Local effects of substance P on respiratory regulation in the rat medulla oblongata.

The effect of substance P (SP) and the SP antagonist [D-Pro2,D-Trp7,9]-SP on basal ventilation was investigated in halothane-anesthetized rats. Microinjections of SP (0.4-1.5 nmol) into the ventrolateral medulla oblongata (VLM), (nuclei gigantocellularis, facialis, ambiguus, and reticularis lateralis) or into the dorsomedial medulla oblongata (DM, nucleus tractus solitarius) and its ventral surroundings dose dependently increased tidal volume (VT) and/or minute ventilation. In sensitive areas, the ventilatory stimulation was initiated within minutes, peaked around 8-10 min, and slowly returned to normal over 30-45 min after the injection. In the VLM sites, the increase in VT was generally accompanied by a decrease in respiratory frequency (f), whereas in the DM, f increased in parallel with VT. Furthermore, within the VLM, the respiratory response patterns differed with the definite location of the SP injection. A shortening of inspiratory time was observed in the ventromedial part, the ventrolateral portion of the nucleus paragigantocellularis and ventral to the nucleus facialis. In contrast, a lengthening of expiratory time was seen when SP was injected or applied more laterally along the ventral portion of nucleus facialis and near or directly on the ventral medullary surface. Application of [D-Pro2, D-Trp7,9]SP before or after SP completely antagonized the excitatory effects of SP on ventilation. The SP antagonist administered into the VLM decreased the ventilatory response to hypoxic breathing but caused no change during hyperoxic conditions.     

Fluorescence location of RVLM kainate microinjections that alter the control of breathing

Kainic acid (4.7 mM) applied to the rostral ventrolateral medulla (RVLM) surface decreases phrenic output, CO2 sensitivity, and blood pressure in chloralose-urethan-anesthetized, vagotomized, paralyzed, glomectomized, servoventilated cats. In this study using the same preparation, bilateral 50- to 100-nl kainate injections just below the RVLM surface better localized these responses topographically. The physiological responses to unilateral 10-nl kainate injections were then correlated with anatomic location determined by fluorescent microbeads (0.5 micron diam). Many sites were associated with no effect, a few rostral and caudal sites with increased phrenic activity, and cluster of sites with decreased phrenic activity often to apnea, decreased CO2 sensitivity, and decreased responses to carotid sinus nerve stimulation. Blood pressure was unaffected. These sites, within 400 microns of the surface, were ventral to the facial nucleus, ventrolateral to the nucleus paragigantocellularis lateralis, caudal to the superior olive, and rostral to the retrofacial nucleus. They appeared to be within the recently described retrotrapezoid nucleus, which contains cells with respiratory-related activity and projections to the dorsal and ventral respiratory groups. Cells within this site appear able to provide tonic input to respiration and to affect peripheral and central chemoreception.     

Substance P in the ventrolateral medulla oblongata regulates ventilatory responses.

Local injection of substance P (SP) into the ventral portion of the nucleus gigantocellularis, nucleus reticularis lateralis, and nucleus retrofacialis of the ventrolateral medulla oblongata (VLM) or direct application on the ventral surface of the medulla oblongata caused marked stimulation of tidal volume (VT) and/or minute ventilation (VE). The ventilatory response to hypoxia was significantly blunted after SP in the VLM but not in the dorsal medulla oblongata (DM) (nucleus tractus solitarius). The SP antagonist [D-Pro2,D-Trp7,9]SP almost completely inhibited this response when applied locally to a wide area of the superficial layer of the VLM but not of the DM. Unilateral or bilateral application of 0.3-1.5 nmol of the SP antagonist in the VLM (corpus trapezoideum and the caudal region extending from the rootlets of the nucleus hypoglossus to the first cervical segment) markedly attenuated the response to a 5% CO2 inhalation. The inhibition of the CO2 response was seen after [D-Pro2,D-Trp7,9]SP in the rostral areas of the medulla oblongata corresponding to the corpus trapezoideum and the caudal region extending from the rootlets of the nucleus hypoglossus to the first cervical segment of the cervical cord. Electric somatosensory-induced ventilatory stimulation could be depressed by approximately 70% by [D-Pro2,D-Trp7,9]SP locally applied on the surface of the VLM. We conclude that SP is involved in the hypoxic, hypercapnic, and somatosensory ventilatory responses in the rat. However, these respiratory reflexes are mediated via different neuronal pools in the medulla oblongata, mainly the VLM.     

