stage: Adult
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Adult

Fig. 8

Notch1b is required for the maintenance of progenitor division and fate. (A) Schematic of the notch1b-MO knock-down experiment: brains are analyzed 2 and 5 days after electroporation of fluorescein-labeled notch1b-MO or control-MO in pallial ventricular cells. (B,C) Analysis of the proliferation status (anti-MCM5) (blue) of electroporated (fluorescein-positive) (green) RG (anti-GS) (magenta), assessed by immunocytochemistry 5 days after electroporation. Arrows indicate proliferating RG, usually notch1b-MO-negative. (D) Proportion of MCM5-positive, GS-positive cells within the MO-targeted population, 2 days (P=0.05) and 5 days (**P<0.001) after electroporation (n=3 brains for each condition). (E) Schematic of the fate analysis in notch1b-MO knock-down experiments: a BrdU pulse is applied 2 days after electroporation of fluorescein-labeled notch1b-MO or control-MO in pallial ventricular cells. Brains are analyzed immediately or after a 3-day chase. (F,G) Analysis of BrdU labeling (white) in electroporated (fluorescein-positive) (green, arrows) RG (anti-GS) (magenta), assessed by immunocytochemistry after a 3-day chase. Arrows indicate fluorescein-labeled BrdU-positive cells in control-MO (glial cells F) and in notch1b-MO (G). (H) Proportion of BrdU-positive, GS-positive cells within the MO-targeted population, 2 days (P=0.05) and 5 days (**P<0.001) after electroporation (n=3 brains for each condition). (I) Proportion of type I (GS positive, MCM5 negative), type II (GS positive, MCM5 positive), type III (GS negative, MCM5 positive) and non-progenitor cells (GS negative, MCM5 negative) [presumably neurons, which virtually constitute the only non-progenitor cell type generated from the pallial GZ (Chapouton et al., 2010; Rothenaigner et al., 2011)] within the BrdU-positive MO-targeted population 5 days after electroporation (type II, neurons: **P<0.001) (n=3 brains for each condition). Scale bars: 10 μm. Confocal projection images from four optical planes, each 0.5 μm thick.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
mcm5 AB control Adult dorsal telencephalon radial glial cell IHC
AB + MO2-notch1b standard conditions Adult dorsal telencephalon radial glial cell IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-glul IHC AB control Adult dorsal telencephalon radial glial cell
IHC AB + MO2-notch1b standard conditions Adult dorsal telencephalon radial glial cell
Phenotype Details
Fish Conditions Stage Phenotype
AB + MO2-notch1b standard conditions Adult dorsal telencephalon radial glial cell cellular potency, abnormal
Adult dorsal telencephalon ventricular zone has extra parts of type neuron, abnormal
Adult neuronal stem cell population maintenance decreased occurrence, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development