大鼠延髓腹外侧表面化学感受区与延髓内部核团的联系

本文采用辣根过氧化物酶（ＨＲＰ）组化方法,对大鼠延髓腹外侧表面化学感受区与延髓内部核团之间的神经结构联系进行了系统的探查。实验在３０只麻醉且自主呼吸的雄性ＳＤ大鼠上分四组进行。用ＨＲＰ滤纸分别局部敷贴于延髓腹外侧表面的头端化学感受区（Ｒ区,ｎ＝１０）、尾端化学感受区（Ｃ区,ｎ＝１０）、中间区（Ｉ区,ｎ＝６）和对照区（ｎ＝４）。动物存活２４ｈ后,检查ＨＲＰ标记细胞所在的核团部位。（１）于Ｒ区表面敷贴     

大鼠延髓外周橄榄腹外侧核在中枢化学感受中的作用

本工作采用微量注射、电损毁、电刺激和微电泳的方法,探讨了大鼠延髓外周橄榄腹外侧核（ＬＶＰＯ）的否真正参与中枢化学感受功能。实验在３８只雄性ＳＤ大鼠上进行。结果表明：（１）微量注射酸化人工脑脊液于ＬＶＰＯ可引起隔神经放电活动明显加强。（２）微电泳给予Ｈ＾＋对ＬＶＰＯ的自发放电单位主要引起兴奋反应,微电泳给予Ｈ＾＋引起兴奋反应的部分单位也可被腹外侧表面微量注射酸化人工ＣＳＦ所兴奋。（３）损毁ＬＶＰＯ后     

尾端腹外侧延髓心血管区的功能和调节机制

延髓尾端有两个调节心血管活动的区域,尾端腹外侧延髓除了作为外周压力感受器传入冲动的中继站外,可能还有一压力感受器非依赖的,对头端腹外侧延髓的抑制作用。此外,尾端腹外侧延髓还提供一种非兴奋性氨基酸介导的兴奋冲动,影响头端腹外侧延髓,尾端加压区神经元是支持头端腹外侧延髓交感前运动神经元静息活性的主要突触来源,对维持血管张力起一定作用。     

大鼠延髓腹外侧部微量注射心房钠尿肽的心血管效应

在麻醉大鼠观察了延髓腹外侧部的谷氨酸敏感区(GSA)应用ANP对血压和心率的影响。在GSA应用α-hANP,血压和心率明显降低。低浓度的APⅢ(10~(-6)mol/L)仅引起血压减低而不伴有心率减慢效应,而高浓度的APⅢ对血压和心率均有抑制效应。这些结果提示,ANP可能具有抑制延髓交感中枢作用并可能作为动脉压力感受器中枢通路的一种化学递质或调质。     

大鼠延髓腹面尾端加压区与头端加压区心血管效应的差异

雄性Ｗｉｓｔａｒ大鼠５６只,乌拉坦麻醉,双侧脑区微量注射谷氨酸钠,观察延髓腹面尾端加压区（ｃＶＭＰ）与头端加压区（ｒＶＭＰ）心血管效应的差异。结果表明：ｃＶＭＰ的升压效应弱于ｒＶＭＰ,不伴随明显的心率效应,ｒＶＭＰ升压效应强于ｃＶＭＰ兼有明显的心率增加。在谷氨酸钠兴奋作用下,ｒＶＭＰ抑制动脉压力感受反射,而ｃＶＭＰ易化该反射。提示两者对心血管功能的影响具有不同的神经途径和机能意义。     

内皮素通过最后区易化大鼠延髓腹外侧头端区神经元活动

在３５只切断双侧缓冲神经、用氨基甲酸乙酯－α氯醛糖混合麻醉的Ｓｐｒａｇｕｅ－Ｄａｗｌｅｙ大鼠,应用细胞外记录的电生理学方法,由ＲＭ－６０００型多道生理记录仪和ＷＳ－６８２Ｇ热阵记录器（频响范围０～２．８ｋＨｚ）同步记录血压、心率和单位神经元放电,观察颈动脉注射内皮素对８７个延髓腹是头端区（ＲＶＬＭ）自发放电神经元活动的影响,所得结果如下;（１）颈动脉注射ＥＴ－１（０．３ｎｍｏｌ／ｋｇ）时３６个单位     

