豚鼠在体肠纱膜下神经节细胞兴奋的来源

本文运用在体细胞内记录法,观察了与肠系膜下神经节（ＩＭＧ）相连的四组神经对ＩＭＧ神经元电位活动的影响。结果显示切断或阻滞任一组神经均使ＩＭＧ细胞电活动受抑。其中结肠神经（ＣＶ）和腹下神经（ＨＮ）分别传导源自结肠尾段和膀胱等盆腔脏器的外周性兴奋,节间神经（ＭＮ）同时传导源自脊髓的中枢性和结肠的外周性兴奋。定量研究表明外周比中枢的影响更重要。因此ＩＭＧ不仅是传统认为的“信息传递站”,而且对中枢和外周     

豚鼠在体肠系膜下神经节细胞兴奋的来源

本文运用在体细胞内记录法,观察了与肠系膜下神经节（ＩＭＧ）相连的四组神经对ＩＭＧ神经元电位活动的影响。结果显示切断或阻滞任一组神经均使ＩＭＧ细胞电活动受抑。其中结肠神经（ＣＮ）和腹下神经（ＨＮ）分别传导源自结肠尾段和膀胱等盆腔脏器的外周性兴奋,节间神经（ＡＭＮ）同时传导源自脊髓的中枢性和结肠的外周性兴奋。定量研究表明外周比中枢的影晌更重要。因此ＩＭＧ不仅是传统认为的“信息传递站”,而且对中枢和外周信息有整合作用。     

豚鼠肠系膜下神经节细胞电活动与结肠运动的关系

在豚鼠肠系膜下神经节(IMG)及其支配的结肠段联合标本上,对IMG细胞内电位与肠段纵肌或环肌舒缩活动进行了同步记录。实验结果表明:(1)肠段预置张力为零时,约50％IMG细胞有自发的快兴奋性突触后电位(EPSP)活动,切断结肠神经或以筒箭毒(50μmol/L)灌流IMG后消失;(2)筒箭毒或低钙高镁溶液阻断神经节传递时,环肌节律性收缩幅度增大,节律变慢,但对纵肌节律性收缩无明显影响,(3)串刺激节前神经,在IMG细胞引起一串快EPSP或动作电位并常跟随迟慢的EPSP,同时,纵肌在0.1-0.2s潜伏期后出现迅速的、时程基本与动作电位串一致的舒张波,后者在筒箭毒灌流IMG后消失,而环肌运动可见舒张、舒张波延长或收缩波增大。结果提示:IMG不仅中继经典的胆碱能传出功能,还参与以胆碱能传递为中介的肠-肠反射,该反射活动的传出效应主要在于抑制环肌收缩。     

铃蟾肽介导的豚鼠肠系膜下神经节非胆碱能迟慢兴奋性突触后电位

应用细胞内记录技术,对铃蟾肽(bombesin,BOM)在豚鼠离体肠系膜下神经节(inferior mesenteric ganglion,IMG)非胆碱能兴奋性突触传递中的作用进行了研究。重复电刺激突触前结肠神经,有74．3％(52／70)IMG细胞可诱发迟慢兴奋性突触后电位(ls-EPSP)。在可引出ls-EPSP的细胞中,22％(4／18)细胞同时对BOM和SP敏感。用BOM持续灌流IMG,可明显抑制对BOM敏感细胞的ls-EPSP,对BOM不敏感细胞的ls-EPSP则无影响,且BOM受体与SP受体间无交叉脱敏。BOM受体阻断剂tyr^4[D-phe^12]bombesin能明显可逆性地抑制BOM敏感细胞的ls-EPSP和去极化,但对BOM不敏感细胞则无影响。研究结果提示,BOM可能是介导豚鼠IMG细胞ls-EPSP的一种递质。     

5—羟色胺介导的豚鼠肠系膜下神经节突触传递

本实验通过豚鼠离体肠系膜下神经节(IMG)的细胞内生物电记录方法观察到:(1)5-羟色胺(5-HT 1-100μmol/L)灌流可在部分 IMG 细胞引起与非胆碱能迟慢兴奋性突触后电位(Is-EPSP)相似的缓慢去极化;(2)持续灌流5-HT 可使对5-HT 敏感的 IMG 细胞的Is-EPSP 明显阻抑;(3)5-HT 去极化及5-HT 敏感细胞的 Is-EPSP 均可为5-HT 再摄取抑制剂氟苯氧丙胺(50μmol/L)所增大,而对5-HT 不敏感细胞的 Is-EPSP 则不受这种药物的影响,(4)5-HT 合成抑制剂对氯苯丙氨酸(PCPA)预处理可使 IMG 细胞的 Is-EPSP 的出现率和去极幅度均明显减低。上述结果表明:5-HT 可能参与介导豚鼠部分 IMG 细胞的Is-EPSP。     

