参环毛蚓神经组织的NSE、NF200、ED1、GFAP和NO细胞化学特性

为研究环节动物的神经组织学特性,我们选择生活在我国的环节动物门典型代表参环毛蚓（Pheretima aspergillum)为研究对象,使用若干种兔抗鼠抗体,进行了免疫细胞化学与细胞化学染色,在光学显微镜下观察反应阳性细胞的形态与分布。研究发现,在参环毛蚓脑,咽下神经节,腹神经节所有神经细胞呈NSE阴性;在参环毛蚓脑,部分神经细胞呈NF200阳性,在咽下神经节,腹神经节未观察到NF200阳性神经细胞;在参环毛蚓脑观察到较多ED1阳性细胞;在参环毛蚓脑,咽下神经节,腹神经节均未观察到GFAP阳性细胞;在参环毛蚓脑未观察到NADPH－d阳性神经细胞,而在咽下神经节和腹神经节部分神经细胞及纤维NADPH－d阳性。结果表明,参环毛蚓神经细胞的神经细胞特异的烯醇化酶特性,神经微丝蛋白特性与呈型胶质细胞的GFAP特性不同于哺乳动物;其神经组织存在有数量较多的吞噬功能的类似于哺乳动物小胶质细胞的细胞; 参环毛蚓的脑不含有NO能神经细胞,而咽下神经节和腹神经节含有NO能神经细胞。     

神经营养因子受体TrkA、TrkB、TrkC和TrkE在参环毛蚓体内的分布

高亲和性神经营养因子受体Trk,广泛存在分布于哺乳神经组织,研究表明：随着动物进化程度的降低,亚型Trk受体的类型有所减少,在低等动物,关于环节动物（Annelida)Trk受体存在与否,分布如何尚未见报道,以我国环节动物门典型代表参环毛蚓（Pheretima aspergillum)为研究对象,运用免疫细胞化学染色技术,在光镜下观察TrkA,TrkB,TrkC和TrkE阳性细胞和纤维的形态与分布,发现Trk存在于参环毛蚓的神经系统和非神经组织,各亚型分布有所差异,TrkA阳性 经细胞存在于咽下神经节和肠神经系,阳性神经纤维存在于腹神经索和肠神经系,TrkB阳性细胞存在于非神经组织的肠上皮,TrkC阳性神经细胞存在于脑,咽下神经节和肠神经细胞,阳性神经纤维存在于围咽神经环,腹神经索和肠神经系,TrkE阳性神经细胞存在于脑,阳性神经纤维存在于咽下神经节和腹神经素,上述研究结果表明：Trk存在于进化程度较的环节动物,Trk在进化上具有悠久的历史,各亚型Trk受体的不同分布提示不同部位的神经细胞受不同神经营养因子的作用。     

十三种神经递质和神经肽在参环毛蚓神经系统的分布:免疫细胞化学研究

选择生活在我国的环节动物门典型代表参环毛蚓（Pheretima aspergillum)的研究对象,使用13种抗体,运用免疫细胞化学染色技术,在光学显微镜下观察神经递质和神经肽阳性细胞的形态与分布。研究发现在参环毛蚓脑,ACTH阳性神经细胞、AVP阳性神经细胞、calcitonin阳性神经细胞、CCK阳性神经细胞、glutamate阳性神经细胞和NPY阳性神经细胞染色浓重,分布密集,位于脑的后侧半,尤其特别的是在calcitonin阳性大神经细胞内含有粗大密集的分泌颗粒。β－EP阳性神经细胞染色较谈,OT阳性神经细胞染色也较深,但分布稀疏, calbindi-D、CGRP、GABA、SOM、VIP呈阴性反应;在咽下神经节与腹神经节,仅有少量AVP阳性神经细胞、glutamate阳性神经细胞和NPY阳性神经细胞散在分布;在肠神经系统,β－EP阳性神经细胞、CGRP阳性神经细胞、SOM阳性神经细胞、VIP阳性神经细胞染色较深,分布散在;另外,在肠上皮,CGRP阳性上皮细胞、calcitonin上皮细胞和VIP阳性上皮细胞呈强阳性反应,分布于阴性上皮细胞之间。表明表明参环毛蚓神经系统存在着在其它环节动物所观察到的神经递质和神经肽：ACTH、VIP、β-endorphin、CCK、NPY、CGRP,亦存在着文献从未报道过的神经递质和神经肽：calcitonin、AVP、OT、glutamate、SOM,其中calcitonin、CGRP、VIP双重分布于神经细胞和非神经组织的肠上皮细胞,本文首次提供了形态学的证据。     

