Yang S P, Pau K Y, Spies H G
Oregon Regional Primate Research Center, Beaverton 97006, USA.
J Mol Endocrinol. 1997 Dec;19(3):311-9. doi: 10.1677/jme.0.0190311.
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.
下丘脑去甲肾上腺素(NE)在性行为控制和促性腺激素分泌中起重要作用。我们之前的研究表明,交配可使雌性而非雄性兔子的下丘脑NE和GnRH释放同时增加。为了研究促黄体生成素(LH)峰诱导过程中去甲肾上腺素能神经元的活动,我们测量了雌性新西兰白兔蓝斑(LC)去甲肾上腺素能细胞中酪氨酸羟化酶(TH,NE合成的限速酶)和NE转运体(NET,NE细胞再摄取的关键蛋白)的mRNA水平。作为对照,也测量了雄性兔子LC-TH和LC-NET mRNA水平的变化。雌性兔子在交配前以及交配后15、30、60、120和240分钟处死(每个时间点n = 6 - 7只);雄性兔子在交配前以及交配后30、60和120分钟处死(每个时间点n = 3只)。将单个脑干切片,取出LC神经元,通过核糖核酸酶保护分析(RPA)对TH和NET mRNA进行定量。在RPA中使用兔特异性TH(330 bp)和NET(503 bp)cDNA作为探针以获得基因特异性信号。还克隆了一种兔管家cDNA(亲环蛋白,158 bp)并用作组织RNA含量的内参。采集躯干血以测定血清LH水平。在雌性兔子中,血清LH水平在交配后15分钟开始升高,在1 - 2小时达到峰值浓度,随后下降。雌性兔子中TH和NET mRNA水平变化的时间间隔与血清LH水平相似。TH和NET mRNA在15分钟时均显著增加(分别为73%和85%),并在2小时内升高(分别为87%和111%)。交配后4小时TH mRNA水平恢复到基础水平,而在整个4小时观察期内NET mRNA值一直升高。相比之下,雄性兔子交配后LH、TH和NET mRNA水平没有变化。雌性兔子LC中TH和NET基因表达增强,这与我们之前关于下丘脑NE释放增加的证明一致,表明NE合成和再摄取的调节是交配诱导的NE/GnRH/LH峰过程的一个组成部分,该过程包括这些神经化学物质释放和/或储存的起始、维持或恢复。