中枢酸敏感离子通道对呼吸的调节及机制研究

发布时间：2018-01-12 07:27

本文关键词：中枢酸敏感离子通道对呼吸的调节及机制研究　出处：《复旦大学》2010年博士论文　论文类型：学位论文

【摘要】：目的：呼吸运动是一种受呼吸中枢调节的节律性的活动。中枢化学感受器(central chemoreceptors)感受细胞外pH和CO2的浓度变化,将化学信号转化电信号,传导给呼吸中枢的运动神经元,完成对呼吸运动的调节。中枢化学感受器主要存在于延髓腹外侧区(ventrolateral medulla, VLM),近几年下丘脑外侧区(lateral hypothalamus, LH)的Orexin神经元在中枢化学感受中的作用也逐渐引起大家的关注。目前,中枢化学感受器感受细胞外H+浓度变化的离子通道机制还不明确,也一直是呼吸生理学研究的重点。1997年,Waldmann,R.克隆了第一个酸敏感离子通道(acid-sensing ion channels, ASICs),这是一类H+门控的阳离子通道。研究发现,ASICs参与机体许多生理和病理生理过程,如：学习记忆,痛觉、视觉、听觉、味觉的感受,脑缺血损伤等,但是ASICs在呼吸的中枢调节中的作用却知之甚少。本课题假设：1ASICs在VLM和LH都有表达并参与对呼吸的调节；2.LH的Orexin神经元通过其胞膜上的ASICs通道感受细胞外pH值的变化,并调节呼吸运动。中枢的化学感受调节是快速调节维持机体酸碱平衡、内环境稳定的关键环节。中枢性呼吸衰竭、中枢性呼吸节律紊乱、先天性中枢性肺换气不足综合征等呼吸中枢调节紊乱的疾病对机体造成的危害是致命的,在一些肺和呼吸道病变所引发的疾病如慢性阻塞性肺疾病、阻塞性睡眠呼吸暂停低通气综合征等疾病的病理过程中也伴有呼吸中枢调节功能的异常。因此,探讨呼吸中枢调控机制不仅是呼吸生理学的理论突破,对呼吸系统疾病的病理过程研究和临床预防与治疗也具有重要的意义。方法：1.实验用新生(1～5d)和成年(6～8w)SD大鼠,雌雄不计,取延髓部位,用免疫组织化学ABC方法、免疫荧光双标、confoca1、westernblot和图像分析技术,观察和比较ASIC1和ASIC2a亚单位在大鼠延髓和下丘脑的表达,Orexin A神经元在下丘脑的表达,Orexin A神经纤维在延髓的投射,Orexinl型受体(orexin type1receptor, OX1R)在延髓的表达以及ASIC1,ASIC2a亚单位与神经纤维丝-H的共表达、ASIC1与ASIC2a亚单位的共表达、ASIC1与Orexin的共表达。2.成年(6～8w)SD大鼠腹腔麻醉后,应用电生理实验,采用脑立体定位和中枢核团微量注射技术,首先观察在延髓腹外侧区(VLM)和下丘脑外侧区(LH)微量注射不同pH值的人工脑脊液对膈神经放电活动、血压和心率的影响。然后在LH微量注射ASICs的非特异性阻断剂Amiloride或ASICla选择性阻断剂PcTX1后,观察酸化LH对呼吸和心血管系统的调节作用。观察在侧脑室微量注射Orexin A对膈神经放电活动、血压和心率的影响。3.成年(6～8w)SD大鼠腹腔麻醉后,应用电生理实验,采用脑立体定位和中枢核团微量注射技术,在LH注射酸化人工脑脊液激活ASICs,同时在延髓的孤束核微量注射SB408124阻断OX1R后,观察膈神经放电活动、血压、心率的变化。4.采用脑立体定位和中枢核团微量注射技术,成年(6-8w)SD大鼠的LH区微量注射Orexin-SAP特异性损毁该核团的Orexin能神经元,观察大鼠摄食和体重的变化；应用电生理实验观察核团损毁后,动物的基础呼吸频率、血压和心率的变化,以及酸化刺激LH区对心血管呼吸系统的调节。结果 1. ASIC1和ASIC2a在成年和新生SD大鼠延髓的表达：(1)免疫组化的结果显示：ASIC1和ASIC2a免疫反应阳性细胞在成年鼠的延髓有广泛分布：ASIC1和ASIC2a免疫反应阳性细胞在第四脑室旁的中间核(intercalated nuclus of medulla, In)以及延髓背侧的楔束副核(external cuneate nuclus, ECu)都有散在分布,在延髓的头端腹外侧区则有着广泛分布。ASIC1和ASIC2a免疫反应阳性细胞在新生鼠延髓的表达略有不同：ASIC1免疫反应阳性细胞主要分布在VLM区,在延髓背侧的ECu也有散在表达；ASIC2a免疫反应阳性细胞也主要分布在VLM区,但是在延髓背侧未观察到ASIC2a免疫反应阳性细胞。在VLM区,成年鼠组的ASIC2a免疫反应的相对光密度(relative optical density, ROD)和阳性细胞数值均低于新生鼠组的(p0.001,n=6)；成年鼠组的ASIC1免疫反应的ROD值高于新生鼠组(p0.001,n=6)而细胞计数统计结果显示成年鼠组的ASIC1免疫反应阳性细胞数少于新生鼠组(p0.001,n=6)。(2)应用Western blot的方法：在成年鼠和新生鼠延髓都检测到ASIC1和ASIC2a蛋白的表达,ASIC1和ASIC2a在成年鼠组延髓的蛋白含量均低于新生鼠组(p0.01,p0.001,n=6)。(3)应用免疫荧光双染和免疫荧光共聚焦实验方法,在VLM区观察到了ASIC1和ASIC2a与神经纤维丝-H的共表达,以及ASIC1和ASIC2a的共表达。 2. ASIC1和ASIC2a在成年SD大鼠下丘脑的表达：免疫组织化学实验结果显示：ASIC1和ASIC2a两个亚单位在成年SD大鼠的下丘脑广泛表达：ASIC1和ASIC2a免疫反应阳性细胞在下丘脑背侧区(dorsal hypothalamus area, DA)和下丘脑外侧区(LH)都有分布,但是两者的分布存在差异性。