雌性大鼠生殖器官一氧化氮合酶1免疫反应神经纤维的分布

实验采用亲和免疫组化ＳＡＢＣ法对雌性大鼠动情间一氧化氮合酶的１正常分布进行了观察。一氧化氮合酶１免疫反应阳性神经纤维在生殖管道多数呈细丝状弯曲走行，曲线体处深染不典型，分布以宫颈、宫体和阴道较多，宫角量中等，在卵巢未发现明显的纤维，在卵巢未发现明显的纤维，在各生殖管道，神经纤维主要与肌束同方向直地或包绕在血管周。分析雌性大鼠生殖器官内一氧化氮合酶１免疫反应阳性神经纤维的分布，提示一氧化氮可能与血管     

P物质免疫反应阳性神经纤维在雌性大鼠生殖器官的分布

目的和方法:实验采用亲和免疫组化SABC法对雌性大鼠动情间期P物质的正常分布进行了观察.结果:P物质免疫反应阳性纤维在子宫角与阴道分布最为丰富,宫颈和宫体分布中等,卵巢内免疫阳性神经纤维较少,且免疫阳性神经纤维在子宫角有从卵巢端向阴道端递减的趋势,在各管状生殖道,免疫阳性神经纤维以内膜和外膜分布较多.结论:分析研究雌性大鼠生殖器官内P物质免疫反应阳性神经纤维的分布,提示P物质可能与卵泡发育、性激素的产生、血管及非血管平滑肌的运动调节有关.     

大鼠心肌氮能神经元及纤维的分布

目的　观测大鼠心脏各部心肌一氧化氮合酶 (nNOS)免疫阳性神经元的形态及分布 ,为探讨氮能神经在心肌的作用提供形态学资料。方法　采用免疫组织化学SABC法显示大鼠心脏nNOS免疫阳性神经元及神经纤维的形态与分布。结果　在大鼠各部心肌内均有氮能神经元和神经纤维分布 ,胞体呈多种形状。nNOS免疫阳性神经纤维呈串珠状、条索状、细线状及不定形状 ,其走行多与肌纤维长轴平行。氮能神经元及纤维的数密度和面密度心房高于心室 ,6月龄高于 1月龄组 (P <0 .0 1)。结论　各部心肌均有氮能神经元及神经纤维 ,心房的含量高于心室。     

豚鼠脑底动脉神经肽Y免疫反应阳性神经纤维的分布

本文在脑底动脉整体剥离标本上,用ABC免疫组织化学法结合葡萄糖氧化酶-DAB-硫酸镍铵(Glucose oxidase—DAB—Nickel,GDN)显色技术,对豚鼠脑底动脉神经肽Y(Neuropeptide Y,NPY)免疫反应阳性神经纤维分布进行了研究。结果发现,含NPY免疫反应阳性神经纤维广泛分布于脑底动脉及其分支,在脑底动脉吻侧的NPY阳性神经纤维的密度明显高于尾侧。在动脉分支的起始部纤维密度更高。NPY阳性神经纤维主要走行在血管外膜中,多数呈膨体型,少数为非膨体型。在脑底动脉吻侧血管,纤维呈密集的网格状,而在尾侧呈螺旋状走行,在基底动脉和椎动脉则主要为纵向走行,上述结果提示,脑底动脉各段NPY免疫反应阳性神经纤维的分布密度和形态差异可能与其发生及脑血流的生理调节有关。     

一氧化氮合酶在大鼠附睾内的表达及其可能的作用

目的　为了阐明一氧化氮合酶 (NOS)在附睾内的表达 ,同时探讨NO在男性生殖功能中的作用。方法　本研究采用正常大鼠附睾组织 ,用免疫组织化学ABC法研究两种NOS(iNOS、eNOS)的表达。结果　在附睾上皮细胞内eNOS及iNOS均为免疫反应阳性 ,其中eNOS表达强 ,iNOS表达较强 ,血管内皮及平滑肌细胞内仅为eNOS免疫反应阳性 ,附睾间质中仅iNOS为免疫反应阳性。结论　两种NOS在附睾内均有不同程度的表达 ,各有特定的分布区域 ,提示NOS与附睾的功能密切相关     

