大鼠延髓后角神经降压肽(NT)的亚细胞定位和胞吐释放

神经降压肽(NT)广泛分布于哺乳动物的中枢神经系统,具有明显的镇痛作用,为了探索其镇痛机理的形态学基础,本文应用电镜免疫组化技术,对大鼠延髓后角NT的超微结构和胞吐释放进行了研究。超微结构显示延髓后角浅层NT轴突终末形态多样,大小不一,含有圆形或多形性清亮小泡及数量不等的大颗粒小泡,它们主要与未标记的树突形成轴-树突触,其突触后成分有的还含有少量清亮小泡。NT免疫反应阳性树突可分为两类:一类主要含微管;另一类主要含大颗粒小泡,有的尚可见少量清亮小泡。这两类NT树突可成为未标记的含圆形小泡终末、多形性小泡终末以及突触小球中央轴突终末的突触后成分,提示后角浅层NT神经元可接受不同种类轴突终末(包括一级伤害性传入纤维)的传入(?)动,然后可能再通过一个抑制性中间神经元,抑制痛觉的传递。本文还观察到有少量NT终末内的大颗粒小泡靠近突触活性区处,而更多见它们沿非突触部位轴膜分布,并与其融合,形成胞吐。本文认为NT既可在突触活性区处又可能在非突触部位释放。     

大鼠三叉神经脊束核尾侧亚核胶状质非小球突触成分

用透射电镜观察了大鼠三叉神经脊束核尾侧亚核胶状质神经毡非小球的突触成分。非小球的突触大部分为轴树突触,此外还见到轴轴、树树及树轴突触。它们的轴突终末成分,按所含小泡的形状,区分为圆形小泡终末、扁平小泡终末、多形小泡终末及大颗粒小泡终末。圆形小泡终末根据小泡的大小又有大圆形小泡终末及小圆形小泡终末。本文还讨论了突触分类及各种轴突终末的机能意义。     

大鼠三叉神经脊束核Ⅱ尾侧亚核层内CB样、PV样阳性结构的分布及其突触联系

本研究用免疫组织化学方法观察了 Calbindin D-2 8k( CB)样和 Parvalbumin ( PV)样胞体、纤维和终末在三叉神经脊束核尾侧亚核 ( Vc) 层内的分布及它们的突触联系。在光镜下观察到 CB样和 PV样阳性胞体、纤维和终末在 II层内侧带 ( IIi)最为密集 ,PV样阳性神经元的胞体稍大 ,但数量少于 CB样阳性神经元。在电镜下观察到 CB样或 PV样阳性结构主要形成下列 4种突触联系 :( 1)阳性轴突终末与阳性或阴性轴突终末形成对称性轴 -轴突触和少量非对称性轴 -轴突触 ;( 2 )阳性轴突终末与阳性树突形成非对称性和对称性轴 -树突触 ;( 3 ) CB样阳性轴突终末与阴性树突主要形成非对称性轴 -树突触 ,PV样阳性轴突终末与阴性树突主要形成对称性轴 -树突触 ;( 4 )阴性轴突终末与阳性树突形成非对称性和对称性轴 -树突触。另外还可见到 CB样或PV样阳性或阴性树突、轴突及终末与 CB样、PV样阳性或阴性的初级传入纤维终末形成 型和 II型突触小球。 型突触小球数量较多 ,有典型的扇贝样初级传入纤维终末和不均一的小泡 ,线粒体少 ;II型突触小球的初级传入纤维终末粗大而清亮 ,外观不规则 ,有均匀一致的小泡和丰富的线粒体。根据上述结果可以推知在面口部伤害性信息的传递和调控过程中 ,Vc II层神经元发挥着重要的作用     