豚鼠交感神经节非胆碱能迟慢兴奋性突触后电位与蛙皮素、P物质的关系

运用玻璃微电极细胞内记录技术,观察豚鼠(Cavia porcellus)离体肠系膜下神经节(IMG)细胞非胆碱能迟慢兴奋性突触后电位(Is—EPSP)与蛙皮素(BOM)、P物质(SP)的关系,以探讨肽类神经递质在外周神经系统中的作用。结果显示,SP去极化、BOM去极化与Is—EPSP具有相关性;SP受体脱敏使SP敏感细胞的Is—EPSP减弱或消失,但不影响BOM引起的去极化;BOM受体脱敏使BOM敏感细胞的Is—EPSP减弱或消失,但不影响SP引起的去极化。大部分Is—EPSP阳性细胞对SP、BOM敏感,而对SP、BOM均不敏感的细胞多数不出现Is—EPSP。结果提示,BOM、SP通过IMG细胞膜上相应受体参与了Is-EPSP的形成,受体间无交互脱敏现象。     

蛙皮素对豚鼠肠系膜下神经节细胞的生物电影响

应用离休细胞内记录技术,观察了蛙皮素（BOM）对豚鼠离体肠系膜下神经节（IMG）细胞膜电位和膜电阻的影响,结果表明,181个IMG细胞在压力注射BOM（10^-5mol/L,1-15pulse,3-15ms)时呈现缓慢去极化（84．0％）,先超极化后去极化（8．3％）,和无明显反应（7．7％）,在10个细胞上灌流BOM（10^-7-10^-6mol/L,60s),90％的细胞亦缓慢去极化,该去极化反应受低钙／高镁溶液的影响,但不为胆碱和肾上腺素受体阻断剂所阻断;膜电阻表现为减小（60．0％）,不变（35．0％）和增大（5．0％）,说明BOM可能存在于豚鼠IMG细胞上且发挥易化作用。     

大鼠脊髓诱发电位与脊髓损伤的关系——Ⅲ.脊髓局部损毁后电位的变化及电位来源分析

本文描述了大鼠脊髓L_1节段后柱、后索、侧索和前角的诱发电位及其损伤后的变化,并观察了切断L_4、L_5脊神经背、腹根与横断高位颈髓对电位的影响,以进行行电位来源分析。结果可见,上述四个区域的诱发电位基本由早反应三相波和晚反应组成。分别电解损毁这些部位后,电位波幅均普遍降低,晚期反应较早反应降低明显。后柱或后索受损对电位影响最大。局部损毁后可见L_1及T_(13)水平的硬膜上电位改变明显,尤其晚反应减弱、波峰平坦。反应时值与潜伏时未见明显改变。切断L_4脊神经背、腹根后、电位基本消失。去大脑对电位未见明显影响。结果表明,刺激坐骨神经诱发的脊髓电位起源于低位腰段传入神经和脊髓内多通路的兴奋传导,在一定程度上受腹根逆行活动的影响,与大脑及脊髓下行传导束活动无直接联系。脊髓诱发电位的幅度与波形改变可作为脊髓损伤的判断指标之一。     

辣椒素对大鼠肠系膜下神经诱发动作电位的影响

目的:探讨辣椒素对肠系膜下神经诱发动作电位的影响。方法:对大鼠肠系膜下神经节施加不同浓度(分别为2.5、5、10g/L)的辣椒素或其载体(对照)后,在其中枢端给予能引起反应的方波刺激,记录节后神经外周端的动作电位。结果:虽然实验动物存在个体差异,且辣椒素对其神经作用的阈值也有所不同,但在大多数情况下,当辣椒素浓度为2.5～5g/L、作用3min后,即可使神经的敏感性降低;并表现出了较为明显的剂量相关性,即随着辣椒素浓度的升高,其对神经的脱敏作用也逐渐增强。结论:辣椒素对肠系膜下神经节内的交感和副交感神经均具有抑制作用。     

