Secretory patterns and rates of gonadotropin-releasing hormone, follicle-stimulating hormone, and luteinizing hormone revealed by intensive sampling of pituitary venous blood in the luteal phase mare

Abstract

We used our unique nonsurgical technique for collecting pituitary venous (pit) blood to study GnRH, FSH, and LH secretion patterns in midluteal phase mares. This method does not perturb endocrine function and allows continuous monitoring of GnRH and gonadotropin (Gn) secretion, determination of the amount of GnRH perfusing gonadotropes, and direct measurements of the amounts of Gn secreted. In a total of 80 h of 5-min sampling in four mares, eight Gn peaks occurred; however, more frequent sampling was needed to define secretory events precisely. Therefore, pit blood was collected continuously and split into 30-sec segments in six mares. To ensure a peak during sampling, the opioid antagonist naloxone was given after 4-6 h of sampling to try to replicate a physiological signal for GnRH release. Naloxone induced Gn peaks in jugular blood that were indistinguishable in amplitude from spontaneous peaks. Intensive sampling of pit blood showed that jugular peaks reflected major episodes of GnRH and Gn secretion lasting 30-55 min, which were similar in profile whether naloxone induced or spontaneous and consisted of a train of three to six peaks of diminishing amplitude. Peaks of GnRH and, less often, Gn also occurred outside major episodes. Despite markedly variable size, GnRH peak maxima were correlated with the amount of LH and FSH secreted in concurrent peaks. Likewise, cross-correlation analyses (n = 960 samples/mare) showed close correspondence between patterns of GnRH and secreted FSH and LH. The delay (+/- SEM) between GnRH and Gn maxima was 0.62 +/- 0.18 min for LH and 0.18 +/- 0.22 min for FSH. The majority of GnRH and Gn peaks were concurrent; however, 34.7% of GnRH peaks occurred without Gn peaks. These peaks had a lower amplitude than those with Gn peaks (P < 0.001). For Gn, secretion (i.e. ratio between pit and jugular concentrations, > 1.5) continued at a low level for 40 +/- 9% (LH) or 64 +/- 14% (FSH) of the time between Cluster-defined peaks during the basal period. We conclude that in the luteal phase 1) the predominant mode of GnRH and Gn secretion is as concurrent, large amplitude, prolonged episodes that appeared to be the summation of a train of peaks; and 2) a GnRH dose-Gn response relationship operates endogenously. This along with the synchronicity of secretion patterns of the three hormones suggest that GnRH is the major secretagogue for both LH and FSH.