大鼠肺一氧化氮合酶阳性神经结构

以还原型尼克腕腺嘌呤二核苷酸磷酸辛酰胺脱氢酶组织化学技术研究大鼠肺一氧化氮合酶阳性神经结构。结果表明：（１）：气管，支气管树系统壁内神经节部分神经细胞为一氧化氮合酶阳性神经细胞；（２）部分强阳性神经细胞的突起穿越神经节延向管壁粘膜组织或肺组织；（３）在各级肺动，静脉及肺组织内分布有不同密度的一氧化氮合酶阳性神经纤维。     

大鼠脑内血管的胆硷能神经纤维的分布

本文应用ＡＢＣ免疫过氧化物酶法，以胆硷乙酰转移酶（ＣｈＡＴ）作为标记物，对１２只Ｗｉｓｔａｒ大鼠脑实质内血管的免疫阳性神经纤维的分布类型，纤维密度进行了观察。结果是：端脑皮质区（第Ⅰ感觉运动皮质，第Ⅱ感觉运动皮质，中央前区，纹状皮质）的血管；海马（ＣＡ１区，ＣＡ２区）血管；下丘脑（前区、外侧区）血管；脑桥（被盖部）及延髓实质内血管均可见明显免疫反应阳性胆硷能神经纤维分布。纤维呈棕褐色，多数为单一细线状结构。纤维走行不一，有的与血管长轴垂直走行或斜行，有的与血管长轴平行。可见类似膨体样结构。各部脑实质内血管阳性纤维分布均较稀疏。     

神经肤Y免疫反应阳性神经纤维在大鼠脾脏内的分布

用ABC免疫组织化学法结合葡萄糖氧化酶-DAB-硫酸镍铵(GDN)显色技术,研究神经肽Y(NPY)免疫反应阳性神经纤维在大鼠脾脏内的分布。结果表明,在脾脏内有较丰富的NPY免疫反应(NPY-IR)阳性神经纤维,它们多呈串珠状,主要伴动脉及其分支走行,也见于脾被膜的结缔组织内,白髓、红髓和边缘区等部位的淋巴组织中,以及脾血窦周围。脾内NPY免疫反应阳性神经纤维与血管和淋巴细胞的关系密切,提示它们对脾淋巴细胞的发育和功能可能有调节作用。NPY可能直接作用于淋巴细胞或通过调节脾的血液循环间接地发挥作用。     

生殖周期中小鼠子宫NOS定位及血清NO水平变化

目的　系统研究 3种一氧化氮合酶 (nitricoxidesynthase ,NOS)在生殖周期中小鼠子宫分布变化 ,探讨内源性一氧化氮 (nitricoxide,NO)在生殖周期中可能的生理作用。　方法　免疫组织化学LSAB法 ,比色法。　结果　神经型一氧化氮合酶 (nNOS ,NOS1)在生殖周期小鼠子宫有较为稳定的表达 ,大量的阳性细胞主要分布在子宫内膜上皮 ,内膜基质 ,血管内皮 ,腺上皮及肌层 ;内皮型一氧化氮合酶 (eNOS ,NOS3)在妊娠中期小鼠子宫表达明显增强 ,蜕膜上皮 ,腺上皮和血管内皮呈强阳性标记 ;诱导型一氧化氮合酶 (iNOS ,NOS2 )在妊娠早期表达最强 ,妊娠中、晚期表达稍下降。阳性标记主要分布在肌层 ,血管内皮 ,腺上皮 ,内膜上皮及内膜基质 ;然而动情期小鼠子宫未见iNOS阳性标记。此外 ,还测定了生殖周期小鼠血清NO水平变化。　结论　 3种NOS同工酶在生殖周期小鼠子宫可能既协作又分工 ,由其产生的内源性NO对雌鼠动情、胚胎植入、分娩前子宫静息状态的维持、分娩始动以及产后子宫修复等生理过程可能均有调节作用。     