兔肠系膜下神经节细胞的两种非胆碱能性慢突触后电位

以常规细胞内记录技术对兔肠系膜下神经节细胞的跨膜电位进行了观察。对节前神经的短串脉冲刺激,可诱发出一串快兴奋性突触后电位(f-EPSP)或顺向动作电位;在此之后,大多数细胞还出现一个持续约2min 的缓慢去极化电位。该电位具有抗箭毒和阿托品性质,受低钙高镁溶液的可逆性阻抑,因而可称为非胆碱能性兴奋性突触后电位,或者也可归入迟慢兴奋性突触后电位(ls-EPSP)。多数细胞的 ls-EPSP 伴有膜电阻增大,电位的幅度随细胞静息电位的超极化而变小;提示在这些细胞上,钾电导的失活很可能参与了电位的发生。以P物质溶液灌流神经节未见该电位有显著改变。另外,在箭毒化加阿托品化的神经节中,还发现少数细胞对节前神经的串刺激发生一个持续约一分钟的超极化电位。它也具有抗胆碱能受体阻断剂的性质,受低钙高镁溶液可逆性阻抑,为此我们命之为“极慢抑制性突触后电位”(vs-IPSP),以区别于“慢抑制性突触后电位”(s-IPSP),后者是通常用以表示一种胆碱能性的慢电位。本文所述的这两种非胆碱能性的突触电位有关递质,尚待探索。     

Spinal and supraspinal contributions to central sensitization in peripheral neuropathy

We will focus on spinal cord dorsal horn lamina I projection neurones, their supraspinal targets and involvement in pain processing. These spinal cord neurons respond to tonic peripheral inputs by wind-up and other intrinsic mechanisms that cause central hyper-excitability, which in turn can further enhance afferent inputs. We describe here another hierarchy of excitation - as inputs arrive in lamina I, neurones rapidly inform the parabrachial area (PBA) and periaqueductal grey (PAG), areas associated with the affective and autonomic responses to pain. In addition, PBA can connect to areas of the brainstem that send descending projections down to the spinal cord - establishing a loop. The serotonin receptor, 5HT3, in the spinal cord mediates excitatory descending inputs from the brainstem. These descending excitatory inputs are needed for the full coding of polymodal peripheral inputs from spinal neurons and are enhanced after nerve injury. Furthermore, activity in this serotonergic system can determine the actions of gabapentin (GBP) that is widely used in the treatment of neuropathic pain. Thus, a hierarchy of separate, but interacting excitatory systems exist at peripheral, spinal and supraspinal sites that all converge on spinal neurones. The reciprocal relations between pain, fear, anxiety and autonomic responses are likely to be subserved by these spinal-brainstem-spinal pathways we describe here. Understanding these pain pathways is a first step toward elucidating the complex links between pain and emotions.     

Tachykinins as modulators of the micturition reflex in the central and peripheral nervous system

In the normal urinary bladder, tachykinins (TKs) are expressed in a population of bladder nociceptors that is sensitive to the excitatory and desensitizing effects of capsaicin (i.e., capsaicin-sensitive primary afferent neurons (CSPANs)). Several endobiotics or xenobiotics excite CSPANs and release TKs and other mediators at both the peripheral and spinal cord level. The peripheral release of TKs determines a set of responses (known as neurogenic inflammation) that includes vasodilatation, plasma protein extravasation, smooth muscle contraction and stimulation of afferent nerves. Following chronic inflammation, both immune cells and capsaicin-resistant sensory neurons can de novo express TKs: whether these pools of TKs are releasable and contribute to inflammatory processes is presently unsettled. At the spinal cord level, the release of TKs contributes in determining an altered pattern of vesicourethral reflexes in response to nociceptive stimulation of the bladder by conveying: (a) the afferent transmission to supraspinal sites, and (b) descending or sensory inputs to the sacral parasympathetic nucleus (SPN). Recent evidence also attribute a synergetic role of TKs in the supraspinal modulation of the sensory arm of the micturition reflex.The overall available information suggests that TK receptor antagonists may affect bladder motility/reflexes which occur during different pathological states, while having little influence on the normal motor bladder function.     