延髓腹外侧区在降压反射中的作和

在各种心血管反射中,降压反射是最主要的,延髓腹外侧区在降压反射中起重要作用,目前认为降压反射中枢通路中至少有四种成分是最基本的：（１）孤束核中的神经元;（２）延髓腹外侧头端的交感前运动细胞;（３）延髓腹外侧尾端;（４）疑核或／和迷走神经背运动核。此外,兴奋性与抑制性氨基酸受体和抑制性神经元也是中枢通路的关键成分,下丘脑视上核与室旁核的升压素分泌细胞也有一定作用。     

延髓腹外侧区在降压反射中的作用

在各种心血管反射中,降压反射是最主要的,延髓腹外侧区在降压反射中起重要作用。目前认为降压反射中枢通路中至少有四种成分是最基本的：（１）孤束核中的神经元;（２）延髓腹外侧头端的交感前运动细胞;（３）延髓腹外侧尾端;（４）疑核或和迷走神经背运动核。此外,兴奋性与抑制性氨基酸受体和抑制性神经元也是中枢通路的关键成分。下丘脑视上核与室旁核的升压素分泌细胞也有一定作用     

乙酰胆碱对离体大鼠延髓头端腹外侧区神经元自发放电的影响

在８９张Ｓｐｒａｇｕｅ－Ｄａｗｌｅｙ大鼠延髓脑片,用玻璃微电极记录到１６５个延髓头端腹外侧区神经元的自发放电,其放电形式有三种：规则型;不规则型,静息型,乙酰胆碱对自发放电有兴奋,抑制双相和无影响四种效应,各占所测试神经元数的４１．８％,２０％,３％和自发放电有兴奋,抑制双相和无双相和影响四种效应,各占所有测试神经元数的４１．８％,２０％,３Ｔ和３５．２％。     

Involvement of Nitric Oxide in the Medullary Control of Circulation in Normotensive Rats

In acute experiments on anesthetized (urethane) normotensive rats, we studied the hemodynamic effects of unilateral microinjections of a nitric oxide (NO) donor, sodium nitroprusside, into the medullary nuclei participating in central cardiovascular control. We studied also the effects of modulation of the intensity of NO production: enhancing its synthesis by intramedullary injections of exogenous L-arginine or inhibiting this process with an inhibitor of neuronal NO synthase (nNOS), L-NNA, or with an inhibitor of arginase, norvaline. Intramedullary injections of the above agents were confined to the nucleus of the tractus solitarius, dorsal motor nucleus of the vagus, nucleus ambiguous, and lateral reticular nucleus. We tried to evaluate the possibility of production of NO from L-arginine in central neurons of normotensive rats via not only the well-known NO synthase pathway, but also via an alternative arginase-mediated pathway of metabolism of the above amino acid. Our results demonstrated that both enzymes are potentially active: injections of the mentioned inhibitors of the enzymes into the medullary neuronal structures induced marked shifts in the systemic arterial pressure (SAP), the integrative parameter characterizing the state of the cardiovascular system. After preliminary administration of an nNOS inhibitor, 7-nitroindazole (30 mg/kg, i.p.) or an inhibitor of arginase, norvaline (2 g, i.v.), injections of L-arginine into the medullary nuclei failed to evoke significant shifts in the SAP. We suggest that the comparative degree of activation of nNOS or arginase in the medullary nuclei depends on different factors, first of all on the level of oxygenation of the nerve tissue. An inverse dependence is likely to exist between the levels of activation of the above enzymes.     

Effect of Endothelin on Vasomotor and Respiratory Neurons in the Rostral Ventrolateral Medulla in Rats

