Coital activation of the hypothalamo-hypophyseal ovarian axis (HOA) is well documented in rabbits, but coital excitation of the hypothalamo-hypophyseal testicular axis (HTA) is less well described. We and others have postulated that the response of the HOA to coitus, as characterized by a dramatic release of hypothalamic GnRH, may be mediated by an increase in norepinephrine (NE) neuronal activity. Herein, we studied selective HOA and HTA responses in New Zealand White rabbits before, during, and after coitus. Firstly, we determined the effects of microdialysis (mu D) and blood-sampling methods on plasma LH and testosterone (T) patterns in male rabbits during sexual performance. Subsequently, we compared the patterns of release in GnRH and norepinephrine (NE) from the arcuate nucleus-median eminence (AME) at 10-min intervals with changes in plasma LH levels in copulating male and female rabbits. Lastly, in 2.5-min AME mu D samples from females immediately after coitus, we measured NE and GnRH concentrations to determine whether NE release precedes that of GnRH. Tethered, freely moving rabbits were exposed to their partners for 10 min at the end of the third (10-min sampling for 5-7 h) or second (2.5-min sampling for 4 h) hour. Data from individuals that did not mate during the 10-min of pairing in the 3- to 7-h sampling trials were included as a control group (sham-mated). The results showed no changes (P> 0.05) in plasma LH and T in either mated (LH: pre, 0.13 +/- 0.08 ng/ml; post, 0.15 +/- 0.03 ng/ml; T: pre, 2.39 +/- 1.20 ng/ml; post, 0.85 +/- 0.26 ng/ml) or sham-mated males (LH: pre, 0.21 +/- 0.08; post, 0.25 +/- 0.10 ng/ml; T: pre, 1.46 +/- 0.51 ng/ml; post, 1.40 +/- 0.38 ng/ml). Likewise, coitus did not alter patterns of AME-NE (pre, 0.47 +/- 0.25; post, 0.56 +/- 0.25 ng/ml) and GnRH (pre, 0.61 +/- 0.45; post, 0.74 +/- 0.32 pg/ml) in mated or sham-mated males. The constant HTA activity during coitus in males appears to be independent of experimental manipulation per se because LH and T levels between mu D (0.18 +/- 0.05 and 1.72 +/- 0.85 ng/ml, respectively) and non-mu D (0.16 +/- 0.05 and 1.52 +/- 0.36 ng/ml, respectively) rabbits were not different (P > 0.05). In contrast to males, females displayed unambiguous and simultaneous increases in NE (P < 0.05) and GnRH (P < 0.01) release from the AME within 10-20 min after coitus; these elevated concentrations in mu D samples lasted for 3-4 h. Microdialysis NE levels averaged 0.02 +/- 0.01 ng/ml before mating, whereas postcoital values averaged 0.09 +/- 0.01 ng/ml. GnRH levels were 1.04 +/- 0.56 and 11.78 +/- 5.06 pg/ml before and after coitus, respectively. Concomitant increases in plasma LH levels were also observed after coitus in these female rabbits. Moreover, measurements of NE and GnRH in 2.5-min mu D samples revealed that the postcoital increase in NE preceded that in GnRH by 2.5-7.5 min (P < 0.05). The results suggest that neuroendocrine circuits in the two sexes of the New Zealand White rabbit respond differently to genital stimulation. In male rabbit...

