P物质免疫反应阳性神经纤维雌性大鼠生殖器官的分布

实验采用用亲和免疫组化ＳＡＢＣ法对雌性大鼠动情间期Ｐ物质的正常进行了观察。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。     

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     

雌性大鼠生殖器官一氧化氮合酶Ⅰ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期一氧化氮合酶Ⅰ的正常分布进行了观察。一氧化氮合酶Ⅰ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,曲张体处深染不典型,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,在各生殖管道,神经纤维主要与肌束同方向走行或包绕在血管周。分析雌性大鼠生殖器官内一氧化氮合酶Ⅰ免疫反应阳性神经纤维的分布,提示一氧化氮可能与血管活性或非血管平滑肌运动调节有关。     

扬子鳄卵巢内血管活性肠肽、神经肽Y和P物质的分布

目的:观察扬子鳄卵巢内血管活性肠肽（vasoactive　intestinal　peptide,VIP）、神经肽Y（neuropeptide Y,NPY）和P物质（substance P,SP）的分布情况。方法:免疫荧光法。结果:扬子鳄卵巢内的VIP免疫反应（VIP-immunoreac-tive,VIP-IR）阳性神经纤维呈波浪状,部分包绕卵泡,大部分交织在卵泡间血管周围,并构成较密集的网络;SP和NPY免疫反应（SP-and NPY-immunoreactive,SP-和NPY-IR）阳性神经纤维均为点线状,主要沿卵泡间血管周围分布,其中,部分NPY-IR纤维也构成较密集的网络。切片中均未见到三种肽能神经元的细胞体。结论:扬子鳄卵巢内存在有VIP、NPY和SP阳性神经纤维分布,主要位于卵泡间血管周围。     

P物质和降钙素基因相关肽在大鼠垂体前叶神经纤维内的共存

为观察P物质(SP)和降钙素基因相关肽(CGRP)在大鼠垂体前叶神经纤维内的共存,用种属特异性抗体(兔抗-SP多克隆抗体和小鼠抗-CGRP单克隆抗体)进行免疫荧光双标记,激光共聚焦扫描显微镜观察进行研究。结果显示:所有的CGRP免疫阳性神经纤维都是SP免疫阳性,同时所有的SP免疫阳性神经纤维也都是CGRP免疫阳性;CGRP免疫阳性神经纤维的分布和形态与SP免疫阳性神经纤维相一致。结论:P物质和降钙素基因相关肽在大鼠垂体前叶神经纤维内完全共存。     

人皮肤经络线上P物质和VIP免疫反应神经纤维与肥大细胞关系的光镜和电镜研究

本研究在已发现神经与肥大细胞之间有突触样联接关系的的基础上,检查了与肥大细胞形成联接的神经纤维终末内是否含有P物质和血管活性肠肽(VIP)。证明:光镜下P物质免疫反应阳性神经纤维与肥大细胞直接接触;将光镜看到的目的物用原位包埋法转到电镜下,可见P物质免疫反应轴索与肥大细胞紧密并靠,两者之间距离不超过20nm。含P物质免疫反应囊泡的轴突末梢在近肥大细胞处形成膨大,有的膨大部直接与肥大细胞接触,形成突触样联接。证实了前人关于与肥大细胞形成联接的轴突末梢内含有P物质的推断。此外,在光镜下证明的P物质和VIP免疫反应神经纤维在皮肤的分布与前人的报道一致,本实验并观察了它们在电镜下的结构特点。     

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。     

神经肽Y、P物质在扬子鳄泄殖腔壁内的分布

目的 :观察含神经肽Y(NPY)、P物质 (SP)神经在扬子鳄泄殖腔壁内的分布情况。方法 :免疫组织化学ABC法及免疫荧光法。结果 :扬子鳄泄殖腔壁内SP免疫反应 (SP IR)阳性神经纤维多见于肌层 ,也见于外膜、呈细线状或点线状 ,肌层内的SP IR神经纤维与平滑肌纤维平行走行或构成网络状 ;NPY免疫反应 (NPY IR)神经纤维呈细线状 ,密度较稀 ,主要见于肌层 ,也近似与平滑肌纤维平行走行 ,免疫荧光法还证实 ,肌层内有散在分布的、多呈椭圆形的NPY IR阳性神经元胞体 ,并见有突起与周围的神经纤维形成联系。结论 :扬子鳄的泄殖腔壁也存在有SP、NPY能神经分布     