1. We have previously shown that intracisternal administration of endothelin-1 (ET-1) elicited cardiorespiratory responses acting on the ventral surface of the medulla oblongata (VSM) subjacent to the rostral ventrolateral medulla (RVLM). In this study, we examined whether vasomotor and respiratory neurons in RVLM participate in above-mentioned responses and whether those neurons respond to direct iontophoretic application of ET-1 and/or an ET-A receptor antagonist, FR139317.2. Unit activity of vasomotor, respiratory, or nociceptive neurons in RVLM was recorded together with arterial blood pressure (AP) and heart rate (HR) in urethane-anesthetized Sprague-Dawley rats.3. Intracisternal administration or topical application of ET-1 (0.1–1 pmol) to VSM caused excitation of the majority of vasomotor neurons (15/18) and respiratory neurons (10/11) but not in nociceptive neurons (0/7). Changes in neuronal activity were in similar time course with corresponding changes in AP and HR. Iontophoretic application of ET-1 to the vicinity of recording neuron caused excitation in 19 of 21 vasomotor neurons without affecting AP nor HR. Remaining two neurons were insensitive to ET-1. FR139317 did not affect basal activity of the vasomotor neurons but inhibited ET-1-evoked excitation. Twenty-four of 40 respiratory neurons were excited and 13 were inhibited by iontophoretic application of ET-1. Five of ET-1-excited respiratory neurons were inhibited by FR139317 alone while six of ET-1-inhibited neurons were not affected by FR139317 alone. In both cases, FR139317 inhibited the effect of simultaneously applied ET-1. Iontophoretic application of ET-1 excited only one out of 10 nociceptive neurons so far tested.4. These results support the view that intracisternally administered ET-1 alters activity of vasomotor and respiratory neurons in the RVLM, at least in part by acting directly on neurons themselves and hence causes systemic cardiorespiratory changes. Majority of vasomotor and respiratory neurons should express ET-A receptors and some respiratory neurons are under tonic excitatory control by ET-1.     

Modulatory Inputs on Sympathetic Neurons in the Rostral Ventrolateral Medulla in the Rat

1. The first part of this study looks at spontaneously active neurons located in the rostral ventrolateral medulla (RVLM) with projections to the thoracic spinal cord. Sixteen neurons were intracellularly recorded in vivo. Four out of 16 neurons were antidromically activated from the thoracic spinal cord (axonal conduction velocities varied from 1.8 m/s to 9.5 m/s).2. The simultaneous averages of the neuronal membrane potential and arterial blood pressure triggered by the pulsatile arterial wave or the EKG-R wave demonstrated changes in membrane potential (hyperpolarization or depolarization) locked to the cardiac cycle in four neurons in this group. These neurons (three of them bulbospinal) were further tested for barosensitivity by characterizing the responses to electrical stimulation of the aortic depressor nerve. Four neurons responded with inhibitory hyperpolarizing responses characterized as inhibitory postsynaptic potentials (IPSP) to aortic nerve stimulation (onset latency: 32.3 ± 5.0 ms; mean ± SEM).3. In two neurons in the RVLM, one of them characterized as barosensitive, electrical stimulation of the opposite RVLM (0.5 Hz, 1.0 ms pulse duration, 25–100 A) elicited excitatory postsynaptic potentials (EPSPs) with latencies of 9.07 and 10.5 ms. At resting membrane potential, the onset latency of the evoked EPSPs did not change with increasing stimulus intensities. Some of the recorded neurons were intracellularly labelled with biocytin for visualization. They were found in the RVLM.4. These experiments in vivo would support the idea of a functional commissural pathway between the RVLM of both sides.5. Anatomical data have shown that some of those commissural bundle fibers originate in the C1 adrenergic neuronal group in the RVLM. In the second part of this study, we used an intracellular recording technique in vitro to investigate the effects of the indirect adrenergic agonist tyramine on neurons in the RVLM with electrophysiological properties similar to premotor sympathetic neurons in vivo.6. Tyramine (0.5–1 mM) produced a pronounced inhibitory effect with hyperpolarization and increase in membrane input resistance on two neurons characterized as regularly firing (R), and on one neuron characterized as irregularly firing (I). This effect was preceded by a transient depolarization with increases in firing rate.7. These results would indicate that neurons in the RVLM recorded in vitro and with properties similar to premotor sympathetic neurons can be modulated by catecholamines released from terminals probably making synaptic contacts.     

