扣带回前部内脏伤害感受神经元的生物电活动

为了从神经元水平探讨大脑皮层内脏伤害感受的特性及机制,应用玻璃微电极细胞内电位记录技术,研究18只猫扣带回前部312个神经元的自发生物电活动,及其对电刺激同侧内脏大神经的诱发反应.其中,82个为内脏伤害感受神经元,其自发生物电活动有5种主要形式.根据诱发反应的潜伏期等特性,内脏伤害感受神经元分为特异性内脏伤害感受神经元（76个,92.68％）和非特异性内脏伤害感受神经元（6个,7.32％）.内脏伤害性诱发反应分为兴奋性（65.86％）、抑制性（17.07％）及混合性反应（17.07％）3种.结果提示内脏大神经的传入通路投射到同侧扣带回前部;扣带回前部神经元具有内脏伤害感受作用,存有特异性与非特异性内脏伤害感受神经元,为痛觉特异性学说提供了新的实验依据.     

猫扣带回前部内脏伤害感受神经元的诱发反应

应用玻璃微电极细胞内电位记录技术,观察了２０史猫扣带回前部４６１个神经元对电刺激对侧内脏大神经的诱发反应及其电生理特性,在被观察的神经元中,１７６个为刺激相关神经元。根据诱发反应的特性,将其分为特异性内脏伤害感受神经元（１１４个,６４．７７％）、非特异性内脏伤害感受神经元（３４个,１９．３２％）及非内脏伤害感受神经元（２８个,１５．９１％）。诱发反应分为兴奋性（５９．４６％）、抑制性（２２．３０％     

猫皮层体感Ⅱ区内脏伤害感受神经元的电生理特性及形态特征

应用胞内记录和标记技术,观察了猫皮质第Ⅱ感觉区内脏大神经代表区的神经元对电刺激内脏大神经反应诱发反应及形态特征。结果表明,在２５１个记录单位中,有１０９个为内脏伤害性感受神经元,其诱发反应分为兴奋性、抑制性及混合性三类。在形式上ＩＳＰＳ及ＥＰＳＰ－ＩＰＳＰ序列反应较多。对其中２１个神经元用神经生物素进行细胞内电泳标记,显示细胞的形态特点是胞体较小,分布于皮质Ⅱ、Ⅲ、Ⅴ层,其中兴奋性和神经元形态多为     

猫扣带回前部内脏伤害感受神经元的膜电学特性

目的：研究猫扣带回前部内脏大神经刺激相关神经元的膜电生理特性,以便从神经元水平进一步了解大脑皮质内脏伤害感受的特性及机制,为痛觉理论“特异性学说”提供新的实验依据。方法：应用在体玻璃微电极细胞内电位记录技术及细胞内注入极化电流的方法,测量和计算神经元的膜电学参数。结果：将20只猫扣带回前部176个内脏大神经刺激相关神经元,分为内脏伤害(148个)和非伤害(28个)感受神经元。发现它们在膜电阻、时间常数、膜电容及I—V曲线等方面存在差异。注入去极化电流引发的放电幅值及频率也存在差异。结论：扣带回前部内脏伤害与非伤害感受神经元可能在细胞膜结构、细胞大小等形态学方面存有差别。     

猫皮层SⅠ区神经元对刺激VPL的反应及其膜电学特性的研究

本文采用细胞内记录技术,研究了猫皮层第一躯体感觉区（ｐｒｉｍａｒｙｓｏｍａｔｏｓｅｎｓｏｒｙｃｏｒｔｅｘａｒｅａ,ＳⅠ区）躯体伤害感受神经元膜的电学特性和对刺激腹后外侧核（ｖｅｎｔｒａｌｐｏｓｔｅｒｉｏｒｌａｔｅｒａｌｎｕｃｌｅｕｓ,ＶＰＬ核）的反应。极化电流绝对值小于或等于１０ｎＡ时,伤害感受神经元ＩＶ极相关（ｒ＝０９６）,整流作用不明显;极化电流绝对值大于１０ｎＡ时,在两个方向上发生整流,ＩＶ曲线表现为Ｓ型,其中伤害感受神经元的整流作用较非伤害感受神经元明显。伤害性感受神经元Ｒｍ、τ、Ｃｍ明显大于非伤害感受神经元（Ｐ＜００１或Ｐ＜００５）。刺激ＶＰＬ与刺激隐神经在ＳⅠ区伤害感受神经元的诱发反应中存有相似与不同两种形式。用细胞内电位记录方法证明了单一神经元有会聚现象。结果提示,ＳⅠ区伤害感受神经元与非伤害感受神经元可能在细胞膜形态结构、细胞体积大小等方面存在有意义的差别,从而反映其不同的生理功能。     

