Ruyle Brian C, Lima-Silveira Ludmila, Martinez Diana, Cummings Kevin J, Heesch Cheryl M, Kline David D, Hasser Eileen M
Department of Biomedical Sciences, University of Missouri, Columbia, MO, USA.
Dalton Cardiovascular Research Center, University of Missouri, Columbia, MO, USA.
J Physiol. 2023 Oct;601(19):4309-4336. doi: 10.1113/JP284907. Epub 2023 Aug 26.
The hypothalamic paraventricular nucleus (PVN) is essential to peripheral chemoreflex neurocircuitry, but the specific efferent pathways utilized are not well defined. The PVN sends dense projections to the nucleus tractus solitarii (nTS), which exhibits neuronal activation following a hypoxic challenge. We hypothesized that nTS-projecting PVN (PVN-nTS) neurons contribute to hypoxia-induced nTS neuronal activation and cardiorespiratory responses. To selectively target PVN-nTS neurons, rats underwent bilateral nTS nanoinjection of retrogradely transported adeno-associated virus (AAV) driving Cre recombinase expression. We then nanoinjected into PVN AAVs driving Cre-dependent expression of Gq or Gi designer receptors exclusively activated by designer drugs (DREADDs) to test the degree that selective activation or inhibition, respectively, of the PVN-nTS pathway affects the hypoxic ventilatory response (HVR) of conscious rats. We used immunohistochemistry for Fos and extracellular recordings to examine how DREADD activation influences PVN-nTS neuronal activation by hypoxia. Pathway activation enhanced the HVR at moderate hypoxic intensities and increased PVN and nTS Fos immunoreactivity in normoxia and hypoxia. In contrast, PVN-nTS inhibition reduced both the HVR and PVN and nTS neuronal activation following hypoxia. To further confirm selective pathway effects on central cardiorespiratory output, rats underwent hypoxia before and after bilateral nTS nanoinjections of C21 to activate or inhibit PVN-nTS terminals. PVN terminal activation within the nTS enhanced tachycardic, sympathetic and phrenic (PhrNA) nerve activity responses to hypoxia whereas inhibition attenuated hypoxia-induced increases in nTS neuronal action potential discharge and PhrNA. The results demonstrate the PVN-nTS pathway enhances nTS neuronal activation and is necessary for full cardiorespiratory responses to hypoxia. KEY POINTS: The hypothalamic paraventricular nucleus (PVN) contributes to peripheral chemoreflex cardiorespiratory responses, but specific PVN efferent pathways are not known. The nucleus tractus solitarii (nTS) is the first integration site of the peripheral chemoreflex, and the nTS receives dense projections from the PVN. Selective GqDREADD activation of the PVN-nTS pathway was shown to enhance ventilatory responses to hypoxia and activation (Fos immunoreactivity (IR)) of nTS neurons in conscious rats, augmenting the sympathetic and phrenic nerve activity (SSNA and PhrNA) responses to hypoxia in anaesthetized rats. Selective GiDREADD inhibition of PVN-nTS neurons attenuates ventilatory responses, nTS neuronal Fos-IR, action potential discharge and PhrNA responses to hypoxia. These results demonstrate that a projection from the PVN to the nTS is critical for full chemoreflex responses to hypoxia.
下丘脑室旁核(PVN)对于外周化学反射神经回路至关重要,但所利用的具体传出通路尚不清楚。PVN向孤束核(nTS)发出密集投射,在缺氧刺激后nTS会出现神经元激活。我们推测,投射到nTS的PVN(PVN - nTS)神经元促成了缺氧诱导的nTS神经元激活和心肺反应。为了选择性地靶向PVN - nTS神经元,对大鼠双侧nTS进行纳米注射逆行转运的驱动Cre重组酶表达的腺相关病毒(AAV)。然后我们向PVN纳米注射驱动由设计药物(DREADDs)特异性激活的Gq或Gi设计受体的Cre依赖性表达的AAV,以分别测试PVN - nTS通路的选择性激活或抑制对清醒大鼠缺氧通气反应(HVR)的影响程度。我们使用Fos免疫组织化学和细胞外记录来研究DREADD激活如何影响缺氧引起的PVN - nTS神经元激活。通路激活在中度缺氧强度下增强了HVR,并在常氧和缺氧状态下增加了PVN和nTS的Fos免疫反应性。相比之下,PVN - nTS抑制降低了缺氧后的HVR以及PVN和nTS神经元激活。为了进一步证实对中枢心肺输出的选择性通路效应,在双侧nTS纳米注射C21以激活或抑制PVN - nTS终末之前和之后对大鼠进行缺氧处理。nTS内的PVN终末激活增强了对缺氧的心动过速、交感神经和膈神经(PhrNA)神经活动反应,而抑制则减弱了缺氧诱导的nTS神经元动作电位发放和PhrNA的增加。结果表明PVN - nTS通路增强了nTS神经元激活,并且对于对缺氧的完整心肺反应是必需的。要点:下丘脑室旁核(PVN)促成外周化学反射心肺反应,但特定的PVN传出通路尚不清楚。孤束核(nTS)是外周化学反射的第一个整合位点,并且nTS接受来自PVN的密集投射。PVN - nTS通路的选择性GqDREADD激活被证明可增强清醒大鼠对缺氧的通气反应以及nTS神经元的激活(Fos免疫反应性(IR)),增强麻醉大鼠对缺氧的交感神经和膈神经活动(SSNA和PhrNA)反应。PVN - nTS神经元的选择性GiDREADD抑制减弱了通气反应、nTS神经元Fos - IR、动作电位发放以及对缺氧的PhrNA反应。这些结果表明从PVN到nTS的投射对于对缺氧的完整化学反射反应至关重要。