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ZFIN ID: ZDB-FIG-111102-6
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:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Protruding-mouth

Fig. 6

Oxytocin and Its Receptor Are Necessary for Pituitary Vascularization

(A–D) High-resolution confocal images of the head vasculature of 3-day-old embryos following targeted knockdown of the gene encoding for oxytocin (oxtl-MO) or oxytocin receptor (oxtlr-MO) using antisense morpholino oligonucliotides, leads to severe impairment of neurohypophyseal vasculature, but does not impede development of neighboring vessels. A schematic representation of the corresponding phenotype with arterial (red) and venous (blue) color-coding is shown in (A2)–(C2).

(D and E) By crossing nuclear- and membrane-tagged fluorescent vascular reporter lines, we are able to count the number of vascular endothelial cells in each vessel. A dot-plot histogram showing the x,y positions of EGFP+;Cherry+ endothelial cells nuclei of the hypophyseal vessels as well as the adjacent internal carotid arteries (CrDI, between the PICA and the PLA) is shown in (D2) and (E2). Individual cells belonging to the hypophyseal vessels (red) and CrDI (gray) are color coded. The oxtlr-MO-injected embryos (E) display a markedly reduced number of endothelial cells in the hypophyseal vessel but no change in the nearby internal carotid artery.

(F) Bar histogram showing EGFP+;Cherry+ cell counts in the internal carotid arteries and hypophyseal vessels in control and following oxtlr knockdown. *p < 0.01, n = 14.

(G) RT-PCR analysis of vegfr2:Cherry-positive and -negative FACS sorted endothelial cells demonstrating vascular expression of the oxytocin receptor (oxtlr).

CaDI, caudal division of internal carotid; CrDI, cranial division of the internal carotid artery; MO, morpholino antisense oligonucleotide; OA, optic artery; PICA, primitive internal carotid artery; PLA, palatocerebral artery; PLV, palatocerebral vein. The asterisk (*) demarcates the location of the neurohypophysis. See also Figure S4.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
EGFP s896Tg; y7Tg standard conditions Protruding-mouth hypophysis vascular endothelium IHC
Protruding-mouth vascular endothelium nucleus IHC
s896Tg; y7Tg + MO1-oxtr standard conditions Protruding-mouth hypophysis vascular endothelium IHC
Protruding-mouth vascular endothelium nucleus IHC
kdrl s896Tg standard conditions Protruding-mouth blood vessel endothelial cell RTPCR
mCherry s896Tg standard conditions Protruding-mouth hypophysis vascular endothelium IHC
s896Tg + MO1-oxtr standard conditions Protruding-mouth hypophysis vascular endothelium IHC
s896Tg + MO2-oxt standard conditions Protruding-mouth hypophysis vascular endothelium IHC
s896Tg; y7Tg standard conditions Protruding-mouth hypophysis vascular endothelium IHC
s896Tg; y7Tg + MO1-oxtr standard conditions Protruding-mouth hypophysis vascular endothelium IHC
otpb s896Tg standard conditions Protruding-mouth Not Detected blood vessel endothelial cell RTPCR
oxtr s896Tg standard conditions Protruding-mouth blood vessel endothelial cell RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
s896Tg + MO1-oxtr standard conditions Protruding-mouth angiogenesis decreased occurrence, abnormal
Protruding-mouth neurohypophysis vasculature malformed, abnormal
s896Tg + MO2-oxt standard conditions Protruding-mouth angiogenesis decreased occurrence, abnormal
Protruding-mouth neurohypophysis vasculature malformed, abnormal
s896Tg; y7Tg + MO1-oxtr standard conditions Protruding-mouth hypophysis vasculature has fewer parts of type blood vessel endothelial cell, abnormal
Protruding-mouth internal carotid artery has normal numbers of parts of type blood vessel endothelial cell, normal
Protruding-mouth neurohypophysis vasculature hypoplastic, abnormal
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