猫体感皮质内脏伤害感受神经元的膜电生理特性

应用细胞内电位记录技术,研究猫ＳＩ中内脏大神经皮质代表区的８５１个神经元膜电生理特性,用胞内注入极化电流的方法,测量和计算出神经元的膜电学参数,并进行内脏痛伤害感受及非相关神经元的电学特性比较,发现二者在膜电阻、时间常数、膜电容、静息电位、细胞活跃程度及神经元深度分布等方面存在差别。注电流引发的频率－电流、动作电位幅值－电流、及Ｉ－Ｖ曲线也有差异,结果提示ＳＩ区内脏伤害感受及非相关神经元在形态及膜结构上可能有不同之处,为痛觉特异学说提供资料。部分神经元用神经生物素进行细胞内电泳标记以显示功能细胞所在层次及形态,从单一皮层感受神经元的反应及形态特点探讨了内脏痛的感受特性。     

猫双侧扣带回前部内脏伤害感受神经元自发电活动特性的差异

本研究旨在阐明双侧大脑皮层扣带回前部(anterior cingulate gyrus,ACG)在内脏痛觉感受中的作用。实验用强电流刺激一侧内脏大神经(greater splanchnic nerve,GSN)作为内脏伤害性刺激,用在体微电极细胞内电位记录技术记录、分析ACG内神经元对刺激的反应和自发生物电活动。根据对GSN刺激是否发生反应,将ACG内神经元分为产生诱发反应的GSN刺激相关神经元(GSN-stimulus-relative neurons,GSRNs)与不产生诱发反应的非相关神经元。根据诱发反应的潜伏期等性质,GSRNs进一步分为内脏伤害感受神经元(visceral nociceptive neurons,VNNs)与非伤害感受神经元(non-visceral nocicep-tive neurons,NVNNs);而VNNs又分为特异性(specific visceronociceptive neurons,SVNNs)与非特异性内脏伤害感受神经元(non-specific visceronociceptive neurons,NSVNNs)。结果显示,对侧ACG内GSRNs比例为38.18%,显著高于同侧ACG内GSRNs比例(29.49%,P0.01),表明GSN传入纤维虽然投射到双侧ACG,但主要投射到对侧ACG。和同侧ACG相比,对侧ACG的SVNNs比例较低,而NSVNNs比例则较高(P0.01)。同侧ACG内GSRNs、VNNs、NVNNs和SVNNs的静息电位(restingpotential,RP)绝对值均显著低于对侧ACG的对应神经元(P0.01),而双侧ACG内NSVNNs的RP绝对值没有明显差异。双侧ACG内VNNs、NVNNs的自发生物电活动形式、自发放电频率和幅值之间无明显差别;VNNs中具有自发生物电活动的神经元比例显著高于NVNNs中的比例。以上结果提示双侧ACG皮层虽然都能感受GSN的伤害性刺激信号,但双侧ACG皮层的反应模式、反应程度不完全相同,从而为双侧脑功能的不对称性提供了新的科学实验依据。     

针刺过程中猴的操作式条件反射和大脑皮层体感区神经元电活动的观察

为了研究大脑皮层体感Ⅰ区与痛觉以及针刺镇痛作用的关系,以钾离子透入下肢皮肤作为痛刺激,以操作式条件反射作为痛指标,用钨丝微电极记录大脑皮层体感Ⅰ区单个神经元的电活动,在10只清醒、可以活动的猴子上进行了实验观察。在下肢的皮层代表区分离和记录了能对不同传入刺激起反应的神经元215个。28个对皮肤伤害性刺激起反应。其中25个发放增加(兴奋型神经元),3个发放减少(抑制型神经元)。对12个伤害性神经元进行了针刺效应的观察。在针刺过程中有6个神经元(4个兴奋型、2个抑制型)对伤害性刺激的反应受到抑制。其中有些神经元的抑制可伴有痛阈的升高,但两者未呈现严格的平行关系。皮层体感Ⅰ区中亦可记录到会聚性神经元,即同一神经元既可接受皮肤机械刺激,亦可接受皮肤伤害性刺激。其中有些神经元对于不同性质的刺激可以出现不同型式的反应,如对钾离子刺激皮肤出现抑制性反应而对触觉刺激出现兴奋性反应。针刺时,往往只有伤害性刺激引起的反应受到抑制,而机械刺激引起的反应没有明显变化。实验表明,针刺可以使皮层体感Ⅰ区的某些神经元对皮肤伤害性刺激的反应减弱;但是这种效应可能不是发生在皮层本身,而是续发于皮层下结构活动的变化。     

