|
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.
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