柞蚕幼虫期神经系统一氧化氮合酶的NADPH-d组织化学

运用NADPH-d组织化学整体染色方法研究柞蚕Antheraea pernyi Gu rin-Meneville幼虫神经系统中一氧化氮合酶阳性细胞的分布、数量及形态特征。结果表明,柞蚕各龄期幼虫中枢神经的脑及各神经节中都有一氧化氮合酶阳性反应,阳性神经元根据其形态大小和染色特性可分为A,B,C3种类型:A型细胞沿神经节中线分布,阳性反应较强,胞体长径约30~70μm,各神经节中的数量恒定。B型细胞多分布于神经节周边部分,阳性反应较弱,胞体长径约8~20μm,各神经节中的数量变化较大,随着龄期增加有减少的趋势。C型细胞分布于咽下神经节和第8腹神经节,在3种细胞中阳性反应最强,胞体长径约16~50μm。     

两种软体动物神经系统一氧化氮合酶的组织化学定位

运用一氧化氮合酶(NOS)组织化学方法研究了软体动物门双壳纲种类中国蛤蜊和腹足纲种类嫁Qi神经系统中NOS阳性细胞以及阳性纤维的分布。结果表明：在蛤蜊脑神经节腹内侧,每侧约有10-15个细胞呈强NOS阳性反应,其突起也呈强阳性反应,并经脑足神经节进入足神经节的中央纤维网中;足神经节内只有2个细胞呈弱阳性反应,其突起较短,进入足神经节中央纤维网中,但足神经节中,来自脑神经节阳性细胞和外周神经系统的纤维大多呈NOS阳性反应;脏神经节的前内侧部和后外侧部各有一个阳性细胞团,其突起分别进入后闭壳肌水管后外套膜神经和脑脏神经索。脏神经节背侧小细胞层以及联系两侧小细胞层的纤维也呈NOS阳性反应。嫁Qi中枢神经系统各神经节中没有发现NOS阳性胞体存在;脑神经节、足神经节、侧神经节以及脑—侧、脑—足、侧—脏连索中均有反应程度不同的NOS阳性纤维,这些纤维均源于外周神经。与已研究的软体动物比较,嫁Qi和前鳃亚纲其它种类一样,神经系统中NO作为信息分子可能主要存在于感觉神经。而中国蛤蜊的神经系统中一氧化氮作为信息分子则可能参与更广泛的神经调节过程。     

封面故事

<正>生物体内存在着复杂的一氧化氮合酶(nitric oxide synthase,NOS)活性调节机制以精确调控一氧化氮的生成。在神经系统中,一氧化氮主要由神经型一氧化氮合酶(neuronal nitric oxide synthase,nNOS)催化生成     

中华蜜蜂咽下神经节的结构和胚后发育

咽下神经节是昆虫腹神经索的第一个复合神经节, 主要调节口器附肢的活动。本研究通过形态解剖、 BrdU免疫组织化学等技术, 对中华蜜蜂Apis cerana cerana咽下神经节的组织结构和胚后发育过程进行了比较研究。结果表明： 中华蜜蜂的咽下神经节由上颚、 下颚和下唇3个神经节组成。在胚后发育过程中, 细胞增殖的活跃期主要集中在预蛹和蛹发育的第1天, 增殖活动一直持续到蛹发育的第4天结束。根据神经胶质细胞的位置和形态, 咽下神经节中的神经胶质可分为3种类型--表面神经胶质、 皮层神经胶质和神经纤维网神经胶质。本研究为蜜蜂神经系统的发育和功能研究提供了理论基础。     

神经型一氧化氮合酶的活性调控

一氧化氮是重要的信使分子,在生物体内参与众多生理及病理过程。生物体内存在着复杂的一氧化氮合酶活性调控机制以精确调控一氧化氮的生成。在神经系统中,一氧化氮主要由神经型一氧化氮合酶催化生成。神经型一氧化氮合酶的活性主要受到翻译后水平上钙离子和钙调蛋白的调控,其调控方式包括二聚化、多位点的磷酸化和去磷酸化,以及主要由PDZ结构域介导的蛋白质-蛋白质相互作用。一氧化氮本身对其合酶的活性具有负反馈调控作用。近年来的研究提示,细胞质膜上的脂筏微区在神经性一氧化氮合酶的活性调控中也起到重要的调节作用。     

