Intermittent Hypoxia-Induced Hypertension: Roles of Angiotensin and Chloride Transport in the Lamina Terminalis.

摘要-项目2 我们的工作假设是，基于我们的出版物和初步数据，中位数 视前核(MnPO)通过基因活性依赖性改变参与CIH高血压的发病 FosB介导的表达，由循环血管紧张素II(Ang II)通过 穹隆下器(SFO)。MnPO通过投射到脑区影响交感神经活动 下丘脑室旁核。我们已经在MnPO中发现了几个可能的FosB靶基因 这可能导致包括血管紧张素转换酶1(ACE1)在内的CIH高血压，并且基于我们的 初步资料为血管紧张素受体1a型(AT1aR)。这些FosB靶基因可能共同驱动 MnPO可能通过与钠钾离子相互作用在CIH高血压中的作用 联合转运体1(NKCC1)。 具体目的1：确定MnPO中ACE1在CIH介导的高血压中的作用。 假设：血管紧张素转换酶1在MnPO中的表达增加是CIH高血压所必需的 特异目的2：探讨血管紧张素Ⅱ受体AT1aR在脑出血高血压中的作用。 假设：血管紧张素转换酶AT1aR的表达增加有助于在脑出血期间增强MnPO的激活。 具体目的3：确定NKCC1在CIH高血压患者MnPO中的作用。 假设：NKCC1表达和活性增加可降低GABA对MnPO的抑制作用 导致在CIH期间产生更大的交感神经兴奋。 这些假说将用综合的方法结合整个动物实验进行检验。 利用病毒介导的shRNA敲除、体内交感神经和体外单单位记录 膜片钳记录，qRT-PCR激光捕获显微切割。将使用光遗传方法 使用谷氨酸特异性启动子检测SFO-MnPO通路在CIH高血压中的作用 表达神经元。

Summary – Project 2 Our working hypothesis, which is based on our publications and our preliminary data, is that the median preoptic nucleus (MnPO) contributes to CIH hypertension due to activity dependent changes in gene expression mediated by FosB that are driven by circulating angiotensin II (ANG II) working through the subfornical organ (SFO). The MnPO influences sympathetic nerve activity through its projection to the paraventricular nucleus of the hypothalamus. We have identified several putative FosB target genes in MnPO that may contribute to CIH hypertension including angiotensin converting enzyme 1 (ACE1), and based on our preliminary data the angiotensin receptor type 1a (AT1aR). Together these FosB target genes may drive the MnPO contribution to CIH hypertension through their possible interaction with the sodium-potassium-chloride co-transporter 1 (NKCC1). Specific Aim 1: to determine the contribution of ACE1 in the MnPO to CIH mediated hypertension. Hypothesis: Increased expression of ACE1 in the MnPO is necessary for CIH hypertension Specific Aim 2: to test the role of increased AT1aR expression in the MnPO in CIH hypertension. Hypothesis: Increased AT1aR expression contributes to enhanced activation of the MnPO during CIH. Specific Aim 3: to determine the role of NKCC1 in the MnPO in CIH hypertension. Hypothesis: Increased expression and activity of NKCC1 reduces the inhibitory effects of GABA in the MnPO leading to greater sympathoexcitation during CIH. These hypotheses will be tested using an integrative approach combining whole animal experiments employing virally mediated shRNA knockdown, in vivo sympathetic nerve and single unit recording, in vitro patch clamp recording, and laser capture microdissection with qRT-PCR. Optogenetic approaches will be used to test the role of the SFO-MnPO pathway in CIH hypertension using promoters that are specific for glutamate expressing neurons.