细胞计数结果显示：LH区的ASIC1免疫反应阳性细胞数多于DA区(p0.001,n=7),ASIC2a免疫反应阳性细胞数在DA区多于在LH的分布(p0.001,n=7)；在DA区ASIC1免疫反应阳性细胞数少于ASIC2a (p0.01,n=7),LH区ASIC1免疫反应阳性细胞数多于ASIC2a(p0.001,n=7)。对ASIC1和ASIC2a免疫反应阳性细胞的相对光密度(ROD)值的分析未发现两者在DA区和LH区存在明显差异。 3.在VLM区和LH区微量注射酸化人工脑脊液对呼吸及心血管系统的调节作用：(1)在SD大鼠的单侧VLM区注射不同酸化程度的人工脑脊液(0.1μ1),pH值分别为pH7.4、6.5、6.0、5.5、5.0、4.5,与pH7.4组相比,观察到pH6.5和pH6.0的人工脑脊液可以增加膈神经放电强度(p0.05,n=12),对呼吸频率、血压、心率均没有明显的影响。(2)在SD大鼠的单侧LH区注射不同酸化程度的人工脑脊液(0.1μ1),pH值分别为pH7.4、6.5、5.5、4.5,与pH7.4组相比,观察到pH6.5的人工脑脊液可以明显增大膈神经放电强度(p0.001,n=6),对平均动脉压、心率和呼吸频率没有明显影响。(3)在LH区微量注射ASICs非特异性阻断剂Amiloride (10mM,0.1μl)或ASIC1a特异性阻断剂PcTX1(10nM,0.1μl)之后立刻注射pH6.5人工脑脊液(0.1μ1),明显阻断了pH6.5人工脑脊液的增大膈神经的效应(p0.001,n=6),而单独注射Amiloride或PcTX1对呼吸没有明显抑制作用。各个用药组对呼吸频率、血压、心率没有明显的影响。 4. Orexin A对呼吸和心血管系统的调节作用：(1)免疫组化结果显示：Orexin A免疫反应阳性纤维存在于延髓的孤束核(nucleus tractus solitary, NTS),并且在NTS也观察到OX1R免疫反应阳性神经元的存在。(2)单侧侧脑室微量注射不同浓度的Orexin A(10、100、10001μg/ml,5μl),剂量依赖性地增强膈神经放电强度、升高动脉血压、加快心率,但对呼吸频率没有影响。三个浓度组的膈神经放电强度分别与生理盐水对照组相比都有统计意义(p0.05,p0.01,p0.001,n=6)。100、1000μg/ml两个浓度组的平均动脉压与生理盐水对照组相比有统计意义(p0.05,p0.01,n=6)。100、1000μg/ml浓度组的心率与生理盐水对照组相比有统计意义(p0.01,p0.01,n=6)。 5. ASIC1与Orexin A在LH区的共表达,以及它们在呼吸调节中的相互作用：(1)应用免疫组化实验在LH区观察到Orexin A免疫反应阳性细胞。应用免疫荧光双染和免疫荧光共聚焦实验方法,在LH区观察到ASIC1亚单位与Orexin A的共表达。(2)在单侧LH区注射pH6.5人工脑脊液(0.1μl),同时在同侧NTS注射SB408124(0.1μl)与注射pH7.4的人工脑脊液(0.1μl)相比,明显阻断了pH6.5人工脑脊液的增加呼吸强度的效应(p0.01,n=6),而单侧LH区注射pH7.4的人工脑脊液,同时同侧NTS注射SB408124对呼吸强度没有影响。(3)双侧LH区注射Orexin-SAP(0.43mg/ml,400nl/侧)两周后,观察到损毁组大鼠的体重明显下降,应用尼氏染色和免疫组化方法,观察到两周后LH区的Orexin A神经元数量明显减少,仅有少量残存的Orexin A神经元。应用电生理的实验技术,观察到损毁组大鼠与Blank-SAP对照组相比平均动脉压下降,心率减慢(p0.01,n=6),呼吸频率没有变化。在损毁组大鼠的单侧LH区注射pH6.5人工脑脊液,对膈神经放电强度不再有兴奋作用。结论： 1.AISC1和ASIC2a亚单位在SD大鼠的延髓呼吸相关核团VLM区和下丘脑的LH区都有表达。随着发育成熟,ASIC1和ASIC2a亚单位蛋白表达量在成年鼠延髓的比在新生鼠减少,虽然ASIC1细胞数量减少,但是ASIC1免疫反应阳性细胞的相对光密度值(ROD)增加,提示成年鼠延髓单个细胞上的ASIC1表达增加。ASIC2a的ROD和细胞数量在成年鼠VLM区都比在新生鼠减少。ASICs亚单位表达量的变化可能与不同发育阶段中枢对化学刺激的敏感性不同有关。ASIC1和ASIC2a在LH区都有表达,但是以ASIC1亚单位为主。提示下丘脑的LH区可能是新发现的中枢化学敏感区。实验结果为探讨ASICs在呼吸中枢调节中的作用提供了形态学基础。 2.在延髓VLM区和下丘脑LH区适度的酸化刺激都可以增加呼吸强度。观察到酸化LH区引起的呼吸兴奋效应更明显,其有效刺激的酸化程度为pH6.5,与ASIC la的pHo.5相符。ASICs的非特异性阻断剂Amiloride和ASIC1a的特异性阻断剂PcTX1几乎都可以完全阻断酸化LH区引起的兴奋呼吸的效应。在生理pH值范围内,单独阻断LH区的ASICs通道对呼吸并没有抑制作用。提示ASICs不仅在LH区存在,并且感受局部的化学变化,然后调节呼吸活动,在此调节过程中ASIC1a亚单位起主要作用。 3. ASIC1在下丘脑LH区的Orexin能神经元上表达。LH区的Orexin(?)神经元损毁后,酸化刺激LH区不再有兴奋呼吸的作用。提示LH区的Orexin(?)神经元可以感受局部的化学变化,可能是一类新发现的中枢化学感受器。而ASICs在下丘脑对呼吸的调节作用正是通过Orexin(?)神经元实现的。侧脑室微量注射外源性的OrexinA可以增加呼吸强度,Orexin能神经纤维和OX1R在延髓NTS有表达。提示Orexin有可能通过向NTS的投射发挥对呼吸的调节作用。在NTS阻断OX1R可以抑制酸化LH区增加呼吸强度的效应,而单独阻断NTS的OX1R对呼吸活动没有影响。说明酸化刺激LH区,同时阻断NTS的OX1R所产生的效应并非两种相反作用的相互抵消,进一步说明酸化LH区是通过兴奋Orexin神经元,释放Orexin与延髓NTS的OX1R结合,发挥兴奋呼吸的效应。