大鼠海马结构在空间辨别性学习记忆时的突触形态学观察

目的：探讨学习记忆活动是否可以引起大鼠海马结构发生突触可塑变化。方法：电子显微镜下对模型大鼠和对照大鼠的海马结构内突触复合体的形态进行了对比观察。结果：(1)模型大鼠海马CJ43区多形层内多为神经纤维，其中无髓纤维占多数，有髓纤维很少。在无髓纤维间见到形状不规则的轴突终末与周围的树突和树突棘分别形成轴—树突触和轴棘突触，后者多见。突触前膨大内充满圆形清亮、无核心的突触小泡和线粒体。突触后成分多为树突棘并在多处与突触前膜形成活性区，树突棘内见到棘器。(2)对照大鼠海马CA3区多形层内含有大量无髓纤维并见到较少的轴—树突触。该突触形状规则且活性带面积较小，突触后成分多为一个，突触小泡清亮、圆形无核心，偶见线粒体。(3)经Timm染色的两组大鼠海马CA3区多形层的超微结构特点是银颗粒仅沉积在苔藓纤维的髓鞘内和苔藓纤维的大终扣内，线粒体、树突干和树突棘内均无银颗粒。结论：正常生理活动和空间辨别性学习记忆活动均可引致大鼠海马CA3区多形层突触可塑性变化，但两者在形态上有所不同。     

大鼠三叉神经脊束核尾侧亚核Ⅱ层内CB样、PV样阳性结构的分布及其突触联系

本研究用免疫组织化学方法观察了CalbindinD-28k(CB)样和Parvalbumin(PV)样胞体、纤维和终末在三叉神经脊束核尾侧亚核(Vc)Ⅱ层内的分布及它们的突触联系.在光镜下观察到CB样和PV样阳性胞体、纤维和终末在Ⅱ层内侧带(Ⅱi)最为密集,PV样阳性神经元的胞体稍大,但数量少于CB样阳性神经元,在电镜下观察到CB样或PV样阳性结构主要形成下列4种突触联系:(1)阳性轴突终末与阳性或阴性轴突终末形成对称性轴-轴突触和少量非对称性轴-轴突触;(2)阳性轴突终末与阳性树突形成非对称性和对称性轴-树突触;(3)CB样阳性轴突终末与阴性树突主要形成非对称性轴-树突触,PV样阳性轴突转末与阴性树突主要形成对称性轴-树突触;(4)阴性轴突终末与阳性树突形成非对称性和对称性轴-树突触.另外还可见到CB样或PV样阳性或阴性树突、轴突及终末与CB样、PV样阳性或阴性的初级传入纤维终末形成Ⅰ型和Ⅱ型突触小球.Ⅰ型突触小球数量较多,有典型的扇贝样初级传入纤维终末和不均一的小泡,线粒体少;Ⅱ型突触小球的初级传入纤维终末粗大而清亮.外观不规则,有均匀一致的小泡和丰富的线粒体.根据上述结果可以推知在面口部伤害性信息的传递和调控过程中,VeⅡ层神经元发挥着重要的作用.     

猫孤束核胶状质亚核的突触构筑学研究

用透射电镜对猫的孤束核胶状质亚核(SNG)的突触型式进行了观察,除看到已报导的轴—树突触、轴—体突触、树—树突触外,还发现该核内含有轴—轴突触及突触球等结构。SNG内轴—树突触最常见,而轴—体突触、轴—轴突触和树—树突触则较少。各类突触中的突触囊泡多为圆形清亮囊泡,而扁平清亮囊泡和大颗粒囊泡较少。扁平清亮囊泡多与圆形清亮囊泡共存于同一轴—轴突触终末内。轴—轴突触均为对称型突触,有时与树突或胞体相连形成轴—轴—树突触或轴—轴—体突触。突触球多为以树突和棘为中心的中心树突型突触球。此外在SNG内还观察到嵴突触,并联突触等连接形式。SNG内突触的复杂性表明传入冲动在该核中可能经过扩散、汇聚和突触前抑制等多种复杂的整合过程调节内脏活动。     