降钙素基因相关肽免疫反应神经纤维在大鼠脾的分布

用免疫组织化学ＡＢＣ法，研究了降钙素基因相关肽免疫反应神经纤维在大鼠脾内的分布，在大鼠脾内有丰富的ＣＧＲＰ免疫反应神经纤维，它们多呈串珠状，主要分布于红髓。尤其是脾血窦和一些小血管，也见于边缘区的淋巴组织中，偶见于动脉周围淋巴鞘，脾内ＣＧＲＰ免疫反应神经纤维与淋巴细胞关系密切，可能参与调节脾淋巴细胞的发育和功能。     

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。     

Met5-enkephalin- and Met5-enkephalin-Arg6-Gly7-Leu8-immunoreactive nerve fibers in the pig female reproductive system

The localization of Met5-enkephalin (ME) immunoreactivity in the female genital organs of the rat, guinea pig and pig was studied by indirect immunofluorescence method. In the rat and guinea pig, no ME immunoreactivity was observed in the uterus, fallopian tube or ovary. In the pig uterus and fallopian tube ME-immunoreactive nerve fibers were observed in muscular and submucose layers as well as around the blood vessels. In the pig ovary, ME immunoreactivity was localized in nerve fibers in medullary and cortical parts of the organ. Met5-enkephalin-Arg6-Gly7-Leu8 (MEAGL) immunoreactivity was also studied in the pig uterus, where its distribution was similar to that of ME. The present results suggest that the pig genital organs receive innervation by nerve fibers containing proenkephalin A-derived peptides, which may have a role in modulation of neurotransmission in these organs.     

Distribution of somatostatin- and neuropeptide Y-immunoreactive nerve fibers in the porcine female reproductive system.

The localization and distribution of somatostatin and neuropeptide Y were studied in the porcine female reproductive system with the indirect immunofluorescence technique. Somatostatin-immunoreactive nerve fibers were observed in different parts of the ovary and in the muscular membrane of the uterus as well as in the mesosalphinx. Somatostatin-immunoreactive neurons were detected in the inferior mesenteric ganglion. Neuropeptide Y immunoreactivity was present in a large number of nerve fibers distributed in different regions of the uterus, oviduct and ovary. The present results suggest that the porcine female genital organs receive innervation by somatostatin- and neuropeptide Y-containing nerve fibers, but their exact functional role remains to be established.     

Calcitonin gene-related peptide inhibits spontaneous contractions in human uterus and fallopian tube

The localization and effects of calcitonin gene-related peptide (CGRP) in the human uterus and fallopian tube were investigated. CGRP-immunoreactive nerve fibers were found in the muscular layers, around blood vessels and close to the epithelium. The oviduct and uterine cervix were more densely innervated than the corpus of the uterus. Substance P (SP)-like immunoreactivity was found in nerves with an overlapping distribution to that of CGRP-positive fibers. CGRP (2 X 10(-10) to 10(-7) M) dose-dependently and reversibly inhibited spontaneous contractions of uterine and oviductal strips, as well as SP-induced contractions in the oviduct. A role for CGRP-containing nerve fibers in regulation of motor activity in human female reproductive organs is suggested.     

Some nerve endings in the rat pelvic paracervical autonomic ganglia and varicosities in the uterus contain calcitonin gene-related peptide and originate from dorsal root ganglia

The pelvic paracervical autonomic ganglia of female rats were studied for a subpopulation of nerve endings that could be derived from sensory nerve fibers. Immunohistochemical staining using an antiserum against the synaptic-terminal protein synapsin I was used to identify terminal boutons, while an antiserum against the neuropeptide calcitonin gene-related peptide was used to reveal a subpopulation of sensory nerve fibers. The uterine cervix was also examined for the existence of calcitonin gene-related peptide and synapsin I immunoreactivity in nerve fiber varicosities. In addition, the location of nerve endings in the paracervical ganglion was compared to that in the superior cervical ganglion. Synapsin I immunoreactivity was present in the paracervical ganglion in abundant boutons around neuron somata and in the cervix in varicose nerve fibers of the myometrium, vasculature and epithelium. Double labeling immunocytochemistry revealed calcitonin gene-related peptide-like immunoreactivity in subpopulations of synapsin I-immunoreactive endings in ganglia and nerve varicosities in the cervix. Injection of a retrograde axonal tracer, fluorogold, into the paracervical ganglion produced labeled neurons in dorsal root ganglia and spinal cord; however, fluorogold-labeled neurons containing calcitonin gene-related peptide immunoreactivity were visualized only in dorsal root ganglia. Injections of fluorogold into the uterine cervix produced labeled neurons in the paracervical ganglion and dorsal root ganglia; however, only those in dorsal root ganglia contained immunoreactivity for calcitonin gene-related peptide. These results suggest that immunoreactivity for calcitonin gene-related peptide is present in a subpopulation of nerve endings in the paracervical ganglion and not merely in fibers of passage. The nerve endings in the ganglion and varicosities in the uterine cervix originate from sensory neurons in dorsal root ganglia. The arrangement of endings in the ganglia could play a role in sensory/autonomic interactions for modulation of visceral activity.     