Distribution of NT-IR perikarya in the brain of the guinea pig with special reference to cardiovascular centers in the medulla oblongata

Summary The occurrence and distribution of neurotensin-immunoreactive (NT-IR) perikarya was studied in the central nervous system of the guinea pig using a newly raised antibody (KN 1). Numerous NT-IR perikarya were found in the nuclei amygdaloidei, nuclei septi interventriculare, hypothalamus, nucleus parafascicularis thalami, substantia grisea centralis mesencephali, ventral medulla oblongata, nucleus solitarius and spinal cord. The distribution of NT-IR perikarya was similar to that previously described in the rat and monkey. In the gyrus cinguli, hippocampus and nucleus olfactorius, though, no NT-IR neurons were detected in this investigation. Additional immunoreactive perikarya, however, were observed in areas of the ventral medulla oblongata, namely in the nucleus paragigantocellularis, nucleus retrofacialis and nucleus raphe obscurus.The relevance of the NT-IR perikarya within the ventral medulla oblongata is discussed with respect to other neuropeptides, which are found in this area, and to cardiovascular regulation.Abbreviations abl nucleus amygdaloideus basalis lateralis - abm nucleus amygdaloideus basalis medialis - acc nucleus amygdaloideus centralis - aco nucleus amygdaloideus corticalis - ahp area posterior hypothalami - ala nucleus amygdaloideus lateralis anterior - alp nucleus amygdaloideus lateralis posterior - ame nucleus amygdaloideus medialis - atv area tegmentalis ventralis - bst nucleus proprius striae terminalis - CA commissura anterior - CC corpus callosum - cgld corpus geniculatum laterale dorsale - cglv corpus geniculatum laterale ventrale - cgm corpus geniculatum mediale - CHO chiasma opticum - CI capsula interna - co nucleus commissuralis - cod nucleus cochlearis dorsalis - cp nucleus caudatus/Putamen - cs colliculus superior - cu nucleus cuneatus - dmh nucleus dorsomedialis hypothalami - DP decussatio pyramidum - em eminentia mediana - ent cortex entorhinalis - epi epiphysis - FLM fasciculus longitudinalis medialis - fm nucleus paraventricularis hypothalami pars filiformis - FX fornix - gd gyrus dentatus - gp globus pallidus - gr nucleus gracilis - hl nucleus habenulae lateralis - hm nucleus habenulae medialis - hpe hippocampus - ift nucleus infratrigeminalis - io oliva inferior - ip nucleus interpeduncularis - LM lemniscus medialis - MT tractus mamillo-thalamicus - na nucleus arcuatus - nls nucleus lateralis septi - nms nucleus medialis septi - npca nucleus proprius commissurae anterioris - ns nucleus solitarius - n III nucleus nervi oculomotorii - nt V nucleus tractus spinalis nervi trigemini - ntm nucleus mesencephalicus nervi trigemini - osc organum subcommissurale - P tractus cortico-spinalis - PC pedunculus cerebri - PCI pedunculus cerebellaris inferior - pir cortex piriformis - pol area praeoptica lateralis - pom area praeoptica medialis - prt area praetectalis - pt nucleus parataenialis - pvh nucleus paraventricularis hypothalami - pvt nucleus paraventricularis thalami - r nucleus ruber - re nucleus reuniens - rgi nucleus reticularis gigantocellularis - rl nucleus reticularis lateralis - rm nucleus raphe magnus - ro nucleus raphe obscurus - rp nucleus raphe pallidus - rpc nucleus reticularis parvocellularis - rpgc nucleus reticularis paragigantocellularis - sch nucleus suprachiasmaticus - SM stria medullaris thalami - snc substantia nigra compacta - snl substantia nigra lateralis - snr substantia nigra reticularis - ST stria terminalis - tad nucleus anterior dorsalis thalami - tam nucleus anterior medialis thalami - tav nucleus anterior ventralis thalami - tbl nucleus tuberolateralis - tc nucleus centralis thalami - tl nucleus lateralis thalami - tmd nucleus medialis dorsalis thalami - TO tractus opticus - TOL tractus olfactorium lateralis - tpo nucleus posterior thalami - tr nucleus reticularis thalami - trs nucleus triangularis septi - TS tractus solitarius - TS V tractus spinalis nervi trigemini - tvl nucleus ventrolateralis thalami - vmh nucleus ventromedialis hypothalami - vh ventral horn, Columna anterior - zi zona incerta Supported by the Deutsche Forschungsgesellschaft (DFG) SFB 90, Carvas     