Impaired Excitatory Drive to Spinal Gabaergic Neurons of Neuropathic Mice

Adequate pain sensitivity requires a delicate balance between excitation and inhibition in the dorsal horn of the spinal cord. This balance is severely impaired in neuropathy leading to enhanced pain sensations (hyperalgesia). The underlying mechanisms remain elusive. Here we explored the hypothesis that the excitatory drive to spinal GABAergic neurons might be impaired in neuropathic animals. Transgenic adult mice expressing EGFP under the promoter for GAD67 underwent either chronic constriction injury of the sciatic nerve or sham surgery. In transverse slices from lumbar spinal cord we performed whole-cell patch-clamp recordings from identified GABAergic neurons in lamina II. In neuropathic animals rates of mEPSC were reduced indicating diminished global excitatory input. This downregulation of excitatory drive required a rise in postsynaptic Ca²⁺. Neither the density and morphology of dendritic spines on GABAergic neurons nor the number of excitatory synapses contacting GABAergic neurons were affected by neuropathy. In contrast, paired-pulse ratio of Aδ- or C-fiber-evoked monosynaptic EPSCs following dorsal root stimulation was increased in neuropathic animals suggesting reduced neurotransmitter release from primary afferents. Our data indicate that peripheral neuropathy triggers Ca²⁺-dependent signaling pathways in spinal GABAergic neurons. This leads to a global downregulation of the excitatory drive to GABAergic neurons. The downregulation involves a presynaptic mechanism and also applies to the excitation of GABAergic neurons by presumably nociceptive Aδ- and C-fibers. This then leads to an inadequately low recruitment of inhibitory interneurons during nociception. We suggest that this previously unrecognized mechanism of impaired spinal inhibition contributes to hyperalgesia in neuropathy.     

Calcitonin gene-related peptide (CGRP) in capsaicin-sensitive substance P-immunoreactive sensory neurons in animals and man: distribution and release by capsaicin

The presence of calcitonin-gene related peptide (CGRP)-like immunoreactivity (-LI) in sensory neurons was established by immunohistochemistry and radioimmunoassay (RIA) in combination with high performance liquid chromatography (HPLC). CGRP-immunoreactive (-IR) nerve fibres were present in many peripheral organs including heart, ureter, uterus and gall bladder of guinea-pig and man. The distribution of CGRP-IR nerves in the dorsal horn of the spinal cord, of positive cell bodies in thoracic spinal and nodose ganglia and nerves in peripheral organs was closely related to that of substance P-LI. Double staining experiments revealed that in most cases peripheral CGRP-IR nerve terminals also contained SP-LI. However, different localization of SP- and CGRP-IR neurons was observed in the nucleus of the solitary tract as well as in the ventral horn of the spinal cord. In the heart, CGRP-IR nerves were associated with myocardial cells (mainly atria), coronary vessels, local parasympathetic ganglia as well as with the epi- and endocardia. Three to 4-fold higher levels of native CGRP-LI were observed in the atria than in the ventricles of the heart. HPLC analysis revealed that the major peak of CGRP-LI in the heart of rat and man had the same retention times as the synthetic equivalents. Systemic capsaicin pretreatment and adult guinea-pigs caused a loss of CGRP-IR terminals in the dorsal horn of the spinal cord as well as in peripheral organs including the heart. After capsaicin treatment, the content of CGRP-IR was reduced by 70% in the heart and by 60% in the dorsal part of the spinal cord. In superfusion experiments with slices from the rat spinal cord, a release of CGRP-LI was induced by 60 mM K+ and 3 microM capsaicin in a calcium-dependent manner.     

TRPV1 in GABAergic interneurons mediates neuropathic mechanical allodynia and disinhibition of the nociceptive circuitry in the spinal cord

Neuropathic pain and allodynia may arise from sensitization of central circuits. We report a mechanism of disinhibition-based central sensitization resulting from long-term depression (LTD) of GABAergic interneurons as a consequence of TRPV1 activation in the spinal cord. Intrathecal administration of TRPV1 agonists led to mechanical allodynia that was not dependent on peripheral TRPV1 neurons. TRPV1 was functionally expressed in GABAergic spinal interneurons and activation of spinal TRPV1 resulted in LTD of excitatory inputs and a reduction of inhibitory signaling to spinothalamic tract (STT) projection neurons. Mechanical hypersensitivity after peripheral nerve injury was attenuated in TRPV1(-/-) mice but not in mice lacking TRPV1-expressing peripheral neurons. Mechanical pain was reversed by a spinally applied TRPV1 antagonist while avoiding the hyperthermic side effect of systemic treatment. Our results demonstrate that spinal TRPV1 plays a critical role as a synaptic regulator and suggest the utility of central nervous system-specific TRPV1 antagonists for treating neuropathic pain.     