SP,CGRP,ENK样阳性纤维终末在猫骶髓后连合核内的分布特征及超微结构

本实验用免疫组化电镜技术对骶髓后连合核中SP祥、CGRP样、L-ENK样阳性终末进行了观察,结果表明:SP样阳性终末主要含圆形清亮小泡,间有少量颗粒囊泡,主要与中、小树突形成不对称型轴-树突触(93%);还可见到不对称型轴-体突触(5%);也可见到少量的轴-轴突触(2%),SP样阳性终末为突触后成分。CGRP样阳性终末以含圆形清亮小泡为主,有的终末内混有颗粒囊泡。大多数终末(89%)与树突构成轴-树突触,但以远侧树突为主;也有少数(6%)的CGRP样阳性终末与胞体形成轴-体突触;还观察到由阴性终末与CGRP样阳性终末构成的轴-轴突触。L-ENK样阳性终末以含圆形清亮小泡为主,有时可见散在的颗粒囊泡,多与中、小树突形成不对型轴-树突触(92%);也观察到轴-体突触(5%)和轴-轴突触(3%)。     

猫后根初级传入纤维终未在后索核内的超微结构及其突触联系

用顺行溃变方法对猫后索核内后根初级传入纤维终末的超微结构和突触联系进行了研究.在切断C4～T1和 L4～S1脊神经后根3～4天后,电镜下发现后索核内有三种溃变终末,出现最多的是电子致密型溃变,此外,也观察到了少量的神经微丝型溃变和电子透明型溃变.溃变的初级传入终末多数较大,含有圆形突触小泡.溃变初级传入终末作为突触前成分主要与后索核内的树突形成轴-树突触,而轴-体突触和轴-轴突触较少.此外,还观察到溃变轴突终末参与形成突触复合体.     

大鼠中缝大核内5-羟色胺样P物质样和亮氨酸脑啡肽样突触结构的免疫电镜观察

用免疫电镜法在大鼠中缝大核内观察到:(1) 5-羟色胺(5-HT)样阳性轴突终末与阴性胞体、阳性和阴性树突以及阴性轴突终末,分别形成轴-体突触、轴-树突触和轴-轴突触;阴性轴突终末与阳性胞体和阳性树突分别形成轴-体和轴-树突触;(2) P物质样(SP样)和亮氨酸脑啡肽(L-Enk)样阳性轴突终末与阳性和阴性的胞体和树突,以及阴性轴突终末与阳性胞体和树突分别形成轴-体突触和轴-树突触,L-Enk样阳性轴突终末之间形成轴-轴突触;(3) 上述5-HT、SP和L-Enk样结构所形成的突触中,阴性轴突终末与阳性树突所形成的轴-树突触最多见;(4) 上述阳性轴突终末内主要含透明圆形小泡。免疫反应产物为电子密度高的物质,主要沉积于膜性细胞器的表面、透明圆形小泡和部分颗粒囊泡内和小泡膜上。     

大鼠脊髓胶状质内含P物质(SP)及脑啡肽(ENK)神经终末的超微结构

水文应用超微免疫组织化学技术探讨正常及切断后根大鼠胶状质(SG)内P物质(SP)及脑啡肽(ENK)轴突终末的分布及其突触联系。电镜观察发现:SP或ENK免疫反应产物主要分布于轴突终末的大颗粒囊泡的内面,小清亮囊泡及线粒体的外表面,在未应用秋水仙素的情况下,ENK样免疫反应性也可出现于树突及核周质,但电子密度较低。大量的含SP或ENK的终末,分别和未标记树突形成突触。但也偶然见到肽能神经终末与未标记轴突或胞体接触或形成突触联系。常遇到三或四联体,亦偶可见到以含SP轴突终来为中心的突触球。切断后根,手术侧既可见到变性轴突(SP阳性)与未标记轴突接触,又可见到ENK阳性轴突终末与溃变轴突接触,但为数不多。因此认为:通过SP阳性一级传入细纤维传递的感觉信息既可汇集于神经元的一个树突又可多向传递于若干个神经元。ENK在胶状质的抑制作用主要为突触后,但ENK对含SP的一级传入纤维的突触前抑制也不能排除。     