Immunocytochemical localization of GABA containing neurons in the ventrolateral medulla oblongata of the rat

Localization of gamma-aminobutyric acid (GABA) in the ventrolateral medulla oblongata of the rat was studied, using antisera directed against GABA molecule fixed to bovine serum albumin. Within the rostral portion of the ventrolateral medulla, GABA-like immunoreactive neurons were found in the lateral wing of the raphe magnus and in the region of the paragigantocellular reticular nucleus. In the caudal portion of the ventrolateral medulla, a lesser number of GABA-stained neurons were found in the region around the nucleus reticularis lateralis. GABA-like immunoreactive punctate structures were also found throughout the ventrolateral medulla. These results provide further evidence for the existence of GABAergic neurons in the ventrolateral medulla oblongata of the rat.     

心外膜应用腺苷时c—fos在脊髓延髓和丘脑中的表达

在１２只切断两侧缓冲神经和迷走神经的麻醉大鼠,观察了心外膜应用腺苷对脊髓,延髓和丘脑ｃ－ｆｏｓ原部基因表达的影响。结果显示：心外膜应用腺苷组大鼠,动脉血压和心率无明显变化;脊髓Ｔ３节段背角,延髓巨细胞旁外侧核以及丘脑的腹后外侧核,后核,中央外侧核和束旁核等部位Ｆｏｓ蛋白样免疫阳性反应神经元显著增加;而在溶剂对照组大鼠,仅见少数ＦＬＩ细胞。     

辣椒素对大鼠延髓腹外侧头端区神经元电活动的影响

在35只切断两侧缓冲神经的麻醉大鼠,应用细胞外记录的电生理学方法,观察颈总动脉注射辣椒素(capsaicin)对延髓腹外侧头端区(RVLM)巨细胞旁外侧核(PGL)自发电活动的影响。所得结果如下:(1)颈动脉注射辣椒素(10μmol,01ml),MAP由1074±013升至1256±021kPa(P<0001);HR由374±4增至395±5bpm(P<0001);30个PGL神经元自发放电单位的放电频率由126±07增至209±11spikes/s(P<0001)。(2)在10个放电单位,应用辣椒素受体阻断剂钌红(rutheniumred;200mmol,01ml)后,明显抑制辣椒素的上述效应。以上结果提示,辣椒素可能通过激活RVLM神经元上的辣椒素受体,进而兴奋PGL神经元     

肾缺血增强大鼠延髓腹外侧头端区神经元电话动和Fos蛋白表达

在67只切断两侧缓冲神经的麻醉Sprague-Dawley大鼠,应用细胞外记录的电生理方法和免疫组织化学技术,分别观察肾缺血对延髓腹外侧头端区巨细胞旁外侧核神经元自发放电活动和Fos蛋白表达的影响.所得结果如下(1)左肾动脉阻断后,28个单位的放电频率由11.40±1.08增至21.1±1.74spikes/s(P<0.001),血压和心率无明显变化(P>0.05);(2)在17个放电单位中,应用腺苷受体拮抗剂8-苯茶碱(8-phenyltheophylline,10mg/kg)可明显抑制肾缺血的兴奋效应(P<0.05);(3)肾缺血后,延髓腹外侧头端区的Fos蛋白样免疫反应神经元显著增加(P<0.01);(4)预先应用8-苯茶碱可明显减弱肾缺血所激活的Fos蛋白表达反应(P<0.05).以上结果提示肾缺血增强延髓腹外侧头端区神经元的放电活动和Fos蛋白表达,而此作用可能与肾脏缺血所产生的腺苷激活肾内感受器有关.     