大鼠第三脑室一氧化氮合酶阳性触液神经元的分布及与催产素的共存

应用 NADPH-d组织化学方法观察了大鼠第三脑室一氧化氮合酶阳性触液神经元的分布及其形态 ,并结合免疫组织化学技术研究了一氧化氮合酶与催产素在第三脑室触液神经元内的共存。结果显示 :在视前区至室间核后大细胞亚核平面的第三脑室室壁均有一氧化氮合酶阳性触液神经元分布 ,呈自前向后 ,由腹侧部渐向背侧部过渡迁移的特征。一氧化氮合酶阳性触液神经元绝大部分为大细胞型标记神经元 ,胞体呈卵圆形、梭形、多角形与倒置梨形。它们位于脑室管膜内、室管膜下 ,或距室管膜有一定距离处 ,但有突起伸至第三脑室。第三脑室催产素免疫阳性触液神经元的形态及分布与一氧化氮合酶阳性触液神经元基本相似 ,两者高度共存。催产素 /一氧化氮合酶双标神经元 ,约占阳性细胞总数的 90 .9%。上述三种阳性触液神经元与邻近核团关系密切 ,尤其与室旁核之间有很多的阳性纤维互相交错。本研究结果表明 ,第三脑室有大量的一氧化氮合酶与催产素免疫阳性神经元分布 ,并且高度共存 ,从而建立了催产素的下丘脑 -脑脊液 -垂体神经体液调节环路的结构基础 ,对生殖与性行为起着重要的调控作用     

No colocalization of immunoreactivities for VIP and neuronal NOS, and a differential relation to cGMP-immunoreactivity in bovine penile smooth muscle

The distribution of immunoreactivity (IR) for the neuropeptide vasoactive intestinal polypeptide (VIP) and neuronal nitric oxide synthase (nNOS) in the bovine retractor penis muscle (RP) and penile artery (PA) was studied by using two different methods. The distribution of these immunoreactivities was also compared with that of the immunoreactivity for cyclic guanosine monophosphate (cGMP). In both tissues the nerve fibers and terminals immunoreactive for VIP had a distribution that was completely different from that of the nerve fibers and terminals immunoreactive for nNOS. This contrasts with the previous observations in penile smooth muscle of other species. In the RP, as well as in the PA, many of the VIP-IR fibers were also immunoreactive for neurofilaments (NF), whereas the nNOS-IR fibers were consistently devoid of NF-IR. Stimulation with sodium nitroprusside, a nitric oxide donor, considerably increased cGMP-IR in the smooth muscle cells in both RP and PA, and in several nerve fibers in PA. Many of these cGMP-IR nerve fibers exhibited nNOS-IR, whereas none of them was immunoreactive for VIP. Our results suggest that the degree of coexistence of VIP-IR and nNOS-IR in the nerve fibers and terminals innervating penile smooth muscle show wide species differences. They also suggest that the mechanisms by which VIP could be involved in neurogenic penile erection may vary between species.     

Distribution of NADPH-diaphorase-positive nerves in the uterine cervix and neurons in dorsal root and paracervical ganglia of the female rat.

Nitric oxide synthase (NOS) was selectively stained in nerve fibers of the uterine cervix and neurons of the paracervical (PG) and dorsal root ganglia (DRG) by NADPH diaphorase histochemistry. In the cervix, numerous NADPH-diaphorase-positive nerve fibers were observed in the myometrium, endometrium and around arteries. In addition, a subpopulation of neurons within ganglia that innervate the cervix, i.e., the PG and DRG, were NADPH-diaphorase positive; thus the fibers in the cervix could be sensory and/or autonomic. NADPH-diaphorase/NOS localization identifies sites where nitric oxide (NO) can be synthesized. Since NO relaxes vascular and nonvascular smooth muscle, the prevalence and anatomical localization of NADPH-diaphorase-positive fibers suggest that they could influence functions of the uterine cervix.     