Opioid neurons and pain modulation: an ultrastructural analysis of enkephalin in cat superficial dorsal horn

To clarify the circuitry through which opioid compounds modulate spinal and trigeminal nociceptive transmission, we have examined the synaptic associations formed by leucine-enkephalin-containing (enkephalin) neurons in the superficial dorsal horn of the cat. As described previously, punctate enkephalin immunoreactivity is concentrated in the marginal layer (lamina I) and in both the outer and inner layers of the substantia gelatinosa (lamina IIo and IIi). In colchicine treated cats, enkephalin perikarya are most numerous in lamina I and at the border between laminae I and II. Ultrastructural analysis reveals that enkephalin cells receive a diverse afferent input. The majority of afferent inputs are presynaptic to the enkephalin dendrites; few axosomatic synapses are seen. Among these presynaptic axonal profiles are unlabeled axons which resemble primary afferent terminals, including the characteristic central axonal varicosity. Enkephalin dendrites are also postsynaptic to enkephalin immunoreactive axons. Two types of enkephalin axonal profiles appear in the superficial dorsal horn. Class I profiles are only found in lamina I. These are large profiles which form few synapses; those synapses made are axodendritic. Class II enkephalin axons are smaller and are distributed in both layers I and II. While Class II axons most commonly form axo-dendritic synapses, they also form axo-axonic synapses with flat vesicle-containing profiles; the latter are generally presynaptic to the enkephalin terminals. Serial analysis further revealed that both the enkephalin and the flat vesicle-containing profile synapse onto a common dendrite. Although enkephalin axons frequently lie adjacent to round vesicle-containing profiles, anatomical evidence that opioid axons form synapses with this type of ending was not found. An additional type of enkephalin vesicle containing-profile is found in layer IIi; its morphological features do not clearly distinguish its axonal or dendritic origin. These endings are typically postsynaptic to unlabelled central endings, and provide minimal presynaptic input to other elements in the neuropil. Like some class II axons, these labelled profiles contain vesicles which cluster at the membrane immediately adjacent to unlabelled central axons. These results indicate that spinal enkephalin neurons receive a variety of synaptic inputs. These include inputs which may derive from primary afferent axons. Enkephalin neurons, in turn, influence nociceptive transmission predominantly through postsynaptic mechanisms. Finally, while we did not observe enkephalin terminals presynaptic in an axoaxonic relationship, the possibility that enkephalin neurons modulate the excitability of fine fiber nociceptive and nonnociceptive afferents via "nonsynaptic interactions" is discussed.     

Immunohistochemical study of delta and mu opioid receptors on synaptic glomeruli with substance P-positive central terminals in chicken dorsal horn

In an attempt to clarify the mechanism underlying the regulation of the release of substance P (SP) from the central axon terminals of the synaptic glomeruli in lamina II of the dorsal horn, we examined the expression patterns of delta and mu opioid receptors (DOR and MOR) in relation to those of enkephalin (ENK) and SP in the synaptic glomeruli. DOR, MOR, ENK and SP immunoreactivities in lamina II of the dorsal horn in the chicken were examined by confocal laser scanning and electron microscopies. DOR immunoreactivity was localized in both SP-positive central terminals and peripheral elements, while MOR immunoreactivity was only localized in the peripheral elements of the synaptic glomeruli. Both of the peripheral DOR- and MOR-immunoreactive elements were shown to be vesicle-containing dendrites by electron microscopy. Dual immunohistochemistry indicated that DOR, MOR and ENK immunoreactivities were located in distinct peripheral elements. On the basis of present results, the possible roles of DOR and MOR in the regulation of the release of SP from the central axon terminals in the synaptic glomeruli are discussed.     