Effect of stress exposure on the activation pattern of enkephalin-containing perikarya in the rat ventral medulla

We examined the effects of acute and chronic psychogenic stress on the activation pattern of enkephalin-containing perikarya in the rat ventrolateral medulla. Rats allocated to the chronic stress groups were subjected to 90 min of immobilization for 10 days. On the 11th day, the chronically stressed rats were exposed to homotypic (90-min immobilization) or to heterotypic but still psychogenic (90-min immobilization coupled to air jet stress) stress. The acute stress group was subjected once to an acute 90-min immobilization. For each group, the rats were anesthetized either before stress (time 0) or 90, 180, and 270 min after the onset of stress. Brain sections were then processed using immunocytochemistry (Fos protein) followed by radioactive in situ hybridization histochemistry (enkephalin mRNA). Following immobilization, the acute group displayed a marked increase in the number of activated enkephalin-containing perikarya within the paragigantocellularis and lateral reticular nuclei. This level of activation was sustained up to 180 min following the onset of the immobilization stress and had returned to baseline levels by 270 min from the initiation of the stress. However, this stress-induced activation of enkephalin-containing perikarya of the ventrolateral medulla was not seen following either homotypic or heterotypic stress in the chronically stressed group. These results provide evidence that enkephalin-containing perikarya of the ventrolateral medulla may constitute a potential circuit through which they regulate some aspect of the stress responses. Conversely, this enkephalinergic influence from the ventrolateral medulla was shown to be absent following chronic stress exposure. This would suggest a decrease in enkephalin inhibitory input originating from the ventrolateral medulla, thereby allowing a neuroendocrine and/or autonomic response to chronic stress.     

The effects of ouabain in the medullary site of the hypotensive action of clonidine

Blood pressure was studied in pentobarbital anesthetized rats and cats after central administration of ouabain. Intracerebroventricular (i.c.v.) injections caused a classical biphasic effect, a short lasting hypotension followed by a hypertensive phase. When injected directly into the nucleus reticularis lateralis region (NRL), ouabain (0.01–2 μg/kg) caused a dose-dependent pressor effect. In the same region, kryptofix 221, a sodium complexing agent, produced a fall in blood pressure. Moreover, central administration of ouabain prevented the hypotensive effect of i.v. clonidence whereas the central hypotensive effect of muscimol was not affected. It is concluded that sodium movements play an important role in the blood pressure regulation within the NRL region. We also report here that ouabain antagonizes the hypotensive effect of clonidine suggesting that sodium movement might be the essential link of this action.     

Effects on respiratory pattern of focal cooling in the medulla of the dog

Studies in cats have shown that, in addition to respiratory neuron groups in the dorsomedial (DRG) and ventrolateral (VRG) medulla, neural structures in the most ventral medullary regions are important for the maintenance of respiratory rhythm. The purpose of this study was to determine whether a similar superficially located ventral region was present in the dog and to assess the role of each of the other regions in the canine medulla important in the control of breathing, in 20 anesthetized, vagotomized, and artificially ventilated dogs, a cryoprobe was used to cool selected regions of the medulla to 15-20 degrees C. Respiratory output was determined from phrenic nerve or diaphragm electrical activity. Cooling in or near the nucleus of the solitary tract altered timing and produced little change in the amplitude or rate of rise of inspiratory activity; lengthening of inspiratory time was the most common timing effect observed. Cooling in ventrolateral regions affected the amplitude and rate of rise of respiratory activity. Depression of neural tidal volume and apnea could be produced by unilateral cooling in two ventrolateral regions: 1) near the nucleus ambiguus and nucleus para-ambiguus and 2) just beneath the ventral medullary surface. These findings indicate that in the dog dorsomedial neural structures influence respiratory timing, whereas more ventral structures are important to respiratory drive.     