地龙类药用动物的比较鉴别

调查发现我国中药地龙类药用动物主要有14个种,分别隶属于钜蚓科、正蚓科和链胃蚓科4个属。包括参环毛蚓Phereti- ma aspergillum(E Perrier)、通俗环毛蚓Pheretima vulgaris(Chen)、威廉环毛蚓Pheretima guillelmi(Michaelsen)、栉盲环毛蚓Phere- tima pectinifera(Michaelsen)、湖北环毛蚓Pheretima hupeiensis(Michaelsen)、直隶环毛蚓Pheretima tshiliensis(Michaelsen)、白颈环毛蚓Pherctima califomica(Kinberg)、中材环毛蚓Pheretima medioca(Chen et Hsu)、秉前环毛蚓Pheretima pracpinguis(Gates)、秉氏环毛蚓Pheretima carnosa(Goto et Hatai)、异毛环毛蚓Pheretima diffringens(Baird)、赤子爱胜蚓Eisenia foetida(Savigny),背暗异唇蚓Allolbophora caliginosa subsp.trapezoides(Ant.Duges)以及日本杜拉蚓Drawida japonica(Michaelsen)。本文对我国地龙类原动物主要种类进行了分类及形态描述,并编制了检索表和检索图,以供鉴别,为地龙的开发利用提供资料。     

东亚飞蝗神经系统解剖的关键

获得完整的神经系统是蝗虫解剖实验的难点,使用福尔马林液浸标本解剖时神经易断裂,使用活体或经过冷冻、冷藏处理的东亚飞蝗为解剖材料,神经能保持弹性,在分离咽下神经节和围咽神经环时小心去除头壳内幕骨,可获得完整的神经系统。     

Distribution of NADPH-diaphorase and nitric oxide synthase in the trigeminal ganglion and mesencephalic trigeminal nucleus of the cat. A histochemical and immunohistochemical study

The trigeminal ganglion (TrG) and mesencephalic trigeminal nucleus (MTN) neurons are involved in the transmission of orofacial sensory information. The presence of nitric oxide (NO), a putative neurotransmitter substance in the nervous system, was examined in the cat TrG and MTN using nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry and nitric oxide synthase (NOS) immunohistochemistry. In the TrG, where the majority of the trigeminal primary afferent perikarya are located, most of the intensely NADPH-d/ NOS-stained cells were small in size and distributed randomly throughout the ganglion. The medium-sized neurons were moderately stained. A plexus of pericellular varicose arborizations around large unstained ganglion cells and densely stained fibers in-between could also be observed. In the caudal part of the MTN, both NADPH-d activity and NOS immunoreactivity was present in MTN neurons. In addition, a few scattered NADPH-d/NOS-containing neurons were found in the mesencephalic-pontine junction part of the nucleus. In contrast, only nerve fibers and their terminals were present at a more rostral level in the mid- and rostral MTN. MTN neuronal perikarya were enveloped in fine basket-like NADPH-d/ NOS-positive networks. Differential expression patterns of NOS and its marker NADPH-d suggest that trigeminal sensory information processing in the cat MTN is controlled by nitrergic input through different mechanisms. We introduce the concept that NO can act as a neurotransmitter in mediating nociceptive and proprioceptive information from periodontal mechanoreceptors but may also participate in modulating the activity of jaw-closing muscle afferent MTN neurons.     

Expression and colocalization of NADPH-diaphorase and heme oxygenase-2 in trigeminal ganglion and mesencephalic trigeminal nucleus of the rat

Carbon monoxide (CO) and nitric oxide (NO) are two endogenously produced gases that can function as second messenger molecules in the nervous system. The enzyme systems responsible for CO and NO biosynthesis are heme oxygenase (HO) and nitric oxide synthase (NOS), respectively. The present study was undertaken to examine the distribution of HO-2 and NOS of the trigeminal primary afferent neurons of the rat, located in the trigeminal ganglion (TG) and mesencephalic trigeminal nucleus (MTN), using histochemistry and immunohistochemistry. NADPH-d staining was found in most neurons in TG. The intensely NADPH-d-stained neurons were small- or medium-sized, while the large-sized neurons were less intensely stained. Immunocytochemistry for HO-2 revealed that almost all neurons in TG expressed HO-2, but they did not appear cell size-specific pattern. NADPH-d and HO-2 positive neurons appeared the same pattern, which was NADPH-d activity and HO-2 expression progressively declined from the caudal to rostral part of the MTN. A double staining revealed that the colocalization of NADPH-d/HO-2 neurons was 97.3% in TG and 97.6% in MTN. The remarkable parallels between NADPH-d and HO-2 suggest that NO and CO are likely neurotransmitters and mediate the orofacial nociception and sensory feedback of the masticatory reflex arc together.     