Gamma‐aminobutyric acid B Receptor Immunoreactivity in the Mouse Adrenal Medulla

The present study examined gamma‐aminobutyric acid B (GABA_B) receptor, GABA, choline acetyltransferase (ChAT), and neuronal nitric oxide synthase (nNOS) immunoreactivities in the mouse adrenal medulla. GABA_B receptor immunoreactivity was seen in numerous chromaffin cells and in a few ganglion cells of the adrenal medulla. By using a formaldehyde‐induced fluorescence (FIF) method, GABA_B receptor immunoreactivity was observed in numerous adrenaline (A) cells, but not in noradrenaline (NA) cells showing blue‐white fluorescence. This suggests that GABA_B receptors may be present in the A cells and be related to the secretory activity of A cells but not NA cells in the mouse adrenal medulla. GABA_B receptor immunoreactive ganglion cells were shown to be nNOS immunopositive by using a double immunostaining method. Weak GABA immunoreactivity was visible in some chromaffin cells and in the numerous nerve fibers of the medulla. By using the FIF method, weak GABA‐immunoreactive chromaffin cells were shown to be in the NA cells showing blue‐white fluorescence. GABA‐immunoreactive nerve fibers were in dense contact in A cells, but not NA cells. GABA‐immunoreactive nerve fibers closely contacted a few ganglion cells. Numerous GABA‐immunoreactive nerve fibers in the medulla showed ChAT immunoreactive. This result suggests that GABA and acetylcholine may be released from the same nerve fibers and may have a secretory effect on the A cells of the medulla. Anat Rec, 296:971–978, 2013. © 2013 Wiley Periodicals, Inc.     

Perivascular nerve fibres and endothelial cells of the rat basilar artery: immuno-gold labelling of antigenic sites for type I and type III nitric oxide synthase

Ultrastructural localisation of type I (neuronal) and type III (endothelial) isoforms of nitric oxide synthase in perivascular nerve fibres (axons) and endothelial cells was studied in the Wistar rat cerebral basilar artery, using monoclonal antibodies either to type I or type III nitric oxide synthase and post-embedding colloidal-gold immunocytochemistry. Labelling signal (gold particles) for type I and type III nitric oxide synthase was localised both in axons and endothelial cells. In the axon profiles, labelling for either type I or type III nitric oxide synthase was localised in the axoplasm and the lumen and/or membrane of small agranular synaptic vesicles. In the endothelial cells, labelling for either type-I or type-III nitric oxide synthase was predominantly in the cytoplasm. The present qualitative data extends our previous study of cerebrovascular nerve fibres and endothelial cells employing monoclonal antibodies; the localisation of nitric oxide synthase in a subpopulation of synaptic vesicles in nitric oxide synthase-positive cerebrovascular nerves suggests that vesicular mechanisms may be involved in the production/release of nitric oxide. © 1998 Chapman and Hall     

Projections and chemical coding of neurons with immunoreactivity for nitric oxide synthase in the guinea-pig small intestine.

The distribution of nitric oxide synthase (NOS) immunoreactivity was investigated in the guinea-pig small intestine. There were many immunoreactive nerve cell bodies in the myenteric plexus but very few in submucous ganglia. NOS immunoreactivity was not found in non-neuronal cells except for rare mucosal endocrine cells. Abundant immunoreactive nerve fibres in both myenteric and submucous ganglia, and in the circular muscle, arose from myenteric nerve cells whose axons projected anally along the intestine. NOS immunoreactivity coexisted with VIP-immunoreactivity, but not with substance P immunoreactivity. We conclude that nitric oxide synthase is located in a sub-population of enteric neurons, amongst which are inhibitory motor neurons that supply the circular muscle layer.     