CCK—8、M—ENK免疫反应结构在猫延髓吻侧腹侧区的分布

用免疫组化 ABC 技术,观察了八肽缩胆囊素(CCK—8),甲硫氨酸脑啡肽(M—ENK)免疫反应(IR)结构在猫延髓吻侧腹侧区的分布。结果表明:CCK—8—IR 细胞分内、外两群:内侧群细胞分布于巨细胞网状核(NGc)、旁巨细胞外侧核(PGL)以及下橄榄核背外侧的网状结构,从吻侧向尾侧逐渐减少;外侧群细胞分布于外侧网状核(LRN)及其背内侧网状结构,从吻侧向尾侧逐渐增多。在中缝苍白核(Rpa)、中缝大核(Rm)仅见少量 CCK—8—IR 细胞。察见 CCK—8—IR纤维主要有3种:粗、细和终末前纤维。CCK—8—IR 纤维在面后核、疑核以及二核紧邻的网状结构最为密集;在 PGL 密度中等;在 NGc、LRN、Rpa 和 Rm 稀疏分布。M—ENK—IR 细胞和纤维分布于 Rpa、Rm、NGc、PGL 和 LRN,此外在面后核、疑核以及二核紧邻的网状结构可见较密集的纤维。     

Tremor following intracerebral carbachol injection. 1.Carbachol sensitivity of various brain structures]

Tremor produced by intracerebral injection of carbachol. I. Susceptibility of different brain areas to carbachol Microinjections of carbachol into the lateral ventricle of rats caused tremor depending on dose. Intensity and duration of motor effects after injection of carbachol (30 mug/3 mul bilateral) into different brain areas were found to depend on localization: strongest tremor was elicited by injections into the nucleus caudatoputamen and cortex cerebri, moderate tremor by administration into the substantia nigra reticularis, globus pallidus and thalamic brain regions. Target areas of mean sensitivity were demonstrated in more rostral and caudal parts of the formatio reticularis. The injection of carbachol into the nucleus ruber, nucleus linearis and substantia nigra compacta brought about lowest tremor values. Ablation of the site of injection from the remaining brain abolished tremor induced by carbachol contrary to the tremor induced by oxotremorine.     

顶核—延髓头端腹外侧区系统参与前庭降压降心率反应

实验用乌拉坦麻醉、箭毒化、人工呼吸的大鼠。将神经元胞体兴奋剂L-谷氨酸钠(Glu)微量注入顶核或前庭上核均引起血压下降;心率减慢。该顶核-和前庭上核-降压降心率反应均可被延髓头端腹外侧区内注射GABA受体阻断剂荷包牡丹碱阻断。顶核内注射普鲁卡因也能阻断Glu兴奋前庭上核的心血管反应。以上结果提示前庭-降压降心率反应可能通过顶核-延髓头端腹外侧区系统实现。静脉注射甲基阿托品也能衰减Glu兴奋顶核的心血管反应,显示迷走神经也参与前庭-顶核降压降心率反应。     

Immunocytochemical localization of GABA containing neurons in the ventrolateral medulla oblongata of the rat

Summary Localization of -aminobutyric acid (GABA) in the ventrolateral medulla oblongata of the rat was studied, using antisera directed against GABA molecule fixed to bovine serum albumin. Within the rostral portion of the ventrolateral medulla, GABA-like immunoreactive neurons were found in the lateral wing of the raphe magnus and in the region of the paragigantocellular reticular nucleus. In the caudal portion of the ventrolateral medulla, a lesser number of GABA-stained neurons were found in the region around the nucleus reticularis lateralis. GABA-like immunoreactive punctate structures were also found throughout the ventrolateral medulla. These results provide further evidence for the existence of GABAergic neurons in the ventrolateral medulla oblongata of the rat.     

Central galanin and N-terminal galanin fragment induce c-Fos immunoreactivity in the medulla oblongata of the anesthetized rat

This immunohistochemical study analyzed the c-Fos expression (c-Fos-ir) induced by galanin injections. Galanin and N-terminal galanin fragment (1-15) induced a significant increase of c-Fos expression (c-ir) within the medulla oblongata 90 min and 6 h. after intracisternal injections. This expression has been studied mainly in the nucleus of the solitary tract and in the ventrolateral medulla showing different temporal profiles for both peptides. The presence of c-Fos-ir in TH-positive cells was analyzed in all the groups. These results may be relevant to understand the role of galanin in several functions including central cardiovascular control.     

