Distribution of neuropeptide-like immunoreactive nerve fibers in the canine larynx.

The distribution of neuropeptide immunoreactive nerve fibers in the canine larynx was examined. In the epithelium of supra- and subglottic regions, a dense distribution of substance P (SP)- and calcitonin gene-related polypeptide (CGRP)-immunoreactive (IR) nerve fibers was observed. Some vasoactive intestinal polypeptide (VIP)-IR intraepithelial nerve fibers were also seen in the subglottic region. In the laryngeal glands, a dense distribution of VIP-IR nerve fibers with a few SP- and enkephalin (ENK)-IR nerve fibers were found around the acini. In the walls of arteries in the lamina propria, many VIP-, SP-, and CGRP-IR nerve fibers were seen, whereas neuropeptide Y-, ENK-, and VIP-IR nerve fibers were predominantly distributed around the arteries in the vocal muscle. In the free edge of the vocal cord, few immunoreactive nerve fibers were detected within the epithelium and around the arteries in the lamina propria. These results suggest that there are regional differences in the occurrence of peptides in nerve fibers innervating the epithelium and the blood vessels in the larynx and that the perception mechanism of the epithelium and the regulatory system of local blood flow are varied according to their location in the larynx.     

Substance P immunoreactive nerve fibers of the canine laryngeal mucosa

Substance P (SP) immunoreactive nerve endings in the laryngeal mucosa were studied by PAP immunohistochemistry with light and electron microscopy. SP immunoreactive sensory endings were observed in the epithelium as intra-epithelial free nerve endings and taste bud-like structures. A small number of autonomic SP immunoreactive nerve fibers were observed running parallel to arterioles which were over 30 micron in diameter and terminated in glandular cells. Contrary to findings by silver impregnation, intraepithelial free nerve endings were more frequently observed on the lower surface of the vocal cord. The taste bud-like structures were classified into two different types: simple terminations and reticular terminations, according to the mode of the SP immunoreactive nerve fiber. Immature or degenerated taste bud-like structures in the larynx were assumed to be mechanical receptors because these receptors lacked outer taste pores and taste hairs.     

慢传输型便秘大鼠结肠肌间神经丛内VIP能神经、SP能神经的免疫组化研究

目的　探讨结肠肌间神经丛血管活性肠肽 (VIP)能神经及P物质 (SP)能神经在慢传输型便秘发病中的作用。方法　建立大鼠慢传输型便秘模型 ,利用铺片技术制作结肠肌间神经丛标本 ,采用免疫组织化学法在铺片上显示便秘大鼠结肠肌间神经丛内VIP能神经及SP能神经的分布及形态学改变。结果　与正常大鼠比较 ,便秘大鼠结肠肌间神经丛VIP能神经节及节间束细小 ,节内神经元数量明显减少 ,节间束内神经纤维分布稀疏 (平均面密度值 2 3.0 8± 1.82vs 19.35± 1 13,P <0 0 1) ;便秘大鼠结肠肌间神经丛SP能神经节明显增大 ,神经束增粗 ,神经元和神经纤维明显增多 (平均面密度值 15.4 5± 1 0 5vs18.14±1 18,P <0 .0 1)。结论　慢传输型便秘大鼠结肠壁内存在明显的神经病理学改变 ,结肠传输功能障碍可能与结肠肌间神经丛VIP和SP能神经病理改变和 或功能障碍有关。     

Pharyngeal branch of the vagus nerve carries intraepithelial afferent fibers in the cat pharynx: An elucidation of the origin and central and peripheral distribution of these components