Restricted innervation of uterus and placenta during pregnancy: evidence for a role of the repelling signal Semaphorin 3A.

Because data from the literature suggest a lack of innervation of the placenta, we have investigated placenta, umbilical cord, and uterus to identify the molecules that play a role in regulating innervation in these organs. Neuropilin-1 and Plexin-A1 are cell surface proteins that form a receptor complex for Semaphorin 3A (Sema 3A), a secreted molecule mediating repelling signals for axonal growth cones. We have analyzed the expression of Neuropilin-1, Plexin-A1, and Semaphorin 3A in the above-mentioned tissues on the hypothesis that these molecules could regulate innervation in these organs during gestation. We found that nervous fibers are only present in the proximal part of the umbilical cord, close to the newborn, and in nongestational uterine tissues. In contrast, nervous fibers are not present in the distal segment of the umbilical cord, in the placenta and in the uterine tissues during gestation. We also found that Sema 3A receptors, Neuropilin-1 and Plexin-A1, are expressed by the nervous fibers of the proximal part of the umbilical cord, whereas Sema 3A is secreted in the umbilical cord, in the placenta, and in gestational uterine tissues. We report that a factor secreted in the umbilical cord induces the collapse of neurite growth cones in vitro and provide evidence that this factor is Sema 3A. In summary, our results suggest that the chemorepulsive signals mediated by Sema 3A play an important role in preventing nerve fibers growth in the umbilical cord and in gestational uterine tissues. The inhibition of nerve growth into the myometrium as well as into the placenta could be considered fundamental processes to preserve the fetus from external stressful events.     

Innervation of internal female genital organs in the pig during prenatal development

This study investigated the innervation of internal genital organs in 5‐, 7‐ and 10‐week‐old female pig foetuses using single and double‐labelling immunofluorescence methods. The structure and topography of the organs was examined using a scanning electron microscope (SEM). The investigations revealed differences in the innervation between the three developmental periods. Immunostaining for protein gene product 9.5 (PGP; general neural marker) disclosed solitary nerve fibres in the external part of the gonadal ridge and just outside of the mesenchyme surrounding mesonephric ducts in 5‐week‐old foetuses. Double‐labelling immunohistochemistry revealed that nerve fibres associated with the ridge expressed dopamine β‐hydroxylase (DβH; adrenergic marker) or vesicular acetylcholine transporter (VAChT; cholinergic marker). In 7‐week‐old foetuses, the PGP‐positive nerve terminals were absent from the gonad but some of them ran outside and along, and sometimes penetrated into the mesenchyme surrounding the tubal and uterine segments of the paramesonephric ducts and uterovaginal canal. Few axons penetrated into the mesenchyme. DβH‐positive fibres were found in single nerve strands or bundles distributed at the edge of the mesenchyme. VAChT‐positive nerve terminals formed delicate bundles located at the edge of the mesenchyme, and the single nerves penetrated into the mesenchyme. DβH was also expressed by neurons which formed cell clusters comprising also DβH‐ or VAChT‐positive nerve fibres. In 10‐week‐old foetuses, PGP‐positive nerve fibres were still absent from the ovary but some were distributed in the mesenchyme associated with the uterovaginal canal and uterine and a tubal segment of the paramesonephric ducts, respectively. DβH‐ or VAChT‐positive nerve fibres were distributed at the periphery of the mesenchyme associated with the uterovaginal canal. Some DβH‐ and many VAChT‐positive nerve fibres were evenly distributed throughout the mesenchyme. The clusters of nerve cells comprised DβH‐positive perikarya and DβH‐ or VAChT‐positive nerve fibres. The investigations revealed no DβH/VAChT‐positive nerve fibres or neurons as well as no nerve structures stained for calcitonin gene‐related peptide and/or substance P (sensory markers) associated with the genital organs in the studied prenatal periods.     

