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Corticotropin-Releasing Factor Receptor Expression in the Pituitary of Fetal Sheep after Lesion of the Hypothalamic Paraventricular Nucleus¹

Department of Physiology (D.A.M., M.E.B., T.R.M.), College of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73190; and Laboratory for Pregnancy and Newborn Research (T.J.M.), Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853

Address all correspondence and requests for reprints to: Dean A. Myers, Ph.D., Department of Physiology, College of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73190. E-mail: dean-myers{at}ouhsc.edu

Both the capacity of CRF to release ACTH and the number of binding sites for CRF in the anterior pituitary decline during the final weeks of gestation in fetal sheep. The present study examined regulation of pituitary CRF receptor expression by the hypothalamic paraventricular nucleus (PVN) during late gestation in fetal sheep.

Bilateral radiofrequency lesions of the PVN (PVN-Lx; n = 4) or sham lesions (SHAM; n = 5) were performed in fetal sheep at 118–122 days of gestational age (dGA). Pituitary glands from PVN-Lx and SHAM fetuses were collected at 139–142 dGA (term, approximately 148 dGA). Dual-label in situ hybridization was performed using a digoxigenin-labeled ovine POMC complementary RNA, together with a ^35S-labeled ovine CRF type I (CRF₁) receptor complementary RNA, to localize and quantify CRF₁ receptor mRNA in POMC-hybridizing cells. Binding of [¹²⁵I]-ovine CRF was also examined in the fetal pituitary of both PVN-Lx and SHAM fetuses using in situ autoradiography.

The hybridization signal for the CRF₁ receptor mRNA was primarily restricted to POMC-expressing cells in the anterior pituitary of both PVN-Lx and SHAM fetuses; no hybridization signal for the CRF₁ receptor was observed in the neurointermediate lobe (NIL) in either group. The hybridization signal for CRF₁ receptor mRNA in anterior pituitary corticotropes of PVN-Lx fetuses was significantly lower in both the inferior and superior regions of the anterior pituitary, compared with SHAM fetuses (P < 0.05). In the inferior region of the anterior pituitary, the percentage of POMC-hybridizing cells containing CRF₁ receptor hybridization signal was significantly greater in PVN-Lx (90 ± 7%; mean + SEM), compared with SHAM (67 ± 6%; P < 0.05) fetuses. No differences in the percentage of POMC cells containing CRF₁ receptor hybridization signal were observed in the superior region of the anterior pituitary between PVN-Lx (89 ± 8%) and SHAM (87 ± 9%). Binding of [¹²⁵I]-ovine CRF (oCRF) was significantly greater in anterior pituitaries of PVN-Lx (140 ± 19 mean arbitrary densitometry U ± SEM), compared with SHAM (73 ± 23; P < 0.05) fetuses. For both PVN-Lx and SHAM fetuses, there were no differences within group in [¹²⁵I]-oCRF binding between the inferior and superior regions of the anterior pituitary. A weak, but significant (P < 0.05), autoradiographic signal for [¹²⁵I]-oCRF binding was observed in the NIL of both SHAM and PVN-Lx fetal sheep. The level of [¹²⁵I]-oCRF binding was significantly lower in the NIL, compared with anterior pituitary, for both SHAM (P < 0.01) and PVN-Lx fetuses. There were no differences in [¹²⁵I]-oCRF binding in the NIL between SHAM and PVN-Lx fetal sheep. Our findings support a role for the PVN in regulating anterior pituitary CRF₁ receptor expression in the late-gestation sheep fetus.

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