The presence of a sensory component in the pharyngeal branch of the vagus nerve (PhB), including its peripheral distribution and central projection, was studied by denervation and tracer experiments in the cat. The distribution of nerve fibers immunoreactive to protein gene product 9.5, a sensitive neuronal marker; calcitonin gene-related peptide; and substance P in the pharyngeal epithelium was analyzed in both intact animals and animals subjected to partial denervation by means of sectioning two of the three nerve trunks, the glossopharyngeal nerve, the superior laryngeal nerve, and the PhB, while leaving one intact. The results of this study show that the glossopharyngeal nerve and superior laryngeal nerve carry nerve fibers to the pharyngeal epithelium rostral and caudal to the middle level of the epiglottis, respectively, whereas the PhB carries nerve fibers to the mesopharyngeal epithelium. Tracer experiments, by applying wheat germ agglutinin-horseradish peroxidase to the PhB, demonstrated retrogradely labeled primary sensory neurons in the jugular ganglion and transganglionic labeling of terminals in the interstitial subnucleus of the nucleus of the tractus solitarius. These results indicate that the PhB contains a sensory component that originates from the jugular ganglion, innervates the mesopharyngeal epithelium, and projects to the nucleus of the tractus solitarius.     

Autonomic neuropeptides in the interface membrane of aseptic loose hip prostheses.

We analyzed the presence of autonomic nerve fibers in the interface membranes (n = 9) surrounding aseptic loosened hip prostheses by immunohistochemistry. The study focused on the autonomic messengers neuropeptide Y (NPY), tyrosine hydroxylase (TH), the rate-limiting enzyme in the synthesis of noradrenaline (NA), and vasoactive intestinal polypeptide (VIP). Protein gene product (PGP) 9.5, a general marker of peripheral nerve fibers, was also analyzed to establish the neuronal character of the immunoreactive structures. PGP 9.5-positive and NPY-positive nerve fibers were identified in all 9 samples, and VIP-immunoreactive and TH-immunoreactive fibers were found in 7. There was a difference in the distribution of nerve fibers both between and within the samples. Among the neuropeptides analyzed, NPY was most abundant. NPY-positive and TH-positive fibers were predominantly found around the blood vessel walls forming varicose nerve terminals. VIP-positive fibers were mainly observed as thin varicose nerve terminals with no relationship to blood vessels. Autonomic neuropeptides exert not only vasoactive and immunoregulatory effects, but also have been found to have direct effects on bone tissue. Moreover, the autonomic nervous system has been strongly implicated in nociception and inflammation. Neuronal NPY, TH, and VIP in the interface membrane may prove to contribute to the pathologic mechanisms leading to aseptic loosening of hip prostheses.     

Morphological basis for back pain: the demonstration of nerve fibers and neuropeptides in the lumbar facet joint capsule but not in ligamentum flavum.

The innervation of lumbar facet capsule and ligamentum flavum was investigated using antisera to a general neuronal marker protein gene product (PGP) 9.5 and to peptide markers of sensory nerves (calcitonin gene-related peptide [CGRP] and substance P) and autonomic nerves (vasoactive intestinal polypeptide [VIP] and C-flanking peptide of neuropeptide Y [CPON]). In the facet capsule (n = 14), PGP 9.5 and CGRP-immunoreactive nerves occurred in 12 and five specimens, respectively, both around blood vessels and as free fibers in the stroma. Free fibers immunoreactive for substance P or VIP were noted in three and five specimens, whereas in nine specimens there were CPON-immunoreactive nerves located perivascularly. There was no immunoreactivity in the ligamentum flavum. This study provides further evidence that the facet capsule but not the ligamentum flavum has substantial innervation by sensory and autonomic nerve fibers and has a structural basis for pain perception.     

Autonomic innervation of tendons, ligaments and joint capsules. A morphologic and quantitative study in the rat.

We analyzed the neuronal occurrence of autonomic transmitters; noradrenaline (NA), neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP), in the Achilles tendon, medial and lateral collateral ligaments and knee joint capsule in the rat--by immunohistochemistry (IHC). In addition, the tissue concentrations of the sympathetic neuropeptide, NPY, and the parasympathetic peptide, VIP, were determined by radioimmunoassay (RIA). IHC demonstrated nerve fibers containing sympathetic vasoconstrictors--NA and NPY--and the parasympathetic vasodilator, VIP, in all tissues. NPY- and NA-positive nerve fibers were predominantly observed in larger blood vessels, whereas, nerve fibers immunoreactive to VIP were found in smaller vessels. In many nerve fibers a co-localization of the transmitters was seen. RIA showed that the concentration of NPY compared to VIP was 15-times higher in ligaments and twice as high in tendons and capsules. The differences noted may reflect a difference in vulnerability to degenerative conditions. In pathological conditions, dysregulation of autonomic transmitters in hypovascularized tissues subjected to repetitive mechanical load may contribute to tissue hypoxia leading to degeneration and rupture of tendons and ligaments.     