大颗粒小泡非突触部位的胞吐——论神经肽释放的另一种形式

本文在以前的工作基础上,进一步用电镜及免疫细胞化学方法,研究了大颗粒小泡非突触部位胞吐作用。实验结果表明,切除大鼠刚髭部皮肤1—24小时之后,术侧延髓后角浅层大颗粒小泡胞吐比对照侧明显增多(P<0.01),术后3—9天复又下降(近似对照动物),术后14—15天又急剧上升(P<0.01)。这些胞吐大部分出现于延髓后角浅层四种轴突终末的非突触部位,少最也发生于树突及轴突中。从术后第6天开始,术侧P物质明显减弱,而甲硫-脑腓肽略有增强。研究结果提示;1)后角浅层胞吐增多,P物质下降及脑腓肽增高,反映了中枢内不同神经元对去传入神经的功能调整作用;2)大颗粒小泡在非突触部位释放神经肽,弥散地作用于远距离的受体,可能起着神经调制物的作用。     

Anatomical distribution and ultrastructural organization of the gabaergic system in the rat spinal cord. An immunocytochemical study using anti-GABA antibodies

γ-Aminobutyric acid (GABA)-containing elements have been studied by light and electron microscopy in the rat spinal cord, using immunocytochemistry with anti-GABA antibodies. Light microscopy showed immunoreactive somata localized principally in laminae I–III, and occasionally in the deeper laminae of the dorsal horn and in the ventral horn. Small somata were also observed around the central canal. Punctate GABA-immunoreactive profiles were particularly concentrated in laminae I–III, and moderately abundant in the deeper laminae and in the ventral horn where they were observed surrounding the unlabelled motoneurons.

At the ultrastructural level, the punctate profiles corresponded to GABA-containing axonal varicosities or small dendrites. GABA-immunoreactive varicosities were presynaptic to labelled or unlabelled dendrites and cell bodies. Some unlabelled terminals presynaptic to unlabelled dendrites received symmetrical synaptic contacts from GABA-immunoreactive terminals.

These results confirm data obtained withl-glutamate decar☐ylase immunocytochemistry, and support the role of GABA in pre- and postsynaptic inhibition in the spinal cord, respectively via axoaxonal and axosomatic or axodendritic synapses.     

Reciprocal axo-axonal synapses between the common inhibitor and excitor motoneurons in crustacean limb muscles

Summary Nerve terminals of the common inhibitor motoneuron in a crab (Eriphia spinifrons) limb closer muscle and in a crayfish (Procambarus clarkii) limb accessory flexor muscle make neuromuscular synapses with the muscle membrane (postsynaptic inhibition) as well as axo-axonal synapses with the terminals of the excitatory axon (presynaptic inhibition). That transmission is from the inhibitor to the excitor terminals at these axo-axonal synapses is indicated by the occurrence on the inhibitor membrane of presynaptic dense bars denoting sites of transmitter release. Axo-axonal synapses with the opposite polarity, in which transmission is from an excitatory onto an inhibitory terminal, were occasionally seen either adjacent to or separate from the inhibitory axo-axonal synapse. Nerve terminals of the specific inhibitor in the crayfish opener muscle were seen to make numerous axo-axonal output synapses upon excitatory nerve terminals but excitor nerve terminals were not seen to make output synapses onto inhibitor terminals. Thus reciprocal axo-axonal synapses appear to be a feature of the common inhibitor but not of the specific inhibitor. The excitor-to-inhibitor component of these reciprocal synapses may serve to limit transmitter output in the common inhibitor axon by activating glutamate_B receptors which facilitate efflux of K⁺ and hyperpolarization of the membrane.     

