The distribution of acetylcholinesterase(AChE)-positive nerves in chicken pancreas demonstrated by light and electron microscopy

The distribution of acetylcholinesterase (AChE)-positive nerve fibers in the chicken pancreas was investigated with histochemical methods at the light and electron microscopic level. AChE-positive nerve bundles were found to run along the pancreaticoduodenal artery and their branches proceed into interlobular connective tissue, form a plexus around the interlobular secretory ducts and small arteries, and penetrate the exocrine parenchyma. Intrapancreatic ganglia showing a strong AChE activity were detected within the interlobular connective tissue or between acini. The exocrine pancreas was richly innervated with AChE-positive terminals which contained a few large dense-cored vesicles (about 100 nm in diameter) and many small clear vesicles (about 50 nm in diameter). Such terminals made contact with intercalated ductular cells and the smooth muscles of larger blood vessels. The endocrine pancreas was supplied with fewer nerves than the exocrine pancreas. A different distribution of AChE-positive fibers was noticed between A- and B-islets which were distinguished immunohistochemically. B- and D-cells were richly innervated by AChE-positive nerves, whereas A-cells, only poorly. These observations make clear that the cholinergic system relates to the regulation of both exocrine and endocrine tissues, except A-cells, in the chicken pancreas.     

Neurochemical properties of the middle cervical ganglion in the sheep

The neurochemical properties of the ovine middle cervical ganglion (MCG) were studied using antibodies raised against tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DbetaH), neuropeptide Y (NPY), substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and galanin (GAL). Double-labelling immunocytochemistry revealed that the vast majority (95.5 +/- 0.8%) of postganglionic sympathetic MCG neurons expressed simultaneously both catecholamine-synthesizing enzymes (neurons were TH/DbetaH-positive). A large population of noradrenergic neurons exhibited immunoreactivity (IR) to NPY (62.2 +/- 2.2%), but single NPY-positive perikarya-lacking noradrenergic markers were also observed (2.0 +/- 0.3%). None of the examined MCG neuronal somata contained SP, CGRP, GAL or VIP. A moderate number of noradrenergic nerve fibres located amongst neuronal cell bodies was also found. In small number of these terminals the presence of NPYor GAL (but not CGRP or VIP) was detected. The ovine MCG was numerously innervated with SP-immunoreactive nerve fibres which sometimes formed basket-like formations around postganglionic neurons. The MCG exhibited a sparse CGRP-immunoreactive innervation and lacked VIP-positive nerve terminals. In many aspects the chemical coding of MCG postganglionic neurons and nerve terminals resembles that found in other mammalian cervico-thoracic paravertebral ganglia, but some important species-dependent differences exist. The functional implications of these differences remain to be elucidated.     

Distribution of CGRP, substance P, VIP and somatostatin immunoreactive nerve fibers in the internal gills of larvae of the bullfrog, Rana catesbeiana.

CGRP, substance P (SP), VIP and somatostatin were demonstrated in the internal gills of larval bullfrogs using the immunohistochemical peroxidase-antiperoxidase method. Three groups of immunoreactive fibers were distinguished by their distribution area. The first group includes CGRP, SP and VIP immunoreactive fibers running in the connective tissue of the gill tufts. These fibers were associated with the capillaries. Some CGRP fibers were distributed similarly to SP fibers, and the density of VIP immunoreactive fibers was low. The second group includes CGRP, SP, VIP and somatostatin fibers distributed in the connective tissue surrounding the ceratobranchialia. The third group includes CGRP, SP and somatostatin fibers located in the branchial muscle. Their terminals appeared to be neuromuscular junctions. No immunoreactivity of leu- or met-enkephalins was found in the internal gills. From these findings, the first group of fibers is presumed to be involved in modulating ion transport of the internal gills. The second group of fibers except for the somatostatin fibers is thought to be continuations of the first group of fibers. The third group of fibers may possibly be involved in the modulation of transmissions at the neuromuscular junction. It is unclear whether somatostatin terminals are also involved in this.     

Target-specific projections of intrinsic ganglionic neurons with different chemical codes in the canine larynx.

The distribution and origin of peptide-containing intrinsic nerve fibers within the larynx were examined by immunohistochemistry and denervation experiments in the dog. In the normal larynx, a dense network of vasoactive intestinal polypeptide (VIP)-immunoreactive (IR) fibers was seen around the acini of submucosal glands. VIP-, substance P (SP)-, or calcitonin gene-related peptide (CGRP)-IR fibers were seen in the walls of submucosal arteries, and VIP-, neuropeptide Y (NPY)-, or enkephalin (ENK)-IR fibers were seen around the arteries in the muscle tissue. Most of these peptide-IR fibers remained after bilateral denervation of the superior and inferior laryngeal nerves. Several small intrinsic ganglia were found along the peripheral branches of the laryngeal nerves. About 97% of the ganglionic neurons were VIP-IR; of these, 44% were immunoreactive to VIP alone, 22% to VIP and NPY, 13% to VIP and SP, 7% to VIP and ENK, and 14% to VIP, NPY and SP. These results reveal that the exocrine glands and blood vessels are innervated by the intrinsic ganglionic neurons and that subpopulations of ganglionic neurons with different chemical codes innervate specific target organs in the canine larynx.     

