NEUROCHEMICALS IN HYPOTHALAMIC SIGNALING AFTER COITUS

描述：(改编自申请者摘要)性交导致 新西兰大白兔神经体液事件的串联 包括去甲肾上腺素(NE)分泌增加，促性腺激素释放 激素(GnRH)、黄体生成素(LH)、卵巢类固醇，最后是 释放卵子。合成去甲肾上腺素的脑干神经元是一个可能的成分 酪氨酸羟化酶(TH)表达增强的信号通路 NE转运体(Net)mRNAs在性交后15分钟出现在基因座 蓝斑(LC)，同时下丘脑内注射任一种哌唑嗪(α1 肾上腺素能受体阻滞剂)或地西帕明(Net阻滞剂)可改变GnRH/LH峰。 P.I.提出了三个新的目标来扩大对这一点的理解 神经化学过程。目标1将识别神经递质和神经元 将性交信号传递到LC和/或下丘脑的回路。 神经肽Y(NPY)基因在兔LC中未见表达； 然而，TH、NPY和/或Net mRNAs在 巨细胞旁核(NPGi)或舌下前叶核(NPH)--两种 支配LC的脑干核团。私家侦探假设阴道 信号到达nPGi和/或NPH，在那里它们兴奋中间神经元(即，NPY， 乙酰胆碱[ACH])，可兴奋LC或 直接投射到下丘脑细胞。目标1a将通过现场确认 杂交(ISH)CFos标记脑干神经元早期诊断的证据 性交后激活的细胞。目标1b将识别TH、Net、NPY、 神经肽Y受体、乙酰胆碱酯酶(AChE)和谷氨酸脱羧酶(GAD) Lc、nPGi、NPH用ISH法测定。AIM 1c将定义特定的细胞类型 交配后即为上述双重染色，又表达CFOS蛋白 (ISH用于mRNAs，免疫细胞化学用于CFOS蛋白)。Aim 2将检查 ACH和GABA对性交后GnRH/LH的影响 通过脑干/下丘脑连接的动作电涌。私家侦探会 给予特定的ACH阻滞剂(α-芋螺毒素)和GABAA激动剂 (麝香醇)局部进入下丘脑或第四脑室 在脑干相关神经元附近。交配诱导的促性腺激素释放激素/黄体生成素分泌 脑干TH/Net/NPY mRNAs将在用药和不用药的兔身上进行测量 治疗。AIM 3将通过显微注射以下两种方法之一来靶向脑干基因座 反义寡核苷酸或电损伤。目标3a将破译如果 LC块内注射TH或Net反义寡核苷酸 性交诱导的NE/GnRH释放。Aim 3b将确定电气设备是否 NPGi和/或NPH的损害阻止性交后表达 LC CFOS蛋白、TH/Net mRNAs或黄体生成素激增。这些研究可能会提供 关于神经学相关不孕症的信息。

DESCRIPTION: (adapted from the applicants abstract) Coitus induces a concatenation of neurohumoral events in New Zealand White rabbits that includes increased secretion of norepinephrine (NE), gonadotropin-releasing hormone (GnRH), luteinizing hormone (LH), ovarian steroids, and finally the release of ova. Brainstem neurons that synthesize NE are a likely component of the signaling pathway as enhanced expression of tyrosine hydroxylase (TH) and NE transporter (NET) mRNAs occurs by 15-min postcoitus in locus coeruleus (LC), while intrahypothalamic infusion of either prazosin (alpha1 adrenergic blocker) or desipramine (NET blocker) alters the GnRH/LH surge. The P.I. proposes three new aims to extend the understanding of this neurochemical process. Aim 1 would identify neurotransmitters and neuronal circuits that relay coital signals to the LC and/or hypothalamus. Neuropeptide Y (NPY) mRNA expression was not observed in the rabbit LC; however, TH, NPY and/or NET mRNAs were co-localized in the paragigantocellularis (nPGi) or praepositus hypoglossi (nPH) nuclei--two brainstem nuclei that innervate the LC. The P.I. hypothesizes that vaginal signals reach the nPGi and/or nPH where they excite interneurons (i.e., NPY, acetylcholine [ACH]) that either stimulate the NE/NET neurons in LC or project directly to hypothalamic cells. Aim 1a will identify by in situ hybridization (ISH) cfos-labeled brainstem neurons for evidence of early postcoital activated cells. Aim 1b will identify cells for TH, NET, NPY, NPY receptors, ACH esterase (Ache) and glutamic acid decarboxylase (GAD) in the LC, nPGi and nPH by ISH. Aim 1c will define the specific cell types after coitus that dual stain for the above and also express cFos protein (ISH for mRNAs, immunocytochemistry for cFos protein). Aim 2 will examine if ACH and gamma aminobutyric acid (GABA) affect the postcoital GnRH/LH surge by action through brainstem/hypothalamic connections. The P.I. will administer a specific ACH blocker ( alpha-conotoxin) and GABAA agonist (muscimol) either locally into the hypothalamus or into the 4th ventricle near brainstem-relevant neurons. Mating-induced GnRH/LH secretion and brainstem TH/NET/NPY mRNAs will be measured in rabbits with and without drug treatment. Aim 3 will target brainstem loci by either microinjection of antisense oligonucleotides or electrical lesioning. Aim 3a will decipher if administration of TH or NET antisense oligonucleotides into the LC blocks the coital-induced NE/GnRH release. Aim 3b will determine if electrical lesions of the nPGi and/or nPH prevent either the postcoital expression of LC cFos protein, TH/NET mRNAs or the LH surge. These studies may provide information on neurologically related infertility.