[Abstract]:Objective: breathing is a central respiratory regulation of rhythmic activity. Central chemoreceptor (central chemoreceptors) feel the concentration changes of extracellular pH and CO2, the chemical signal into electrical signal, transmitted to the motor neurons of the respiratory center, complete adjustment of the respiratory motion. Central chemoreceptor mainly exists in the ventrolateral medulla (ventrolateral medulla, VLM), in recent years, the lateral hypothalamic area (lateral hypothalamus, LH) of Orexin neurons in central chemoreception in gradually attracted attention. At present, the central chemoreceptor ion channel mechanism of extracellular H+ concentration of the feeling is still not clear, has been the research respiratory physiology focuses on.1997, Waldmann, R. cloning of the first acid sensing ion channels (acid-sensing ion, channels, ASICs), which is a kind of H+ gated cation channel research. Found that ASICs participates in many physiological and pathological processes, such as learning and memory, pain, vision, hearing, taste feeling, cerebral ischemia injury, but the role of ASICs in regulation of respiration in the CNS is poorly understood. This research hypothesis: the expression of 1ASICs and involved in the regulation of respiration in VLM and LH; 2.LH Orexin neurons through ASICs channels, its cell membrane feel the change in extracellular pH and regulate respiratory movement. Central chemoreceptor regulation is rapidly adjust and maintain the acid-base balance of the body, the key link in stable environment. Central respiratory failure, central respiratory rhythm disorders, congenital central pulmonary damage hypoventilation syndrome and other diseases of the respiratory central regulation disorders can lead to fatal, in
Some caused lung and respiratory diseases such as chronic obstructive pulmonary disease, obstructive sleep apnea hypopnea syndrome and other pathological process of disease also accompanied by abnormal respiratory function. Therefore, to explore the central respiratory regulation mechanism is not only the theoretical breakthrough of respiratory physiology, also has an important significance to the study of pathology the process of respiratory system diseases and clinical prevention and treatment.