Co-localization of vasoactive intestinal polypeptide and neuropeptide Y immunoreactivities in the nerve fibers of the rat adrenal gland

The co-localization of Vasoactive Intestinal Polypeptide (VIP) with Neuropeptide Y (NPY) or its C-flanking peptide (C-PON) was investigated with immunocytochemistry methods in the adrenal gland of the rat. Most of the VIP immunoreactive (+) nerve fibers found in the capsule/glomerular zone also exhibited NPY or C-PON immunoreactivity (IR). We found that at least two populations of VIP varicose nerve fibers can be observed, the most prevalent exhibited both VIP/NPY or VIP/C-PON IR and the other which was rather scarce lacked NPY or C-PON IR. In the superficial cortex VIP/NPY or VIP/C-PON IR nerve fibers were often associated with capsular or subcapsular vascularization and extended into the zona glomerulosa. In the deeper layers of the adrenal cortex radial fibers were closely associated with the inner vascularization of the zona fasciculata and reticularis. In the adrenal medulla NPY or C-PON immunoreactivity was associated with ganglion neurons as well as chromaffin cells; these last cells were always VIP (-). VIP and NPY/C-PON IR could be co-localized in catecholaminergic nerve terminals of the adrenal cortex but not in the adrenal medulla.     

Origin and peptide content of nerve fibers in the nasal mucosa of rats

Injection of the retrograde neuronal tracer True blue into the anterior-lateral part of the nasal mucosa of rats labeled nerve cell bodies in the superior cervical ganglion, the sphenopalatine ganglion, the otic ganglion and the trigeminal ganglion on the ipsilateral side. In the superior cervical ganglion, the sphenopalatine ganglion and the trigeminal ganglion on the contralateral side, very few nerve cell bodies were labeled, indicating that these ganglia provide minor contributions only. The number of labeled cell bodies indicates that the superior cervical ganglion, the sphenopalatine ganglion and the trigeminal ganglion contribute most to the innervation of the nose, while the contribution from the otic ganglion is minor. Cell bodies in the superior cervical ganglion harbored noradrenaline (NA) or NA/neuropeptide Y (NPY); in the sphenopalatine ganglion vasoactive intestinal peptide (VIP) or VIP/NPY; in the otic ganglion VIP, VIP/NPY or VIP/substance P (SP) and in the trigeminal ganglion calcitonin gene-related peptide (CGRP) or CGRP/SP. The results from denervations and tracer experiments suggest that all NA-containing and the majority of NPY-containing fibers in the nasal mucosa are derived from the superior cervical ganglion (sympathetic nerve supply). VIP- and VIP/NPY-containing fibers originate from the sphenopalatine and otic ganglia (parasympathetic nerve supply). Nerve fibers containing CGRP and CGRP/SP emanate from the trigeminal ganglion (sensory nerve supply).     

Co-existence of neuropeptides and differential inhibition of vasodilator responses by neuropeptide Y in guinea pig uterine arteries

The co-localization of substance P (SP) with calcitonin gene-related peptide (CGRP), and vasoactive intestinal peptide (VIP) with neuropeptide Y (NPY) of the guinea pig uterine artery were investigated with immunocytochemistry. The SP/CGRP fibre population was distinct from the VIP/NPY fibre population. Both types of fibres ran in the medial-adventitial border, and appeared as coarsed and fine varicosed. Uterine arterial dilatation was evoked by acetylcholine (ACh), SP, CGRP, and VIP in precontracted arteries as examined by a sensitive in vitro method. Strong relaxations were seen by ACh, CGRP and VIP. NPY had no relaxant effect per se but was found to be a potent inhibitor of vasodilation induced by ACh and SP, while relaxations induced by VIP and CGRP were unaffected. The functional significance of NPY in the uterine artery may to a large extent be to increase tension not only by potentiation of contraction but additionally by inhibiting vasodilator responses.     