Chemical coding of intrinsic and extrinsic nerves in the guinea pig gallbladder: distributions of PACAP and orphanin FQ

The complexity of the neural regulation of the gallbladder is reflected by the variety of neuroactive compounds that are found in the intrinsic and extrinsic nerves of the guinea pig gallbladder. The studies reported here used antisera to test for the presence of gallbladder nerves that are immunoreactive for the neuroactive peptides, pituitary adenylyl activating polypeptide (PACAP), and/or orphanin FQ (OFQ, also known as nociceptin). PACAP immunoreactivity was observed in nerve fibers of the paravascular plexus that were also immunoreactive for calcitonin gene-related peptide. These nerve fibers, which are also immunoreactive for substance P, could be followed into the ganglionated plexus. Within the ganglia, a small proportion of neurons was found to be immunoreactive for PACAP; these neurons were also immunoreactive for vasoactive intestinal peptide and nitric oxide synthase. Immunoreactivity for OFQ was observed in the perivascular plexus in nerve fibers that were also immunoreactive for tyrosine hydroxylase. These nerves were previously shown to be immunoreactive for neuropeptide Y. In the ganglionated plexus, immunoreactivity was observed in all gallbladder neurons, as demonstrated by double staining with antiserum directed against the neuron-specific RNA binding protein, Hu. OFQ immunoreactivity was also present in the small catecholaminergic neurons that are observed in a subset of the ganglia. These results further demonstrate the neurotransmitter diversity of the nerves of the gallbladder, and they provide an incentive for studies of the actions of these compounds in the gallbladder wall.     

Immunohistochemical features of substance P-immunoreactive chromaffin cells and nerve fibers in the rat adrenal gland

The distribution of substance P (SP) immunoreactivity and the colocalization of SP with other bioactive substances in chromaffin cells and nerve fibers were investigated in the rat adrenal gland at the light microscopic level. In the capsule and cortex, SP immunoreactivity was seen in some nerve fibers around blood vessels and in thick nerve bundles passing through the cortex directly into the medulla. In the medulla, the SP immunoreactivity was observed in a small number of chromaffin cells; these SP-immunoreactive chromaffin cells were either phenylethanolamine N-methyltransferase (PNMT) immunoreactive or immunonegative, indicating that they were either adrenaline cells or noradrenaline (NA) cells. SP-immunoreactive varicose nerve fibers were also found in the medulla and were in contact with a cluster of the NA cells showing catecholamine fluorescence, which suggests that SP from medullary nerve fibers may regulate the secretory activity of the NA cells. Because no SP-immunoreactive ganglion cell was present in the rat adrenal gland, the intra-adrenal nerve fibers were considered to be extrinsic in origin. The double-immunostaining method further revealed that the SP-immunoreactive chromaffin cells also exhibit immunoreactivities for calcitonin gene-related peptide (CGRP), and neuropeptide tyrosine (NPY), suggesting that these peptides can also be released from the chromaffin cells by certain stimuli. The intra-adrenal nerve fibers in the medulla were composed of SP-single immunoreactive, and SP/CGRP-, SP/choline acetyltransferase (ChAT)-, SP/nitric oxide synthase (NOS)-, SP/pituitary adenylate cyclase activating polypeptide (PACAP)-, ChAT/NOS-, and ChAT/PACAP-immunoreactive nerve fibers, which may affect the secretory activity of the NA cells. In the adrenal capsule, the nerve fibers were present around blood vessels and showed immunoreactivities for SP/ CGRP, SP/NPY, SP/NOS, and SP/vasoactive intestinal polypeptide, suggesting that the origin of nerve fibers in the capsule may differ from those in the medulla.     