Methods: 1. new experiments (1 ~ 5d) and adult (6 ~ 8W) SD rats, regardless of male and female, and medullary parts, using immunohistochemical ABC method, immunofluorescence, confoca1, Westernblot and image analysis technique to observe the expression and comparison of ASIC1 and ASIC2a subunits in the rat hypothalamus and spinal cord. The Orexin A expression in the hypothalamus neurons, Orexin A nerve fibers in the medulla of the projection type Orexinl receptor (orexin, type1receptor, OX1R) in the medulla and expression of ASIC1, co expression of ASIC2a subunit of neurofilament -H, co expression of ASIC1 and ASIC2a subunit,.2. adult co expression of ASIC1 and Orexin (6 ~ 8W) SD rats after intraperitoneal anesthesia, application of electrophysiological experiments, using stereotaxic and nucleus microinjection technique, first observed in the ventrolateral medulla (VLM) and lateral hypothalamic area (LH) injection of artificial cerebrospinal fluid with different pH values on the diaphragm of God The discharge activity, blood pressure and heart rate. Then in the blocking of non-specific LH microinjection of ASICs inhibitor Amiloride or ASICla selective antagonist PcTX1, observe the acidification LH regulating effect on respiratory and cardiovascular systems. In the observation of intracerebroventricular injection of Orexin A on the discharges of phrenic nerve activity, effects of.3. on blood pressure and heart rate (adult 6 ~ 8W) SD rats after intraperitoneal anesthesia, application of electrophysiological experiments, using stereotaxic and nucleus microinjection technique, the activation of ASICs in LH injection acidizing artificial cerebrospinal fluid, while blocking OX1R in NTS microinjection of SB408124 medulla after phrenic nerve discharge activity, blood pressure, heart rate changes by.4. stereotaxic and nucleus microinjection technique (6-8W), adult SD rats LH microinjection of Orexin-SAP specific damage to the nuclei of Orexin neurons, and observe the changes of food intake and body weight of rats; Electrophysiological experiments were carried out to observe the changes of basal respiratory rate, blood pressure and heart rate, as well as the regulation of acidification stimulation on the cardiovascular system in LH area after nuclear damage.