Canal and otolith inputs to single vestibular neurons in cats

Convergence of both afferents from the PC and saccular macula, and those from the PC and utricular macula on single vestibular neurons was noted by use of intercellular recording from vestibular neurons. Vestibular neurons were classified VO neurons (vestibulo-ocular proper neurons), VOS (Vestibulo-oculo-spinal neurons sending axon collaterals both to the extraocular motoneuron pools and to the spinal cord), VS neurons (vestibulospinal proper neurons) and V neurons (vestibular neurons without axons to the oculomotor nuclei or the spinal cord) on the basis of whether or not they responded antidromically to stimulation of the oculomotor nuclei and the spinal cord. Of the total 143 vestibular neurons recorded in the series of experiments on convergence of the PC and saccular afferents, 47 neurons (33%) were received inputs from both the PC and saccular nerves. Twenty-six of the 47 convergent neurons were identified as having the nature of VS neurons. Half (13/26) of those were activated monosynaptically from both the PC and saccular nerves. Only one saccular-activated neuron without PC inputs sent an axon to the oculomotor nuclei. In the other series of experiments on the convergence of the PC and utricular afferents, 41 (37%) of 111 vestibular neurons were proved to converge on inputs from both nerves. The majority (35/41) of the neurons received monosynaptic inputs from the PC nerve and polysynaptic EPSP-IPSP sequences from the utricular nerve, or vice versa. The ratio of PC-otolith convergent neurons among utricular-activated neurons (41/54, 76%) was higher than that among saccular activated neurons (47/88, 53%). The percentage of utricular alone neurons without PC inputs (13/111, 12%) was less than that of the saccular alone without PC inputs (41/145, 28%). In conclusion, the convergence of canal and otolith inputs likely contribute mainly to vestibulospinal reflexes including the vestibulocollic reflex, by sending inputs to the neck and other muscles during head inclination which creates the combined stimuli of angular and linear acceleration.     

Otolith and semicircular canal inputs to single vestibular neurons in cats.

Researchers studied the convergence of the vertical posterior semicircular canal (PC), saccular nerves (SAC), utricular nerves (UT), and horizontal semicircular canal nerves (HC) on single vestibular neurons. The vestibular neurons were categorized by their innervating targets. Vestibular neurons were classified as vestibulospinal proper neurons (VS), vestibulo-ocular proper neurons (VO), vestibulo-oculo-spinal neurons sending axon collaterals to the extraocular motoneuron pools and spinal cord (VOS), and vestibular nucleus neurons without axons to the oculomotor nuclei or the spinal cord (V). Results indicate that the percentage of convergence of VS neurons was higher that that of neurons sending axons to the oculomotor nuclei (VO and VOS). They conclude that the convergence of canal and otolith inputs likely contributes mainly to vestibulospinal reflexes by sending inputs to the neck and other muscles during head inclination, which creates the combined stimuli of angular and linear acceleration.     

Distribution of enteric nerve cells projecting to the superior and inferior mesenteric ganglia of the guinea-pig

Retrograde tracing, using Fast Blue dye, was employed to determine the distribution of enteric nerve cells that project to the superior mesenteric and inferior mesenteric ganglia of the guinea-pig. Retrogradely labelled neurons were found in the myenteric but not submucous ganglia. When the superior mesenteric ganglion was injected, labelled neurons were found in low frequencies (less than 5 nerve cell bodies/cm²) in the duodenum, jejunum, ileum, caecum and proximal colon. The distal colon was analysed in five segments of equal length (1–5; oral to anal). Segment 1 had about 4 labelled nerve cells/cm², whereas segments 2 to 5 displayed an average of about 25 nerve cells/cm². The rectum contained about 36 labelled neurons/cm². After injection of the inferior mesenteric ganglia with Fast Blue, no labelled neurons were found in the duodenum, jejunum, ileum or caecum. No labelled cells were observed in the gallbladder. A small number of labelled cells occurred in the proximal colon and in segment 1 of the distal colon. The frequency of labelled cells increased markedly in the more anal regions of the distal colon, and reached a peak in the rectum (138 cells/cm²). Both nerve lesions and immersion of the cut nerve in Fast Blue solution showed that the superior mesenteric nerve carries the axons of neurons located in the middle distal colon to the superior mesenteric ganglion. Almost half of the neurons in the rectum that project to the inferior mesenteric ganglia do so via the hypogastric nerves. Of neurons that projected to the inferior or superior mesenteric ganglia from the colon or rectum, similar proportions (about 75–80%) showed immunoreactivity for calbindin or VIP. For each of the prevertebral ganglia (coeliac, superior mesenteric and inferior mesenteric) the great majority of peripheral inputs arise from the large intestine.