Regulation of Proenkephalin Synthesis in Adrenal Medullary Chromaffin Cells

The synthesis of proenkephalin was assessed in primary cultures of bovine adrenal medullary chromaffin cells by incubation of the cells with [35S]methionine, digestion of proenkephalin-derived peptides with trypsin and carboxy-peptidase B, and quantitation of radioactivity incorporated into Met-enkephalin following reversed-phase HPLC. Nicotine, histamine, and vasoactive intestinal peptide each enhanced the rate of proenkephalin synthesis approximately 10-fold when examined between 16 and 32 h after the drug or hormone addition. Inclusion of nifedipine (1 microM) partially blocked the stimulatory effect of nicotine, but not that of vasoactive intestinal peptide or histamine, or proenkephalin synthesis. Theophylline, tetrabenazine, and angiotensin II also increased the rate of proenkephalin synthesis (three- to eight-fold). These increases in the apparent rate of proenkephalin synthesis were not attributable to altered [35S]methionine specific radioactivity or rates of turnover and did not reflect similar increases in total protein synthesis. The half-life for turnover of Met-enkephalin sequences was 3-4 days in the cultured chromaffin cell. These studies directly show that proenkephalin synthesis is the primary regulatory step in control of chromaffin cell opioid peptide content.     

Chemoreception and feeding behaviour in a caterpillar: towards a model of brain functioning in insects

The quantitative relationship between chemoreceptor activity during food intake and total amount of food ingested per unit of time has been studied in last instar larvae of Pieris brassicae. Data on sensory responses and feeding intensity, derived from a study by Blom (1978), show a strong correlation between receptor activity during 1 sec periods of stimulation and food intake during 24 h. It is concluded that nerve impulses from chemoreceptors, which signal the presence of different phagostimulants, are summated algebraically in the central nervous system. Impulses from feeding deterrent receptors counteract the effects of phagostimulants. One impulse from a deterrent receptor neutralizes 2.5 impulses from phagostimulant receptors. Based on the relationships observed a simple model of a feeding centre in the central nervous system has been constructed (Fig. 8).     

Metabolic Pools of ATP in Cultured Bovine Adrenal Medullary Chromaffin Cells

Cultured bovine adrenal chromaffin cells contain a pool of ATP sequestered within the chromaffin vesicles and an extravesicular pool of ATP. In a previous study it was shown that the turnover of ATP in the extravesicular pool was biphasic. One phase occurred with a t1/2 of 3.5-4.5 h whereas the second phase occurred with a t1/2 of several days. The studies described here were undertaken to characterize further the vesicular and extravesicular pools of ATP by examining the effects of metabolic inhibitors, adenosine, and digitonin on ATP utilization and subcellular localization immediately after and 48 h after labeling with [3H]adenosine and 32Pi. Immediately after labeling a combination of cyanide, 2-deoxy-D-glucose, the beta-glucono-1,5-lactone resulted in a 90-95% depletion of the labeled ATP but only a 25% depletion of the endogenous ATP within 30 min. Forty-eight hours after labeling, addition of the inhibitors resulted in a 70% depletion of the [3H]ATP but only a 25% depletion of the [32P]ATP and endogenous ATP. Addition of 10 microM adenosine to the media resulted in a similar loss of [3H]ATP in cells examined immediately after or 48 h after labeling. Adenosine increased the amounts of [32P]ATP when added immediately after labeling but had no effect on the [32P]ATP content when added 48 h after labeling.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of Reserpine and Tetrabenazine on Catecholamine and ATP Storage in Cultured Bovine Adrenal Medullary Chromaffin Cells

The in vivo storage relationship between catecholamines and ATP in chromaffin vesicles of cultured bovine adrenal medulla cells was investigated using drugs that block vesicular catecholamine uptake. Three-day treatments with reserpine and tetrabenazine causing 85-90% depletion of catecholamines resulted in 41-46% reductions in cellular ATP content. Subcellular fractionation of reserpine-treated cells indicated that the ATP is lost from the chromaffin vesicle pool. This was confirmed in experiments using metabolic inhibitors to differentiate the vesicular and extravesicular ATP pools. The vesicular ATP loss was not proportional to that of catecholamines, resulting in a reduction by 50% in the chromaffin vesicle mole ratio of catecholamines to ATP after 48 h of treatment. In metabolic labeling studies, it was found that reserpine treatment reduced the incorporation of [3H]adenosine into vesicular ATP selectively, but it reduced the incorporation of 32Pi into both the vesicular and extravesicular pools. The reduction of the [3H]adenosine incorporation was not due to diminished vesicular nucleotide uptake resulting from low catecholamine levels, because when the catecholamines were depleted by tetrabenazine pretreatment followed by removal of the drug before labeling, no reduction in [3H]adenosine incorporation was observed. When present during the labeling, tetrabenazine was found to be a reversible inhibitor of plasma membrane adenosine uptake. The observed loss of adenine nucleotides from catecholamine-depleted chromaffin vesicles in vivo provides evidence that interactions between ATP and catecholamines are important in the vesicular storage of high concentration of these compounds.     