Neuropeptide Y, calcitonin gene-related peptide, and galanin in Hirschsprung's disease: an immunocytochemical study

The aganglionic intestinal segment in Hirschsprung's disease is known to contain a reduced number of nerve fibers storing vasoactive intestinal peptide (VIP), substance P (SP), enkephalin, and gastrin-releasing peptide (GRP). In this study, nerves containing three newly described neuropeptides: neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP), and galanin were examined using immunocytochemistry. Nerve fibers displaying NPY immunoreactivity were found to be more frequent in the aganglionic than in nonafflicted ganglionic intestine. Nerve fibers storing CGRP and galanin on the other hand were roughly equally frequent but the distribution pattern differed in that the bulk of fibers in the aganglionic intestine was localized to large nerve trunks not seen in the ganglionic segment. The functional significance of these changes has yet to be defined.     

Mast cells and gut nerve development: implications for Hirschsprung's disease and intestinal neuronal dysplasia

BACKGROUND/PURPOSE: In Hirschsprung's disease (HD), the aganglionic bowel is characterized by the presence of hypertrophic nerve trunks and increased numbers of adrenergic and cholinergic nerve fibers. Intestinal neuronal dysplasia (IND), if associated with HD, occurs proximal to the aganglionic segment in HD, and is characterized by dysplasia of parasympathetic nerves, hyperganglionosis, and giant ganglia. However, the cause of such abnormalities in HD and IND is unclear. Recent reports that mast cells (MC) have been observed in direct contact with nerve fibers generally, suggest that MC are essential for nerve growth and repair. MC synthesize, store, and release nerve growth factor (NGF). NGF supports the development and functional maintenance of sympathetic and cholinergic neurons. The aim of this study was to examine the colonic distribution of MC with respect to nerves in HD and HD associated with IND. METHODS: MC and NGF were examined immunohistochemically in ganglionic, transitional, and aganglionic segments of colon from 20 patients with HD (five patients associated with IND) and 15 age-matched controls. MC were counted in each of five random fields using light microscopy (x100). RESULTS: Interestingly, aganglionic and IND segments had large numbers of MC in all layers compared with ganglionic segments in HD patients and controls (P< .0001). The number of MC in transitional segments was significantly less compared with ganglionic segments in HD patients and controls (P< .01). MC stained positively for NGF, and some were found in contact with abnormal hypertrophic nerve trunks in HD and giant ganglia in IND. CONCLUSIONS: MC may cause hypertrophic nerve trunks and giant ganglia by releasing NGF and also may be an important factor in the excessive development of cholinergic and adrenergic nerve fibers in HD and IND.     

大鼠盆神经节-膀胱神经通路的研究

目的探讨大鼠膀胱和盆神经节之间的神经通路及盆神经节内投射至膀胱的神经元的化学性质。方法正常雄性SD大鼠(250-300g)，辣根过氧化物酶(HRP)注射至膀胱，逆行神经追踪，经钨酸钠-四甲基联苯胺(TMB)-HRP法显色反应后再行免疫组织化学(胆碱乙酰转移酶ChAT、血管活性肠肽VIP、P物质SP、酪氨酸羟化酶TH)和辅酶Ⅱ(NADPH-d)酶组织化学法。结果支配膀胱的神经元多位于盆神经节主体部，大多数为ChAT(52．6％)和TH(23．3％)，少数为VIP(5．7％)，偶见一氧化氮合成酶NOS(5．6％)及SP(4．1％)。结论在盆神经节中支配膀胱的神经细胞中存在ChAT、TH、VIP、SP和一氧化氮(NO)多种神经递质。膀胱功能多因素调节，乙酰胆碱(Ach)和TH为其经典递质，神经肽(VIP，SP)和NO在其中起重要作用，盆神经节是排尿反射的各级中枢的最后通路。     

