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Gonadotropin-Releasing Hormone Messenger Ribonucleic Acid Expression Changes before the Onset of the Estradiol-Induced Luteinizing Hormone Surge in the Ewe¹

Laboratory of Neuroendocrinology, The Babraham Institute, Babraham, Cambridge, United Kingdom CB2 4AT

Address all correspondence and requests for reprints to: Dr. J. E. Robinson, Department of Neurobiology, The Babraham Institute, Babraham, Cambridge, United Kingdom CB2 4AT. E-mail: jane.robinson{at}bbsrc.ac.uk

The preovulatory LH surge in the ewe is stimulated by the massive and sustained release of GnRH into the pituitary portal vessels. This study has examined the temporal relationship between changes in LH secretion and GnRH messenger RNA (mRNA) expression at the time of the estradiol-induced LH surge. Ovariectomized Clun Forest ewes were treated with exogenous progesterone and estradiol (E) to mimic estrous cycle concentrations of these gonadal steroids and to induce the LH surge. Ewes were killed at five time points relative to the time of onset of the LH surge: pre-E, before E insertion (n = 6); presurge, after E insertion and 8–10 h before surge onset (n = 5); ascending limb, 2–6 h after surge onset (n = 5); midpeak, 9–12 h after surge onset (n = 5); and postsurge, 21–27 h after surge onset (n = 5). Control animals (n = 5/group), which received no E, were killed at identical time intervals alongside the E-treated ewes. Coronal sections containing the diagonal band of Broca through to the anterior hypothalamus were processed for cellular in situ hybridization using an ³⁵S-labeled oligonucleotide probe complementary to ovine GnRH. No changes were found in the number of GnRH mRNA-expressing cells detected in the rostral preoptic area or the medial septum in either gonadal steroid-treated or control ewes. In contrast, cellular GnRH mRNA expression (as assessed by silver grain density) decreased significantly (P < 0.05) between presurge and ascending limb groups within both the rostral preoptic area (0.64 ± 0.06 vs. 0.43 ± 0.05 silver grain density/µm²) and medial septum cells (1.08 ± 0.09 vs. 0.77 ± 0.07). No significant changes were detected in control ewes. These results show that the estradiol-induced LH surge in the ewe is associated with a decrease in GnRH mRNA expression that occurs in advance of the onset of the GnRH surge. This suggests that neural mechanisms controlling GnRH biosynthesis may be distinct from those regulating GnRH secretion.

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