Nitric oxide synthase immunoreactivity of interstitial cells of Cajal in experimental colitis

OBJECTIVE: There is functional and morphological evidence that interstitial cells of Cajal (ICC) may play a role in nitric oxide (NO) dependent signal transduction. However, little is known about the nitric oxide synthase (NOS) containing ICC during inflammation. MATERIALS AND METHODS: Immunocytochemical methods were used for the ultrastructural localization of NOS1-containing ICC in the wall of the colon of rats in experimental colitis. RESULTS: Large numbers of NOS immunoreactive (IR) nerve terminals were found in very close vicinity to smooth muscle cells as well as to blood vessels. IR nerves were found in close relationship with the ICC. The gap between the NOS IR nerve fibers and the membrane of smooth muscle cells and of ICC was 20-250 nm. In experimental colitis the number of NOS IR nerve fibers slightly decreased, however, large numbers (24%) of the ICC became IR for NOS. In the noninflamed area and in the controls, all these cells were immunonegative for NOS. CONCLUSIONS: Our light- and ultrastructural study suggests that some of the ICC can also synthesize NO, at least during inflammation. Therefore the change in the number and structure of ICC could play an important role in the pathogenesis of a variety of motility disorders.     

Expression and distribution of GABA and GABAB‐receptor in the rat adrenal gland

The inhibitory effects of gamma‐aminobutyric acid (GABA) in the central and peripheral nervous systems and the endocrine system are mediated by two different GABA receptors: GABA_A‐receptor (GABA_A‐R) and GABA_B‐receptor (GABA_B‐R). GABA_A‐R, but not GABA_B‐R, has been observed in the rat adrenal gland, where GABA is known to be released. This study sought to determine whether both GABA and GABA_B‐R are present in the endocrine and neuronal elements of the rat adrenal gland, and to investigate whether GABA_B‐R may play a role in mediating the effects of GABA in secretory activity of these cells. GABA‐immunoreactive nerve fibers were observed in the superficial cortex. Some GABA‐immunoreactive nerve fibers were found to be associated with blood vessels. Double‐immunostaining revealed GABA‐immunoreactive nerve fibers in the cortex were choline acetyltransferase (ChAT)‐immunonegative. Some GABA‐immunoreactive nerve fibers ran through the cortex toward the medulla. In the medulla, GABA‐immunoreactivity was seen in some large ganglion cells, but not in the chromaffin cells. Double‐immunostaining also showed GABA‐immunoreactive ganglion cells were nitric oxide synthase (NOS)‐immunopositive. However, neither immunohistochemistry combined with fluorescent microscopy nor double‐immunostaining revealed GABA‐immunoreactivity in the noradrenaline cells with blue‐white fluorescence or in the adrenaline cells with phenylethanolamine N‐methyltransferase (PNMT)‐immunoreactivity. Furthermore, GABA‐immunoreactive nerve fibers were observed in close contact with ganglion cells, but not chromaffin cells. Double‐immunostaining also showed that the GABA‐immunoreactive nerve fibers were in close contact with NOS‐ or neuropeptide tyrosine (NPY)‐immunoreactive ganglion cells. A few of the GABA‐immunoreactive nerve fibers were ChAT‐immunopositive, while most of the GABA‐immunoreactive nerve fibers were ChAT‐immunonegative. Numerous ChAT‐immunoreactive nerve fibers were observed in close contact with the ganglion cells and chromaffin cells in the medulla. The GABA_B‐R‐immunoreactivity was found only in ganglion cells in the medulla and not at all in the cortex. Immunohistochemistry combined with fluorescent microscopy and double‐immunostaining showed no GABA_B‐R‐immunoreactivity in noradrenaline cells with blue‐white fluorescence or in adrenaline cells with PNMT‐immunoreactivity. These immunoreactive ganglion cells were NOS‐ or NPY‐immunopositive on double‐immunostaining. These findings suggest that GABA from the intra‐adrenal nerve fibers may have an inhibitory effect on the secretory activity of ganglion cells and cortical cells, and on the motility of blood vessels in the rat adrenal gland, mediated by GABA‐Rs.