ZFIN ID: ZDB-FIG-111102-5
Gutnick et al., 2011 - The hypothalamic neuropeptide oxytocin is required for formation of the neurovascular interface of the pituitary. Developmental Cell   21(4):642-654 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Condition:
Anatomical Terms:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage: Protruding-mouth

Fig. 5

Abnormal Development of Pituitary Vasculature in the Absence of HN Innervation

(A and B) Immunostaining of oxytocin (Oxtl) in wild-type (A) and the endothelial-deficient mutant cloche (- / -) (B) mutants, which lack all blood vessels.

(C–L) Genetic cell ablation of Otp+ neurons of the neurosecretory preoptic nucleus (NPO). (C) Quantitative analysis of the vascular phenotype caused by HN neuronal ablation (see text). Using the Gal4/UAS system (otpb:Gal4;UAS:NTR-Cherry) to drive expression of a Nitroreductase-Cherry (NTR-Cherry) fusion protein only in Otp+ neurons make these cells and their projections visible (red) and susceptible to the prodrug metronidazole. This transgenic fish was crossed to the vascular reporter line, vegfr2:EGFP, to generate a triple transgenic line (D–F and J–L). Double transgenic line (otpb:Gal4;vegfr2:EGFP, G–I) expressing vascular EGFP but not the NTR-Cherry protein served as a control for nonspecific drug toxicity. The presence of Cherry+ neurons (D, G, J), EGFP+ blood vessels (E, H, and K) and oxytocin immunoreactivity (F, I, and L) were monitored in three experimental conditions; the respective genotypes as well as the application of the metronidazole killing agent are indicated below. Filled and empty arrowheads indicate normal and abnormal blood vessels, respectively.

See also Figure S3.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP c264Tg; s843Tg; zc57Tg standard conditions Protruding-mouth hypophysis vascular endothelium IHC
c264Tg; s843Tg; zc57Tg chemical treatment: metronidazole Protruding-mouth hypophysis vascular endothelium IHC
s843Tg; zc57Tg chemical treatment: metronidazole Protruding-mouth hypophysis vascular endothelium IHC
mCherry c264Tg; s843Tg; zc57Tg standard conditions Protruding-mouth neurosecretory neuron neuronal cell body IHC
Protruding-mouth preoptic area neurosecretory neuron IHC
c264Tg; s843Tg; zc57Tg chemical treatment: metronidazole Protruding-mouth neurosecretory neuron neuronal cell body IHC
Protruding-mouth preoptic area neurosecretory neuron IHC
oxt Ab2-oxt WT standard conditions Protruding-mouth neurohypophysis axon terminus IHC
Ab2-oxt Protruding-mouth neurosecretory neuron axon terminus IHC
Ab2-oxt npas4lDf(Chr13:lycat)m39/m39 standard conditions Protruding-mouth neurohypophysis axon terminus IHC
Ab2-oxt Protruding-mouth neurosecretory neuron axon terminus IHC
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
c264Tg; s843Tg; zc57Tg standard conditions Protruding-mouth neurohypophysis has normal numbers of parts of type neurosecretory neuron axon terminus, normal
Protruding-mouth neurohypophysis vasculature morphology, normal
Protruding-mouth preoptic area has normal numbers of parts of type neurosecretory neuron, normal
c264Tg; s843Tg; zc57Tg chemical treatment: metronidazole Protruding-mouth neurohypophysis lacks all parts of type neurosecretory neuron axon terminus, abnormal
Protruding-mouth neurohypophysis vasculature malformed, abnormal
Protruding-mouth positive regulation of angiogenesis decreased occurrence, abnormal
Protruding-mouth preoptic area lacks parts or has fewer parts of type neurosecretory neuron, abnormal
npas4lDf(Chr13:lycat)m39/m39 standard conditions Protruding-mouth neurohypophysis has normal numbers of parts of type neurosecretory neuron axon terminus, normal
Protruding-mouth neurohypophysis lacks all parts of type blood vasculature, abnormal
s843Tg; zc57Tg chemical treatment: metronidazole Protruding-mouth neurohypophysis has normal numbers of parts of type neurosecretory neuron axon terminus, normal
Protruding-mouth neurohypophysis vasculature morphology, normal
Protruding-mouth preoptic area has normal numbers of parts of type neurosecretory neuron, normal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Cell, 21(4), Gutnick, A., Blechman, J., Kaslin, J., Herwig, L., Belting, H.G., Affolter, M., Bonkowsky, J.L., and Levkowitz, G., The hypothalamic neuropeptide oxytocin is required for formation of the neurovascular interface of the pituitary, 642-654, Copyright (2011) with permission from Elsevier. Full text @ Dev. Cell