A large proportion of pelvic neurons innervating the corpora cavernosa of the rat penis exhibit NADPH-diaphorase activity

Nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry, which indicates the presence of neural nitric oxide synthase, the enzyme responsible for the generation of nitric oxide, was used in combination with retrograde labelling methods to determine, in whole-mounts and sections of rat major pelvic ganglia, whether neurons destined for the penile corpora cavernosa were able to produce nitric oxide. In whole-mount preparations of pelvic ganglia, among the 607±106 retrogradely labelled neurons innervating the penile corpora cavernosa, 84±7% were NADPH-diaphorase-positive, 30±7% of which were intensely histochemically stained. In serial sections of pelvic ganglia, out of a mean count of 451 retrogradely labelled neurons, 65% stained positively for NADPH-diaphorase. An average of 1879±363 NADPH-diaphorase positive cell bodies was counted in the pelvic ganglion. In the major pelvic ganglion, neurons both fluorescent for Fluorogold or Fast Blue and intensely stained for NADPH-diaphorase were consistently observed in the dorso-caudal part of the ganglia in the area close to the exit of the cavernous nerve and within this nerve. This co-existence was much less constant in other parts of the ganglion. In the rat penis, many NADPH-diaphorase-positive fibres and varicose terminals were observed surrounding the penile arteries and running within the wall of the cavernous spaces. This distribution of NADPH-diaphorase-positive nerve cells and terminals is consistent with the idea that the relaxation of the smooth muscles of the corpora cavernosa and the dilation of the penile arterial bed mediated by postganglionic parasympathetic neurons is attributable to the release of nitric oxide and that nitric oxide plays a crucial role in penile erection. Moreover, the existence in the pelvic ganglion of a large number of NADPH-diaphorase-positive neurons that are not destined for the corpora cavernosa suggests that nitric oxide is probably also involved in the function of other pelvic tissues.     

Nitric oxide synthase-containing nerve fibers and neurons in the genital tract of the female mouse

Nitric oxide (NO) is generated intracellularly from L-arginine by the action of the enzyme nitric oxide synthase (NOS). The present investigation demonstrates immunoreactivity against NOS and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase activity in nerve cells and fibers of the reproductive system of the female mouse. The density of nerve fibers staining for NOS varied among different genital organs. The ovary and Fallopian tube were devoid of NOS-positive nerves. The uterine horns received sparse innervation by NOS-containing nerve fibers. The most abundant NOergic innervation was found in the uterine cervix and vagina, where the nerve fibers ran parallel to the smooth muscle bundles and beneath the epithelium; they also accompanied intramural blood vessels. The vaginal muscular wall contained single or groups of NOS-reactive nerve cells. Clusters of NOS-containing neurons were located in Frankenhäuser's ganglion at the cervico-vaginal junction. NO may therefore act as a transmitter in the nervous control of the female reproductive tract.     

Localization of nitric oxide synthase in rat trigeminal primary afferent neurons using NADPH-diaphorase histochemistry

Summary Nitric oxide (NO) is a ubiquitous gaseous neurotransmitter that has been ascribed to a large number of physiological roles in sensory neurons. It is produced by the enzyme nitric oxide synthase (NOS). To identify the NOS-containing structures of rat trigeminal primary afferent neurons, located in the trigeminal ganglion (TrG) and mesencephalic trigeminal nucleus (MTN), histochemistry to its selective marker nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) was applied in this study. In the TrG approximately half of the neuronal population was NADPH-d reactive. Strongly positive were neurons mainly of small-to-medium size. Neuronal profiles of large diameter were less intensely stained. In addition, NADPH-d-positive nerve fibers were dispersed throughout the ganglion. Nitrergic neurons were located in the caudal part and mesencephalic-pontine junction of the MTN. Most of them were large-sized pseudounipolar cells. In a more rostral aspect, the reactive psedounipolar MTN profiles gradually decreased in number and intensity of staining. There, only a fine meshwork of stained thin fibers and perisomatic terminal arborizations, and also some isolated perikarya of NADPH-d stained multipolar MTN neurons, were observed. The predominant NADPH-d localization in smaller in size TrG neurons, which are considered nociceptive, suggests that NO may play a role in the pain transmission in the rat trigeminal afferent pathways. In addition, the wide distribution of NADPH-d activity in large pseudounipolar and certain multipolar MTN neurons provides substantial evidence that NO may also participate in mediating proprioceptive information from the orofacial region. The differential expression patterns of nitrergic fibers in the TrG and MTN suggest that trigeminal sensory information processing is controlled by nitrergic input through different mechanisms.     