The central noradrenergic system has a major regulatory role on gonadotropin-releasing hormone/luteinizing hormone (GnRH/LH) secretion in rabbits. Exogenous administration of norepinephrine (NE) alters GnRH/LH release in a sex steroid-dependent manner, i.e. NE stimulates GnRH/LH release in oestrogen-primed ovariectomized (OVX) animals but not in non-primed individuals. To investigate how gonadal steroids influence noradrenergic neuronal activities in the locus coeruleus (LC), mRNA levels of tyrosine hydroxylase (TH) and NE transporter (NET), two key factors regulating NE synthesis and uptake, were compared 3 weeks after gonadectomy (GDX). Intact male (n = 5) and female (n = 6) New Zealand White rabbits were sacrificed along with castrated males (n = 4) and OVX females (n = 5). The brainstem from each individual was sectioned and the LC was punched for detection of TH and NET mRNA levels using the ribonuclease protection assay (RPA). Trunk blood was collected to determine immunoactive serum LH values. Levels of LH were elevated in both males and females after GDX. Luteinizing hormone concentrations averaged 0.10 +/- 0.05 ng/ml in intact males vs 1.64 +/- 0.31 ng/ml in castrated males (P < 0.01) and 0.30 +/- 0.08 ng/ml in intact vs 9.80 +/- 3.50 ng/ml in OVX females (P < 0.05), respectively. Removal of the gonads also increased TH mRNA levels in the LC in both males and females. In intact males, TH mRNA levels were 0.796 +/- 0.181 pg/microgram total RNA, whereas in castrates mRNA levels averaged 1.667 +/- 0.345 pg/microgram total RNA (P < 0.05). In intact females, TH mRNA levels were 0.617 +/- 0.054 pg/microgram total RNA while the OVX group averaged 1.084 +/- 0.202 pg/microgram total RNA (P < 0.05). Similar increases in NET mRNA were noted after GDX in both sexes. In males, NET mRNA levels were 1.461 +/- 0.401 pg/microgram total RNA in intacts vs 3.666 +/- 0.649 pg/microgram total RNA in castrates (P < 0.05). In females, NET mRNA levels averaged 1.336 +/- 0.212 pg/microgram total RNA and 3.297 +/- 0.835 pg/microgram total RNA in the intact and OVX groups, respectively (P < 0.05). The data indicate that GDX enhances gene expression of both TH and NET. The results support the hypothesis that the feedback regulation of sex steroids on LH secretion in rabbits of both sexes involves transcriptional/translational processes of at least TH and NET in brainstem NE cells.

Hypothalamic norepinephrine (NE) plays an important role in the control of sexual behavior and in the secretion of gonadotropin. Our previous study showed that coitus induced simultaneous increases in hypothalamic NE and GnRH releases in female but not in male rabbits. To investigate the activities in noradrenergic neurons during the coitus-induced process of an LH surge, we measured tyrosine hydroxylase (TH, the rate-limiting enzyme in NE synthesis) and NE transporter (NET, a key protein for NE cellular reuptake) mRNA levels in locus coeruleus (LC) noradrenergic cells in female New Zealand White rabbits. Changes in LC-TH and LC-NET mRNA levels were also measured in males as controls. Female rabbits were killed before coitus and at 15, 30, 60, 120, and 240 min after coitus (n=6-7/time point); males were killed before and at 30, 60, and 120 min after coitus (n=3/time). Individual brainstems were sectioned, the LC neurons punched, and TH and NET mRNAs were quantified by ribonuclease protection assay (RPA). Rabbit-specific TH (330 bp) and NET (503 bp) cDNAs were used as probes in the RPA for gene-specific signals. A rabbit 'house-keeping' cDNA (cyclophilin, 158 bp) was also cloned and used as an internal marker for tissue RNA content. Trunk blood was collected to determine serum LH levels. In female rabbits, serum LH levels rose by 15 min after coitus, reached peak concentrations at 1-2 h, and declined thereafter. The time interval for changes in TH and NET mRNA levels in females was similar to that in serum LH levels. Both TH and NET mRNAs increased significantly by 15 min (73% and 85% respectively) and were elevated for 2 h (87% and 111% respectively). TH mRNA levels returned to basal levels by 4 h after coitus, whereas NET mRNA values were elevated throughout the 4 h of observation. In contrast, LH, TH and NET mRNA levels did not change after coitus in males. The enhanced gene expression of both TH and NET in the LC in females, in accord with our previous demonstration of increased hypothalamic NE release, suggests that regulation of NE synthesis and reuptake is an integral part of the coitus-induced NE/GnRH/LH surge process that includes the initiation, sustenance or recovery of the release and/or storage of these neurochemicals.