Central Regulation of Adrenal Tyrosine Hydroxylase: Effect of Induction on Catecholamine Levels in the Adrenal Medulla and Plasma

The effect of induction of adrenal tyrosine hydroxylase (TH) by various centrally acting drugs on catecholamine levels in adrenal and plasma was investigated in rats. All the drugs tested, namely oxotremorine, Piribedil, B-HT 920, and HA-966, produced significant increases in adrenal dopamine content and plasma epinephrine level. Denervation of the adrenal abolished the increase in adrenal dopamine as it did the induction of tyrosine hydroxylase. The results suggest that the induced increase of adrenal TH activity, as mediated by certain drugs, results in an elevation of the plasma epinephrine level and that the adrenal dopamine content is a better indicator of the catecholamine-synthesizing capacity of the adrenal medulla than are the other catecholamines.     

Turnover and Storage of Newly Synthesized Adenine Nucleotides in Bovine Adrenal Medullary Cell Cultures

The adenine nucleotide stores of cultured adrenal medullary cells were radiolabeled by incubating the cells with 32Pi and [3H]adenosine and the turnover, subcellular distribution, and secretion of the nucleotides were examined. ATP represented 84-88% of the labeled adenine nucleotides, ADP 11-13%, and AMP 1-3%. The turnover of 32P-adenine nucleotides and 3H-nucleotides was biphasic and virtually identical; there was an initial fast phase with a t1/2 of 3.5-4.5 h and a slow phase with a half-life varying from 7 to 17 days, depending upon the particular cell preparation. The t1/2 of the slow phase for labeled adenine nucleotides was the same as that for the turnover of labeled catecholamines. The subcellular distribution of labeled adenine nucleotides provides evidence that there are at least two pools of adenine nucleotides which make up the component with the long half-life. One pool, which contains the bulk of endogenous nucleotides (75% of the total), is present within the chromaffin vesicles; the subcellular localization of the second pool has not been identified. The studies also show that [3H]ATP and [32P]ATP are distributed differently within the cell; 3 days after labeling 75% of the [32P]ATP was present in chromaffin vesicles while only 35% of the [3H]ATP was present in chromaffin vesicles. Evidence for two pools of ATP with long half-lives and for the differential distribution of [32P]ATP and [3H]ATP was also obtained from secretion studies. Stimulation of cell cultures with nicotine or scorpion venom 24 h after labeling with [3H]adenosine and 32Pi released relatively twice as much catecholamine as 32P-labeled compounds and relatively three times as much catecholamine as 3H-labeled compounds.     

Effects of Lead Ions on Events Associated with Exocytosis in Isolated Bovine Adrenal Medullary Cells

Lead buffers (citrate and Tiron) were used to investigate the effects of low concentrations (0.1-6 microM) of Pb2+ on stimulus-secretion coupling in isolated bovine chromaffin cells. Nicotinic agonists and high K elicit secretion by enhancing Ca2+ influx into chromaffin cells. Pb2+ inhibited the catecholamine secretion in response to 500 microM carbachol and 77 mM K+ depolarization but was without significant effect on basal secretion. Pb2+ also inhibited the influx of 45Ca occurring in response to these agents. The K0.5 values for inhibition suggest that the carbachol-evoked flux is more sensitive to Pb2+ than influx in response to a direct depolarization. When extracellular calcium was lowered in the absence of Pb2+, both secretion and 45Ca entry were reduced. The effects of Pb2+ were comparable to those of lowered Ca2+. 22Na influx through nicotinic receptor-mediated channels, measured in the presence of tetrodotoxin (2 microM) and ouabain (50 microM), was inhibited by Pb2+. The results suggest that Pb2+ inhibits exocytotic catecholamine secretion by inhibiting Ca2+ influx. The differential sensitivity to Pb2+ of K- and carbachol-evoked 45Ca flux, coupled with the 22Na measurements, indicates that Pb2+ inhibits the movement of ions through acetylcholine-induced channels as well as through voltage-sensitive calcium channels.