半胱胺对猫脊髓背角神经元伤害性热反应的抑制

在戊巴比妥钠麻醉和脊髓腰-1段全横切的16只猫上,观察生长抑素(somatostatin,SOM)的耗竭剂半胱胺对伤害性热刺激脚跖皮肤和电刺激胫后神经所引起的脊髓背角Ⅳ-Ⅵ层神经元单位反应的影响。静脉注射半胱胺50mg/kg对电刺激神经引起的伤害性反应无影响,100mg/kg可使被测试的13个神经元单位中的8个单位反应明显抑制。而静脉注射半胱胺50mg/kg可明显抑制伤害性热刺激所引起的脊髓背角神经元单位反应。用微电极将半胱胺微压注入背角胶质层也使背角神经元的伤害性热反应明显抑制,但只使13个单位中的7个单位对电刺激神经引起的伤害性反应轻度抑制。半胱胺对背角神经元伤害性反应的抑制可能由于耗竭了背角中的生长抑素。本文讨论了半胱胺对背角神经元伤害性热反应的抑制明显强于电刺激神经所诱发的伤害性反应的抑制的可能机制。     

会阴部皮肤和盆腔内脏的伤害感受性信息在猫骶髓后连合核神经元的汇聚

在戊巴比妥钠麻醉的猫上,用玻璃微电极细胞外记录的方法,观察了躯体和内脏的伤害性刺激对骶髓后连合核神经元活动的影响。结果表明,所有接受盆神经内Ａδ纤维传入的神经元皆为特异性伤害感受或广动力范围神经元－它们可被包括会阴部皮肤的躯体感受野的机械性及强电刺激诱发。上述结果提示,Ａδ纤维可能在盆腔内脏的伤害感受性传递中起重要作用。     

Difference in properties of spontaneous electric activities of visceral nociceptive neurons in bilateral anterior cingulate gyrus of cats

The aim of the present study is to explore the role of anterior cingulate gyrus (ACG) in bilateral cerebral cortex in visceral nociceptive sensation. Electrical stimulation of greater splanchnic nerve (GSN) was used as visceral nociceptive stimulus, and intracellular recording techniques in vivo was used to record and analyze the responses to stimuli and spontaneous electric activities of the neurons in the bilateral ACG. According to the responses to electrical stimulation of GSN, the neurons in the bilateral ACG were divided into GSN-stimulus-relative neurons (GSRNs) and GSN-stimulus-irrelative ones. According to the characteristics of the evoked responses to electrical stimulation of the GSN, GSRNs could be further classified into visceral nociceptive neurons (VNNs) and non-visceral nociceptive neurons (NVNNs). VNNs included specific visceral nociceptive neurons (SVNNs) and non-specific visceral nociceptive neurons (NSVNNs). The results showed that the percentage of GSRNs in the contralateral ACG (38.18%) was significantly higher than that in the ipsilateral ACG (29.49%, P<0.01), suggesting although GSN afferent fibers project to bilateral ACG, they mainly project to the contralateral ACG. Compared with ipsilateral ACG, contralateral ACG possessed lower proportion of SVNNs and higher proportion of NSVNNs (P<0.01). The absolute values of resting potentials (RP) of GSRNs, VNNs, NVNNs and SVNNs in ipsilateral ACG were less than those of corresponding neurons in contralateral ACG. However, there were no significant differences in the absolute values of RP of NSVNNs between ipsilateral and contralateral ACG. There were no significant differences in modes, frequencies and amplitudes of spontaneous electric activities of VNNs and NVNNs between ipsilateral and contralateral ACG. Additionally, the percentage of neurons having spontaneous electric activities from VNNs was significantly higher than that from NVNNs, which indicated that the excitability of VNNs was higher than that of the NVNNs in bilateral ACG. These results suggest that the patterns and degrees of the responses to nociceptive GSN-stimulation of the ipsilateral and contralateral ACG are different, thus providing new experimental data for the asymmetry of functions of the bilateral brain.     

Effects of abdominal or cardiopulmonary sympathetic afferents on upper cervical inspiratory neurons

Responses of upper cervical inspiratory neurons (UCINs) to abdominal visceral or cardiopulmonary sympathetic stimulation were studied using extracellular recordings from 213 UCINs in 54 pentobarbital sodium-anesthetized and paralyzed rats. Phrenic nerve activity was used to assess inspiration. The UCINs discharging during inspiration only were mainly in the C(1) segment, whereas phase-spanning UCINs were mostly in the C(2) segment. Phase-spanning activity was typically retained after overventilation or vagotomy. When greater splanchnic nerve (GSN) or cardiopulmonary sympathetic afferent (CPSA) fibers were electrically stimulated, augmented UCIN activity was observed in 65% of cells responding to CPSA stimulation but in only 17% of cells responding to GSN. Response latencies were 10.7 +/- 0.5 and 20.6 +/- 1.5 (SE) ms, respectively. Many augmented responses to CPSA stimulation (64%) and all augmented responses to GSN stimulation were followed by suppression of UCIN discharge (biphasic response). Phrenic nerve activity was suppressed by both GSN and CPSA stimulation, but with shorter latency for the latter (29 +/- 0.7 vs. 14.0 +/- 0.7 ms). Excitation of UCINs using CPSA stimulation occurs more often and by a more direct pathway than for GSN input.     