The blood pressure effects of alpha-adrenoceptor antagonists injected in the medullary site of action of clonidine: The nucleus reticularis lateralis

We administered a series of alpha-blocking drugs to the nucleus reticularis lateralis (NRL) of the medulla oblongata, the main site for the hypotensive action of clonidine. These experiments were performed on pentobarbital anaesthetized cats. Drugs were injected through a needle which was stereotaxically inserted. Prazosin (6 nmol) was hypertensive (MBP=+25±8%), corynanthine had no effect and AR-C239 (7 nmol), another alpha₁- blocker, was hypotensive (MBP=−16±3.5%).The alpha₂-blockers, yohimbine and idaxozan, were hypotensive. The blood pressure effects of alpha-blocking drugs directly microinjected in the nucleus reticularis lateralis cannot be simply related to their selectivity for a particular subtype of alpha-receptors.     

Vasoactive intestinal polypeptide immunoreactivity in the spinal cord of the guinea pig

Summary The distribution of vasoactive intestinal polypeptide-immunoreactive (VIP-IR) neurons in the lower medulla oblongata and the spinal cord has been analyzed in guinea pigs. This study includes results obtained by colchicine treatment and transection experiments. In the spinal cord, numerous VIP-IR varicosities were observed in the substantia gelatinosa of the columna dorsalis; some were also found in the substantia intermedia and the columna anterior. The spinal VIP-IR nerve fibers were mainly of intraspinal origin and oriented segmentally. VIP-IR nuclei in the spinal cord extended dorsally into corresponding regions of the caudal medulla oblongata, namely from the substantia intermedia medialis and lateralis into the vagus-solitarius complex and from the nucleus spinalis lateralis into the area of the nucleus reticularis lateralis. Additional VIP-IR perikarya were observed in the pars caudalis of the nucleus spinalis nervi trigemini. The VIP-IR nuclei within the caudal medulla oblongata probably form a continuous system with those localized within the spinal cord. They may be involved functionally in the modulation of cardiovascular and respiratory regulation in the guinea pig.Supported by the DFG, Carvas SFB 90     

Role of pressor mechanisms from the NTS and CVLM in control of arterial pressure

In the present study, we investigated the effects of inhibition of the caudal ventrolateral medulla (CVLM) with the GABA(A) agonist muscimol combined with the blockade of glutamatergic mechanism in the nucleus of the solitary tract (NTS) with kynurenic acid (kyn) on mean arterial pressure (MAP), heart rate (HR), and regional vascular resistances. In male Holtzman rats anesthetized intravenously with urethane/chloralose, bilateral injections of muscimol (120 pmol) into the CVLM or bilateral injections of kyn (2.7 nmol) into the NTS alone increased MAP to 186 +/- 11 and to 142 +/- 6 mmHg, respectively, vs. control: 105 +/- 4 mmHg; HR to 407 +/- 15 and to 412 +/- 18 beats per minute (bpm), respectively, vs. control: 352 +/- 12 bpm; and renal, mesenteric and hindquarter vascular resistances. However, in rats with the CVLM bilaterally blocked by muscimol, additional injections of kyn into the NTS reduced MAP to 88 +/- 5 mmHg and mesenteric and hindquarter vascular resistances below control baseline levels. Moreover, in rats with the glutamatergic mechanisms of the NTS blocked by bilateral injections of kyn, additional injections of muscimol into the CVLM also reduced MAP to 92 +/- 2 mmHg and mesenteric and hindquarter vascular resistances below control baseline levels. Simultaneous blockade of NTS and CVLM did not modify the increase in HR but also abolished the increase in renal vascular resistance produced by each treatment alone. The results suggest that important pressor mechanisms arise from the NTS and CVLM to control vascular resistance and arterial pressure under the conditions of the present study.