豚鼠小肠神经节丛的NADPH—黄递酶组织化学观察

目前已知,ＮＡＤＰＨ－－黄递酶组化法可选择性地显示－－氧化氮合成酶（ＮＯｓｙｎｔｈａｓｅ,ＮＯＳ）神经元。因此,我们以ＮＡＤＰＨ－黄递酶组化法,观察了豚鼠小肠肌间神经丛和粘膜下神经丛的神经网格以及ＮＯＳ神经元。结果表明,三段小肠肌间神经丛的神经网眼大小和形态有明显差异,与对应的粘膜下神经丛相比,差异更显著。在肌间神经丛中,ＮＡＤＰＨ－黄递酶阳性神经元胞体大小不等;其长突起伸入节间束,而短突起较多,并可见短突起彼此连接．构成节内偶见的局部神经元回路。从小肠上段到下段,ＮＯＳ神经元数量呈下降趋势。在粘膜下神经丛,我们也观察到少数ＮＯＳ神经元。     

Distribution and morphological features of nitrergic neurons in the porcine large intestine

The distribution of nitric oxide synthase (NOS), an enzyme involved in the synthesis of the presumed non-adrenergic noncholinergic inhibitory neurotransmitter nitric oxide (NO), was demonstrated in the enteric nervous system of the porcine caecum, colon and rectum. Techniques used were NOS-immunocytochemistry and nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd)-histochemistry. Throughout the entire large intestine, NOS-immunoreactive (IR) and NADPHd-positive neurons were abundant in the myenteric and outer submucous plexus. In the inner submucous plexus, only a small number of positive neurons were found in the caecum and colon, while a moderate number was observed in the rectum. The nitrergic neurons in the porcine enteric nerve plexuses were of a range of sizes and shapes, with a small proportion showing immunostaining for vasoactive intestinal polypeptide. Varicose and non-varicose NOS-IR and NADPHd-positive nerve fibres were present in the ganglia and connecting strands of all three plexuses. Nerve fibres were also numerous in the circular muscle layer, scarce in the longitudinal muscle coat and negligible in the mucosal region. The abundance of NOS/NADPHd in the intrinsic innervation of the caecum, colon and rectum of the pig implicates NO as an important neuronal messenger in these regions of the gastrointestinal tract.     

人胎胃壁内NOS阳性神经元发育的研究

研究用NADPH-d组织化学法对第3个月胎龄至足月人胎胃壁内NOS阳性神经元的分化,迁移和生长发育进行了观察。结果表明：第5个月龄时,肌层神经节处的圆形细胞出现一氧化氮合酶（nitric oxide synthase,NOS)阳性反应。第6个月龄时,NOS阳性细胞分化演变成NOS阳性神经细胞,细胞呈圆形或椭圆形,核大,细胞质极少,由细胞发出短小的6个月龄时,NOS阳性细胞分化演变成NOS阳性神经细胞,细胞呈圆形或椭圆形,核大,细胞质极少,由细胞发出短小的突起,有部分NOS阳性细胞分化演变成梭形的NOS阳性神经细胞,呈条索状排列和粘膜下层延伸,吸的到达粘膜层,在粘膜层形成网状细胞,第7个月龄时,神经元细胞明显增大,细胞质增多,染色加深,在肌层形成神经节,神经节细胞突起投射到整个肌层,第8-10个月龄时,肌层和粘膜下层神经元日趋成熟,细胞质增多,染色强度加深,肌层神经纤维分布密度增加,大多数神经纤维增粗,有的呈弹簧样弯曲,其走向与肌纤维长轴平行。结果提示;人胎胃壁内NOS阳性神经元来源于胚胎早期肌层神经节处的圆形细胞,通过分化,生长发育形成成熟的NOS阳性神经元。