Result
1. the expression of ASIC1 and ASIC2a in the medulla of rats in adult and newborn SD: (1) the immunohistochemical results showed that medullary ASIC1 and ASIC2a immunoreactive cells in adult rats are widely distributed: intermediate nuclear ASIC1 and ASIC2a immunoreactive cells in the fourth ventricle (intercalated nuclus of medulla, In) wedge beam and accessory nucleus of the dorsal medulla oblongata (external cuneate, nuclus, ECu) are scattered in the end of the ventrolateral of medulla are widely distributed and.ASIC1 expression of ASIC2a immunoreactive cells in neonatal rat medulla is slightly different: ASIC1 immunoreactive cells were mainly distributed in the VLM region in the back rostral ECu also has scattered expression; ASIC2a immunoreactive cells were mainly distributed in the VLM region, but was not observed in the dorsal medulla of ASIC2a immunoreactive cells in the VLM area, ASIC2a group of immune response in adult rats The relative optical density (relative optical, density, ROD) and positive cells were lower than the value of neonatal rats group (p0.001, n=6); ASIC1 immune response of adult rats ROD group is higher than the value of neonatal group (p0.001, n=6) and the statistical results of cell count shows the number of ASIC1 immunoreactive cells in adult rat group than in neonatal rats group (p0.001, n=6). (2) methods Western blot: in the adult rat and neonatal rat medulla were detected the expression of ASIC1 and ASIC2a protein, ASIC1 and ASIC2a in protein content in adult rat medulla were lower than that of neonatal group (P0.01, p0.001, n=6). (3) immunofluorescence double immunofluorescence staining and confocal experiment method, the co expression of ASIC1 and ASIC2a and neurofilament -H in VLM region was observed, and the co expression of ASIC1 and ASIC2a.
2. the expression of ASIC1 and ASIC2a in the hypothalamus of adult SD rats: immunohistochemical results showed that ASIC1 and ASIC2a two subunits are widely expressed in adult SD rat hypothalamus: ASIC1 and ASIC2a immunoreactive cells in the dorsal hypothalamic area (dorsal hypothalamus, area, DA) and lateral hypothalamic area (LH) are have distribution, but the distribution of differences. Cell counting results showed that the LH area is more than the number of ASIC1 immunoreactive cells (p0.001, n=7) DA, ASIC2a immunoreactive cell number in the DA region than in the distribution of LH (p0.001, n=7); in the DA area is less than the number of ASIC1 immunoreactive cells ASIC2a (P0.01, n=7), LH ASIC1 immunoreactive positive cells than ASIC2a (p0.001, n=7). The relative optical density of ASIC1 and ASIC2a immunoreactive cells (ROD) value analysis showed no obvious difference in both DA and LH regions ISO.
3. in the VLM area and LH area of microinjection of acidification artificial cerebrospinal fluid regulating effect on respiratory and cardiovascular system: (1) in the VLM area of different degree of acidification of unilateral injection of rat SD artificial cerebrospinal fluid (0.1 1), pH = pH7.4,6.5,6.0,5.5,5.0,4.5, compared with group pH7.4, pH6.5 and pH6.0 were observed in artificial cerebrospinal fluid can increase the strength of the phrenic nerve discharge (P0.05, n=12), blood pressure, heart rate of respiratory frequency, there is no effect. (2) in the LH area of different degree of acidification of unilateral injection of rat SD artificial cerebrospinal fluid (0.1 1), pH = pH7.4,6.5,5.5,4.5, compared with pH7.4 group, observed pH6.5 artificial cerebrospinal fluid can increase the strength of the phrenic nerve discharge (p0.001, n=6), the mean arterial pressure, heart rate and respiratory rate had no obvious effect. (3) in the LH region of nonspecific microinjection of ASICs inhibitor Amiloride (10mM, 0.1 L) or ASIC1a inhibitor PcT X1 (10nM, 0.1 L) immediately after pH6.5 injection of artificial cerebrospinal fluid (0.1 ~ 1), significantly blocked pH6.5 artificial cerebrospinal fluid increases phrenic nerve effect (p0.001, n=6), and a single injection of Amiloride or PcTX1 had no obvious inhibitory effect on respiration. Each drug group of respiratory frequency, blood pressure, heart rate is not obvious effect.