Origin of the adrenergic nerve fibers in the subdiaphragmatic vagus in the dog

The vagi at the subdiaphragmatic level were studied by the Hillarp-Falck technic in combination with a nerve crush procedure in three normal dogs and in eight dogs that had undergone previous surgical excision of the superior cervical ganglion and/or the stellate ganglia. Unilateral ganglionectomies were performed so that the contralateral vagus served as a control. Based on these results, it is concluded that: (1) the subdiaphragmatic canine vagus contains numerous adrenergic nerve fibers; (2) the main portion of these vagal adrenergic fibers arises from the stellate ganglia; and (3) removal of both the stellate and the superior cervical ganglia results in nearly complete adrenergic denervation of the abdominal vagus.     

背根神经节内P物质、降钙素基因相关肽免疫阳性神经元与阴茎包皮系带感觉信息的传递

目的:探讨背根神经节(DRG)内P物质(SP)、降钙素基因相关肽(CGRP)免疫阳性神经元与阴茎包皮系带感觉信息传递之间的关系。方法:通过荧光金(FG)逆行标记对大鼠阴茎包皮系带内神经末梢的来源作追踪定位,并结合SP、CGRP免疫荧光标记法,研究大鼠DRG内FG标记阳性神经元中SP、CGRP免疫阳性神经元的形态和分布。结果:FG逆行标记结果发现,大鼠阴茎包皮系带内的神经末梢起源于第6腰髓对应的背根神经节(L6-DRG)和第1骶髓对应的背根神经节(S1-DRG)的神经元。对这些神经元分别作SP、CGRP免疫荧光标记后发现,标记细胞大小不等,分别呈深红色和深绿色,沿神经束成行排列或散在分布。FG/SP、FG/CGRP双标记阳性细胞均为中小型,其数量分别占FG逆行标记阳性细胞总数的1/3和1/2,FG/SP/CGRP三标记阳性细胞占FG逆行标记阳性细胞总数的1/5。结论:大鼠L6-DRG和S1-DRG内的SP、CGRP免疫阳性神经元可能参与阴茎包皮系带感觉信息的传递。     

Peptidergic innervation in infantile hypertrophic pyloric stenosis

The gastrointestinal tract harbors several populations of peptide containing nerve fibers. Among the gut neuropeptides are vasoactive intestinal peptide (VIP), substance P, enkephalin, and gastrin releasing peptide (GRP). We have examined specimens from five patients with pyloric stenosis and from five controls immunocytochemically with respect to the density of nerve fibers containing VIP, substance P, enkephalin, or GRP. In the control specimens VIP and enkephalin fibers were fairly numerous, whereas substance P and GRP fibers were few. In the pyloric stenosis patients the density of VIP fibers and enkephalin fibers was reduced in the smooth muscle. In the myenteric ganglia there was no such reduction. Substance P and GRP fibers were rare as in controls. The results indicate a reduction of VIP and enkephalin fibers in smooth muscle in pyloric stenosis patients and may be interpreted to support the view that an impaired neuronal function is involved in the pathophysiology of pyloric stenosis.     