Functional properties of afferent fibers supplying reproductive and other pelvic organs in pelvic nerve of female rat

1. Electrophysiological techniques were used to characterize responses of afferent fibers in pelvic nerve of adult, virgin female rats to mechanical or chemical stimulation of internal reproductive organs and to mechanical stimulation of other pelvic organs. 2. In an in vivo barbiturate-anesthetized preparation, pelvic nerve afferent fibers responded to a wide variety of mechanical stimulation applied to restricted regions of the vaginal canal, caudal uterus (body and cervix), bladder, ureter, colon, or anus. 3. Single-fiber mechanoreceptive fields were invariably confined to a single organ. Notably, responses could be evoked not only by gentle stimulation of the unit's receptive field directly on the organ itself, but also by stimulating the field indirectly with intense stimulation through the appropriate part of a contiguous organ. This innervation feature is consistent with the separability of pelvic organ functions under innocuous conditions but their confusion under noxious ones. 4. Receptive fields on the reproductive organs extended from the caudal edge of the vagina to the uterine body (including the cervix) but were most often located in the fornix (vaginocervical junction). Most units had no or low levels of spontaneous activity. Their responses to mechanical stimuli were usually slowly or moderately adapting and time-locked to the stimulus. 5. Fibers with vaginal receptive fields (including the fornix) responded best either to vaginal distension with a balloon or, more often, to a probe moving along the internal vaginal surface in a direction toward the cervix. They were observed most frequently during the proestrus stage of the rat's estrous cycle. These fibers, therefore, seem particularly suited for relaying information about stimuli that occur during mating. 6. Fibers with receptive fields on the uterine cervix and body responded best to static pressure and were observed less frequently than those with vaginal fields, regardless of estrous stage. They were, however, sensitized by hypoxia. In addition, irritation of the uterus increased the probability of observing them. These fibers, therefore, may exert their primary function during reproductive conditions different from those of virgin rats, such as parturition. 7. Response activity of most of the mechanoreceptive afferent fibers supplying reproductive organs increased as the stimulus intensity increased into the noxious range; i.e., into a range in which the stimulus momentarily produced ischemia at the stimulus site. In addition, in an in vitro preparation, pelvic nerve fibers responded in a dose-dependent manner to injections through the uterine artery of bradykinin (BRAD) as well as to other algesic chemicals, 5-hydroxytryptamine (5-HT) and KCl.(ABSTRACT TRUNCATED AT 400 WORDS)     

Effect of Reserpine on the Noradrenaline Content of the Vas Deferens and the Seminal Vesicle Compared with the Submaxillary Gland and the Heart of the Rat

A low dose (50 μg/kg) of reserpine was used in order to elucidate the possible difference in rate of noradrenaline depletion between the male genital organs, the submaxillary gland and the heart of the rat. Reserpine reduced the noradrenaline content of the male genital organs more slowly and less completely than that of the submaxillary gland and the heart. There were no differences in reserpine sensitivity between innervated and decentralized genital organs. The decentralized submaxillary gland, however, showed a tendency to less rapid initial depletion when compared with the innervated gland. It is suggested that the differences in depletion rate could be due to different properties of the short adrenergic neurons innervating the genital organs when compared with the ordinary long ones to the heart and the salivary gland. Another contributing factor may be differences in blood flow through the organs. It is further suggested that the genital organs have a lower sympathetic nerve impulse frequency in vivo.     

Topography of the pelvic autonomic nervous system and its potential impact on surgical intervention in the pelvis

Bladder, bowel, and sexual dysfunction caused by iatrogenic lesions of the inferior hypogastric plexus (IHP) are well known and commonly tolerated in pelvic surgery. Because the pelvic autonomic nerves are difficult to define and dissect in surgery, and their importance often ignored, we conducted a gross anatomic study of 90 adult and four fetal hemipelves. Using various non-surgical approaches, the anatomic relations and pathways of the IHP were dissected. The IHP extended from the sacrum to the genital organs at the level of the lower sacral vertebrae. It originated from three different sources: the hypogastric nerve, the sacral splanchnic nerves from the sacral sympathetic trunk (mostly the S2 ganglion), and the pelvic splanchnic nerves, which branched primarily from the third and fourth sacral ventral rami. These fibers converge to form a uniform nerve plate medial to the vascular layer and deep to the peritoneum. The posterior portion of the IHP supplied the rectum and the anterior portion of the urogenital organs; nerve fibers traveled directly from the IHP to the anterolateral wall of the rectum and to the inferolateral and posterolateral aspects of the urogenital organs. The autonomic supply from the IHP was supplemented by nerves accompanying the ureter and the arteries. An understanding of the location of the autonomic pelvic network, including important landmarks, should help prevent iatrogenic injury through the adoption of surgical techniques that reduce or prevent postoperative autonomic dysfunction.