Dissociation of μ- and δ-opioid inhibition of glutamatergic synaptic transmission in superficial dorsal horn

Background

Although it has been widely accepted that the primary somatosensory (SI) cortex plays an important role in pain perception, it still remains unclear how the nociceptive mechanisms of synaptic transmission occur at the single neuron level. The aim of the present study was to examine whether noxious stimulation applied to the orofacial area evokes the synaptic response of SI neurons in urethane-anesthetized rats using an in vivo patch-clamp technique.

Results

In vivo whole-cell current-clamp recordings were performed in rat SI neurons (layers III-IV). Twenty-seven out of 63 neurons were identified in the mechanical receptive field of the orofacial area (36 neurons showed no receptive field) and they were classified as non-nociceptive (low-threshold mechanoreceptive; 6/27, 22%) and nociceptive neurons. Nociceptive neurons were further divided into wide-dynamic range neurons (3/27, 11%) and nociceptive-specific neurons (18/27, 67%). In the majority of these neurons, a proportion of the excitatory postsynaptic potentials (EPSPs) reached the threshold, and then generated random discharges of action potentials. Noxious mechanical stimuli applied to the receptive field elicited a discharge of action potentials on the barrage of EPSPs. In the case of noxious chemical stimulation applied as mustard oil to the orofacial area, the membrane potential shifted depolarization and the rate of spontaneous discharges gradually increased as did the noxious pinch-evoked discharge rates, which were usually associated with potentiated EPSP amplitudes.

Conclusions

The present study provides evidence that SI neurons in deep layers III-V respond to the temporal summation of EPSPs due to noxious mechanical and chemical stimulation applied to the orofacial area and that these neurons may contribute to the processing of nociceptive information, including hyperalgesia.     

束旁核、中央中核对皮层体感觉区伤害性反应单位的影响

实验在清醒麻痹的猫上进行。先用记录诱发电位的方法,证明大脑皮层第 I 体感觉区(SI区)同一部位可接受来自外周神经、丘脑束旁核(Pf)和中央中核(CM)的上传信号;然后在这一皮层部位寻找在强电流刺激腓总神经时出现伤害性反应的神经元。在所观察的24个单位中,有20个单位如果先给予 Pf 刺激,可看到上述伤害性反应明显增强,增强的百分率平均在90.0%以上;在这同一组皮层单位,如果预先刺激 CM,有21个单位的伤害性反应明显减弱,反应抑制都在-55.6%以上。Pf 刺激本身对所观察单位的自发放电影响不明显,但 CM 刺激可明显抑制它们的自发放电。     

刺激杏仁基底外侧核对外侧缰核神经元单位放电的影响

用玻璃微电极细胞外记录大鼠外侧缰核(LHN)神经元的单位放电。共记录了110个神经元。其中痛兴奋神经元(LHPE)75个;痛抑制神经元(LHPI)11个;广动力型神经元2个;无反应神经元17个;此外还有5个对躯体与内脏伤害性刺激反应不同的神经元。电刺激杏仁基底外侧核(以下简称杏仁核,AMG)对LHPE和LHPI的自发放电主要产生抑制作用,分别占总数的81.1％和72.7％,并抑制其对伤害性刺激的反应;对无反应神经元和广动力型神经元无明显影响。AMG内微量注射吗啡能抑制LHPE的伤害性刺激反应,但对其自发放电无明显影响。微量注射纳洛酮则可增加LHPE的自发放电频率,并加强其对伤害性刺激的反应。注射纳洛酮还可以取消电针对LHPE的伤害性刺激反应的抑制作用。     

刺激大鼠尾核对丘脑束旁核躯体-内脏会聚神经元放电的影响

实验记录大鼠丘脑束旁核躯体-内脏会聚(PfSV)神经元伤害性放电。观察刺激尾核(Cd)对 PfSV 神经元放电的影响。(1)Cd 对刺激内脏大神经诱发 PfSV 神经元伤害性放电有抑制作用(n=19)。(2)Cd 对刺激腓浅神经和内脏大神经诱发同一 PfSV 神经元伤害性放电均有抑制作用(n=11)。结果提示,躯体和内脏痛觉信息可会聚到丘脑束旁核同一神经元,Cd 可能不仅能抑制躯体痛也能抑制内脏痛。