Reinnervation of syngeneic pancreatico-duodenal grafts in rats

BACKGROUND: Knowledge on the reinnervation of transplanted organs is scarce, and the aim of the study was therefore to evaluate to what degree syngeneic pancreas grafts were reinnervated in rats. METHODS: Syngeneic pancreatico-duodenal transplantations were performed in normoglycemic Wistar-Furth rats. Native and transplanted pancreas and duodenum were removed 4 or 40 weeks after implantation, and processed for indirect immunofluorescence using antibodies directed against vasoactive intestinal peptide, substance P (SP), calcitonin gene-related peptide (CGRP), neuropeptide Y (NPY), tyrosine hydroxylase (TH), or the general neuronal marker protein gene product 9.5. RESULTS: Four weeks after transplantation a moderate to rich number of protein gene product 9.5-positive nerve fibers were found homogeneously distributed through the pancreas, probably representing the intrapancreatic nervous system, because the grafted pancreas lacked both a sympathetic (TH/NPY) and sensory (SP/CGRP) innervation 4 weeks after implantation. In a few of the animals there was a marked increase in SP-immunoreactive nerves (lacking CGRP), most conspicuous in the duodenal portion, both 4 and 40 weeks after transplantation probably secondary to a chronic pancreatitis. The fibers seemed to emanate from intrapancreatic ganglia and possibly also from enteric neurons in adjacent parts of the duodenum. A few scattered vasoactive intestinal peptide-containing nerve fibers probably also emanating from local ganglia could be seen throughout the grafted pancreas both 4 and 40 weeks after transplantation. At 40 weeks after transplantation sympathetic (TH- and NPY-positive) nerve fibers were regularly seen, whereas CGRP-positive nerve fibers were still virtually lacking in the pancreas. To trace the origin of the ingrowing nerve fibers, the tracer True Blue was injected into the grafted pancreas of some rats 38 weeks after transplantation, i.e., 2 weeks before killing. True Blue-labeled nerve cell bodies were numerous in the celiac ganglion (presumably sympathetic nerves) and few in dorsal root ganglia (sensory nerves). CONCLUSIONS: The data suggest that the transplanted rat pancreas becomes reinnervated by mainly sympathetic nerve fibers.     

Immunohistochemical studies on the distribution of nerve fibers in the human prostate with special reference to the anterior fibromuscular stroma

BACKGROUND: Although the anterior fibromuscular stroma (AFMS) comprises up to one third of the total bulk of the prostate, its physiological function remains unknown. We recently reported the possible contribution of the AFMS to micturition. The aim of this study is to reveal the differences in the distribution of innervation between the AFMS and the other regions of the prostate. METHODS: We performed immunohistochemical stainings using antibodies such as protein gene product (PGP) 9.5, tyrosine hydroxylase(TH), neuropeptide Y (NPY), and vasoactive intestinal peptide (VIP). Morphometric analysis was made to examine the density of peripheral nerve fibers containing PGP 9.5, TH, NPY, and VIP by using a computer-assisted imaging system. RESULTS: The number of PGP 9.5-immunoreactive (IR) nerve fibers and the smooth muscle in the AFMS decreased from the base to the apex of the prostate. TH-IR nerve fibers were more abundant in the AFMS than in the transition zone. NPY- and VIP-IR nerve fibers were less numerous in the AFMS than in the peripheral zone. CONCLUSIONS: This study is the first to demonstrate that the AFMS has peculiar neuronal innervation. We observed significantly different innervation in the AFMS compared with the other regions of the prostate.     

Release of vasoactive intestinal peptide in the central nervous system in man.

Recent work has shown that vasoactive intestinal peptide (VIP), one of the many candidate hormones of the gut, also occurs widely in neurones. To determine whether the neuronal peptide may have a neurotransmitter function, we studied changes in immunoreactive VIP in dog plasma and human cerebrospinal fluid after the infusion of choline esterase inhibitors (neostigmine and physostigmine, respectively). Immunoreactive VIP was released in both situations. The systemic changes (in VIP levels) were enhanced five weeks after portacaval shunting in dogs. Our results demonstrate that the immunoreactive VIP level increases as a result of choline esterase inhibitors. The plasma "release" may originate either from peripheral peptidinergic nerve terminals or from APUD cells of the gastroenteropancreatic system. The increase in immunoreactive cerebrospinal fluid VIP may very well originate from central neurons, since the peptide does not apparently cross the blood-brain barrier.     