Different kinds of axon terminals forming symmetric synapses with the cell bodies and initial axon segments of layer II/III pyramidal cells. II. Synaptic junctions

Summary Four different types of axon terminals form symmetric synapses with the cell bodies and initial axon segments of pyramidal cells in layer II/III of rat visual cortex. One type belongs to chandelier cells, and the other three kinds of terminals have origins that have not been established yet. These latter are referred to as large, medium-sized and dense terminals. The purpose of the present study was to examine the synaptic junctions formed by all four types of terminal. The synapses formed by the chandelier cell terminals are readily recognized in thin sections because of the characteristic features of both the terminals and the initial axon segments, which are the neuronal elements postsynaptic to them. In en face views of these axo-axonal synapses the junctions can be seen to have presynaptic dense projections that form a grid in which they are triagonally spaced, and have an average centre-to-centre spacing of 84 nm. As an ensemble the projections form the presynaptic grid, which usually has an oval or round outline, but may be notched on one side where projections are absent. The synaptic junctions of the large, medium-sized and dense terminals were examined by making reconstructions of the terminals from serial thin sections. It was found that at the interfaces between the axon terminals and the cell bodies of pyramidal cells, several separate synaptic junctions may be present, in addition to a number of puncta adhaerentia. Thus, there may be as many as five separate synaptic junctions and as few as one. It was also found that while the proportion of the area of the synaptic interface occupied by synaptic junctions was between 12% and 26% for dense terminals, for medium sized terminals it was 10–15%, and for the one large terminal reconstructed it was only 8%. Thus, there can be multiple synaptic junctions between each of these types of axon terminals and a pyramidal cell, and because many of the terminals forming symmetric junctions are boutons en passant, a number of vesicle release sites exist between the presynaptic axon and its postsynaptic partner. The axon terminals forming symmetric synapses in the cerebral cortex are assumed to be inhibitory, and consequently it is suggested that this arrangement of multiple release sites is designed to ensure that stimulation of the presynaptic axon results in an effective level of hyperpolarization of the postsynaptic neuron.     

家兔下橄榄背侧副核的超微结构特点——电镜研究

本实验对7只家兔的下橄揽背侧副核进行了电镜观察。除发现已报道过的轴树突触、轴体突触及以树突为中心的突触球外,还见此核内有轴轴突触、轴突与胞体棘形成的突触,以轴突为中心的突触球及核内微纤维束。轴树突触最常见,突触后成分为树突或棘。轴体突触较少见。有一轴突终末与一胞体棘形成突触,同时还与一树突形成突触。轴轴突触前、后成分中均含圆形囊泡,有时形成轴-轴-树突触串。家兔下橄榄背侧副核内见有两种突触球,一种以树突为中心,另一种以轴突为中心。还发现两个轴突终末同时与另一轴突终末形成轴→轴→轴突触。下橄榄背侧副核内的复杂突触形式,说明传入冲动在其中可能经过扩散、会聚、突触前抑制及整合等复杂的过程。     

三叉神经尾侧脊束核内P物质纤维终末的超微结构

目的　进一步探讨三叉神经尾侧脊束核内SP免疫反应阳性纤维在感觉传递中的可能作用。方法　SP采用免疫细胞化学方法和电子显微镜方法 ,观察大鼠三叉神经尾侧脊束核内SP阳性标记纤维的超微结构和突触联系。结果　SP轴突终末分布于树突间 ,这些轴突终末含有大量的透明小泡、少量大致密芯小泡和线粒体。经过秋水仙素处理后 ,可见到SP免疫反应阳性树突。多数SP轴突终末与非标记树突 ,以及个别SP轴突终末与SP树突形成轴 树突触。含SP的突触复合体较为多见 ,为会聚型。其中可见SP轴突终末与中心的非标记树突形成GrayⅡ型轴 树突触 ;另有非标记的轴突终末与中心SP树突形成 (扁平小泡形 )F型轴 树突触。结论　三叉神经尾侧脊束核接受多种纤维传入 ,SP纤维只是多种传入纤维中的一种。形态学证明 ,在痛调制活动中 ,三叉神经尾侧脊束核内有SP纤维构成的突触后抑制类型 (GrayⅡ )突触参与。