NEUROCHEMICALS IN HYPOTHALAMIC SIGNALING AFTER COITUS

描述：（改编自申请人摘要）性交诱导 新西兰白色兔中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 激素（GnRH），促黄体生成激素（LH），卵巢类固醇，最后是 释放卵子 合成NE的脑干神经元可能是 酪氨酸羟化酶（TH）的表达增强 和NE转运体（NET）mRNA在性交后15分钟出现在位点 蓝核（LC），而下丘脑内灌注哌唑嗪（α 1 肾上腺素能阻滞剂）或地昔帕明（NET阻滞剂）改变GnRH/LH峰。 私家侦探提出了三个新的目标，以扩大对这一点的理解， 神经化学过程。 目标1将确定神经递质和神经元 将性交信号传递到LC和/或下丘脑的回路。 兔LC未见神经肽Y（NPY）mRNA表达; 然而，TH、NPY和/或NET mRNA共定位于 旁巨细胞核（nPGi）或舌下前置核（nPH）-两个 支配LC的脑干核团。 私家侦探假设阴道 信号到达nPG 1和/或nPH，在那里它们激发中间神经元（即，NPY， 乙酰胆碱[ACH]）刺激LC中的NE/NET神经元， 直接投射到下丘脑细胞。 目标1a将通过现场识别 杂交（ISH）cfos标记的脑干神经元的早期证据 性交后激活的细胞 目标1b将识别TH，NET，NPY， NPY受体、ACH酯酶（Ache）和谷氨酸脱羧酶（GAD）在 ISH测定LC、nPGi和nPH。 目标1c将定义特定的细胞类型 交配后，上述双重染色还表达cFos蛋白 (ISH对于mRNA，免疫细胞化学对于cFos蛋白）。 目标2将检查 如果ACH和γ-氨基丁酸（GABA）影响性交后GnRH/LH 通过脑干/下丘脑连接的动作而激增。 私家侦探将 给予特定ACH阻断剂（α-芋螺毒素）和GABAA激动剂 （蝇蕈醇）局部进入下丘脑或第四脑室 脑干相关神经元附近 交配诱导的GnRH/LH分泌和 将在有和没有药物的兔中测量脑干TH/NET/NPY mRNA 治疗 Aim 3将通过显微注射 反义寡核苷酸或电损伤。 目标3a将破译，如果 将TH或NET反义寡核苷酸施用到LC块中 性交诱导的NE/GnRH释放。 目标3b将确定电气 nPGi和/或nPH的病变阻止了 LC cFos蛋白、TH/NET mRNA或LH峰。 这些研究可以提供 与神经系统相关的不孕症的信息。

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.