感觉神经(元)对失神经骨骼肌超微结构保护作用的实验研究

目的观察感觉神经（元）对失神经骨骼肌超微结构的保护作用。方法60只SD大鼠,按手术先后顺序随机分成10组,每组6只。A组(对照组)肱二头肌完全失神经支配。B～E组(实验组)B组为失神经支配加感觉神经种植组,C组为失神经支配加感觉神经寄养组,D组为失神经支配加背根神经节种植组,E组为节前撕脱的感觉神经种植组。术后1、3个月取材,各组又分为二个时间组。用透射电镜观察肌肉超微结构的变化。结果与对照组相比,各实验组肌肉退变核数少、线粒体肿胀及肌质网扩张程度轻、毛细血管/肌纤维数比值大、间质胶原纤维和成纤维细胞少、肌丝肌节排列整齐。结论感觉神经（元）对失神经骨骼肌超微结构有保护作用。     

Immunohistochemical studies of putative neurotransmitters in the lower urinary tract

Putative neurotransmitters of the lower urinary tract were investigated in apes, rabbits and cats using immunohistochemical techniques of PAP (Peroxidase antiperoxidase) staining and IGSS (Immunogold silver staining) methods for Neuron specific enolase (NSE), Acetylcholine (Ach), Noradrenaline (NA), Vasoactive intestinal polypeptide (VIP), Substance P (SP) and Calcitonin gene related peptide (CGRP). 1) The localization of pelvic ganglions exhibited more striking evidence of species difference. Huge pelvic ganglions were found particularly in the dorsolateral area of the prostate in apes. On the other hand, in cats and rabbits, many ganglion cells were found around the uretero-vesical junctions. 2) In the pelvic ganglions of the apes, Ach immunoreactives were detected in nearly 70 percent of the cell bodies. 10-15 percent were NA immunoreactive cells. In addition, 15-20 percent VIP and a smaller percentage of SP immunoreactive cells were detected in the same ganglions. Axons extending from the ganglion cells showed the intense neurotransmitters immunoreactivity. 3) In the apes, varicose fibers containing SP were widely distributed in the epithelium, submucosa, muscle layer, and around the vessels of the bladder. SP immunoreactive cell bodies were found in the dorsal root ganglion at levels of L7, S1 and at the same levels in the posterior horn. On the other hand, the bulbourethral gland and the seminal vesicle contained SP immunoreactive cell bodies. 4) CGRP containing fibers were distributed in similar locations as SP containing fibers in the bladder. 5) VIP immunoreactive fibers were also widely distributed, being most dense at the base of the bladder.(ABSTRACT TRUNCATED AT 250 WORDS)     

NPY hyperinnervation in Hirschsprung's disease: both adrenergic and nonadrenergic fibers contribute.

In Hirschsprung's disease, the aganglionic bowel is characterized by an absence of ganglion cells and an increased number of adrenergic and presumed cholinergic nerve fibers. In addition, a severe derangement of peptide-containing nerve fibers is encountered including a hyperinnervation of neuropeptide Y (NPY)-containing fibers. Using immunochemical and immunocytochemical methods, we examined the nature of the NPY-containing nerve fibers contributing to the hyperinnervation. The concentration of NPY was markedly increased in the aganglionic segment. Coexistence of NPY, vasoactive intestinal peptide (VIP), and the adrenergic enzyme tyrosine hydroxylase (TH) showed small populations of nerve fibers containing NPY/TH, NPY/VIP, or TH alone in ganglionic intestine. Numerous nerve fibers stored VIP but lacked NPY. These fibers did not contain TH, indicating that all VIP-containing fibers are nonadrenergic. In the aganglionic intestine there was a marked increase in the number of nerve fibers storing NPY/TH and NPY/VIP, whereas the fibers storing VIP alone were reduced in number. A small number of nerve fibers storing NPY alone occurred in the hypertrophic nerve bundles. NPY/VIP-containing nerve fibers were particularly numerous in the mucosa in aganglionic intestine, which may be of interest in the diagnosis of Hirschsprung's disease allowing the use of mucosal biopsy specimens. Thus, the proliferating NPY-containing nerve fibers in the aganglionic intestine seem to comprise three different populations, one adrenergic and two nonadrenergic, one of which contains in addition VIP.