4. Orexin A on the regulation of respiratory and cardiovascular system: (1) immunohistochemistry results showed that NTS A Orexin immunoreactive fibers were found in the medulla oblongata (nucleus tractus, solitary, NTS), and in NTS were also observed in OX1R immunoreactive neurons. (2) unilateral intracerebroventricular injection different concentrations of Orexin A (1010010001 g/ml, 5 L), dose dependently enhanced phrenic nerve discharge intensity increased, arterial blood pressure, heart rate and respiratory rate. But there is no effect on phrenic nerve discharge intensity of three concentration group respectively with saline control group has statistical significance compared (P0.05, P0.01 p0.001, n=6,.1001000) g/ml two concentration group the mean arterial pressure and saline control group had statistical significance (P0.05, P0.01, n=6 compared with saline.1001000) heart rate g/ml concentrations were statistically significant compared with the control group (P0.01, P0.01, n =6).
Co expression of 5. ASIC1 and Orexin A in the LH region, and their interactions in the regulation of respiration: (1) immunohistochemical experiment showed that A immunoreactive positive cells in the Orexin area of LH. Using immunofluorescence double staining and confocal immunofluorescence experiments, in the zone of LH co expression of ASIC1 the unit and the Orexin A. (2) in the LH region of pH6.5 injection unilateral artificial cerebrospinal fluid (0.1 L), at the same time on the same side NTS injection of SB408124 (0.1 L) and pH7.4 injection of artificial cerebrospinal fluid (0.1 L), significantly blocked the effect of increasing the respiratory intensity of pH6.5 artificial cerebrospinal fluid (P0.01, n=6), and unilateral LH injection in pH7.4 artificial cerebrospinal fluid, and ipsilateral NTS SB408124 injection had no effect on respiration. (3) bilateral LH injection of Orexin-SAP (0.43mg/ml, 400nl/ side) after two weeks, observed damage rats body weight decreased significantly, using Nissl staining and immunohistochemical method, the concept of Orexin observed that the number of A neurons in area LH decreased obviously after two weeks, only a small amount of residual Orexin A neurons. The application of experimental techniques of electrophysiology, observed damaged rats compared with Blank-SAP control group mean arterial pressure, heart rate (P0.01, n=6), respiratory rate did not change. In the lesioned group in the LH region of pH6.5 injection unilateral artificial cerebrospinal fluid, no longer have excitatory effects on phrenic nerve discharge intensity.
Conclusion:
The 1.AISC1 and ASIC2a subunits in SD rat medullary respiratory related nucleus VLM and LH in the hypothalamus region are expressed. With the maturation of ASIC1 and ASIC2a subunit protein expression in neonatal rats than in adult rat medulla decreased, while the number of ASIC1 cells decreased, but the relative optical density of ASIC1 immunoreactive cells the value (ROD) increased, suggesting that adult rat medullary mononuclear cells on the expression of ASIC1 increased the number of.ASIC2a and ROD cells in the adult rat VLM area than the reduction of.ASICs subunit expression may be related to different developmental stages of the central chemical stimuli sensitivity of different.ASIC1 and ASIC2a were expressed in the LH District in the newborn rat, but with ASIC1 subunit. The hypothalamic LH region may be found in central chemosensitive areas. The experimental results provide a morphological basis for the role of ASICs in the regulation of the respiratory center.
2. in the medulla VLM and hypothalamic LH area moderate acidification can increase the respiration intensity of stimulation. The observed respiratory stimulant effect caused by acidification LH region is more obvious, the degree of acidification of effective stimulus for pH6.5, ASIC and La pHo.5 are nonspecific.ASICs specific blocking agent Amiloride and ASIC1a the effect of blocking agent PcTX1 almost can be completely blocked by acidification induced by LH region excited breathing. In the range of physiological pH, single ASICs channel blocking LH area and had no inhibitory effect on respiration. It suggests that ASICs has not only in the LH region, and feel the local chemical changes, and then adjust the respiratory activity, during the adjusting process of ASIC1a subunit plays a major role.
3. ASIC1 in the hypothalamic LH area Orexin expression of neurons in.LH Orexin (?) neurons damage after acidification stimulated LH area no longer have excited breathing effect. LH region of Orexin (?) neurons can feel local chemical changes, may be a newly discovered class of central chemoreceptor and regulation. The effects of ASICs on respiration in the hypothalamus is through Orexin (?) neurons to achieve. Intracerebroventricular injection of exogenous OrexinA can increase the respiration intensity, Orexin nerve fibers and OX1R in medullary NTS

【学位授予单位】：复旦大学
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R332

【参考文献】