Characterization of Meibomian gland innervation in the cynomolgus monkey (Macaca fascicularis)

To characterize the innervation of the cynomolgus monkey (Macaca fascicularis) Meibomian (tarsal) glands, upper lids of six cynomolgus monkeys were investigated with electronmicroscopical and double-labeling immunocytochemical methods. Antibodies against calcitonin gene-related peptide (CGRP), dopamine-β-hydroxylase (DBH), neuropeptide Y (NPY), nitric oxide synthase (NOS), protein gene product 9.5 (PGP 9.5), substance P (SP), tyrosine hydroxylase (TH), and vasoactive intestinal peptide (VIP) were used. In addition, sections were processed for NADPH-diaphorase (NADPH-d) histochemistry. Staining for PGP 9.5 and electron microscopy showed that Meibomian gland acini were surrounded by a network of unmyelinated nerves and terminal varicose axons. The terminals contained small agranular (30–60 nm) and large granular vesicles (65–110 nm), and were observed in close contact with the basal lamina of the acini, but never internally to the basal lamina. Meibomian axons showed like-immunore-activity (LI) for the neuropeptides SP, CGRP, NPY, and VIP. In addition, the axons stained for TH, DBH, NOS, and NADPH-d. VIP-LI, NOS- and NADPH-d-positive axons appeared to be more numerous, TH- and DBH-positive axons more rare than others. Most SP-LI axons were double-labelled for CGRP-LI, some for VIP-LI or NPY-LI. In addition, some VIP-LI axons were double-labeled for NPY-LI. NPY/VIP-LI and NPY/SP-LI axons were only observed close to the Meibomian acini. Conversely, NPY-LI colocalized with TH-IR or DBH-IR predominated in perivascular nerves of Meibomian gland vasculature. The close association of varicose axons with the acini of Meibomian glands indicates that nervous signals modulate meibomian secretion. Meibomian gland nerve fibers in the cynomolgus monkey appear to utilize various neuropeptides, catecholamines and nitric oxide as transmitter substances, and seem to derive from the pterygopalatine, superior cervical and trigeminal ganglion respectively.     

Characterization of Meibomian gland innervation in the cynomolgus monkey (Macaca fascicularis)

To characterize the innervation of the cynomolgus monkey (Macaca fascicularis) Meibomian (tarsal) glands, upper lids of six cynomolgus monkeys were investigated with electronmicroscopical and double-labeling immunocytochemical methods. Antibodies against calcitonin gene-related peptide (CGRP), dopamine--hydroxylase (DBH), neuropeptide Y (NPY), nitric oxide synthase (NOS), protein gene product 9.5 (PGP 9.5), substance P (SP), tyrosine hydroxylase (TH), and vasoactive intestinal peptide (VIP) were used. In addition, sections were processed for NADPH-diaphorase (NADPH-d) histochemistry. Staining for PGP 9.5 and electron microscopy showed that Meibomian gland acini were surrounded by a network of unmyelinated nerves and terminal varicose axons. The terminals contained small agranular (30–60 nm) and large granular vesicles (65–110 nm), and were observed in close contact with the basal lamina of the acini, but never internally to the basal lamina. Meibomian axons showed like-immunore-activity (LI) for the neuropeptides SP, CGRP, NPY, and VIP. In addition, the axons stained for TH, DBH, NOS, and NADPH-d. VIP-LI, NOS- and NADPH-d-positive axons appeared to be more numerous, TH- and DBH-positive axons more rare than others. Most SP-LI axons were double-labelled for CGRP-LI, some for VIP-LI or NPY-LI. In addition, some VIP-LI axons were double-labeled for NPY-LI. NPY/VIP-LI and NPY/SP-LI axons were only observed close to the Meibomian acini. Conversely, NPY-LI colocalized with TH-IR or DBH-IR predominated in perivascular nerves of Meibomian gland vasculature. The close association of varicose axons with the acini of Meibomian glands indicates that nervous signals modulate meibomian secretion. Meibomian gland nerve fibers in the cynomolgus monkey appear to utilize various neuropeptides, catecholamines and nitric oxide as transmitter substances, and seem to derive from the pterygopalatine, superior cervical and trigeminal ganglion respectively.     

The innervation of the synovium of the knee joint in the guinea pig: an immunohistochemical and ultrastructural study

 The innervation of the knee joint synovial membrane of the guinea pig, i.e., the synoviocyte layer, the subjacent connective tissue and the connective tissue region beneath, was analyzed with immunohistofluorescence and electron microscopy. A screening of the innervation with antibodies against the general axon marker – protein gene product (PGP) 9,5 – revealed the presence of nerve fibers distributed in various regions of the knee joint synovial membrane. Confirmating previous studies, some of these nerve fibers stained with antibodies to tyrosine hydroxylase (TH), neuropeptide Y (NPY), substance P (SP), calcitonin gene-related peptide (CGRP), and vasoactive intestinal polypeptide (VIP). In addition, dynorphin (DYN)-containing fibers were detected, which have not been reported previously in normal joints. In general, the immunoreactive fibers were observed close to the synoviocytes and at blood vessels. Fibers with colocalization of NPY- and TH-like immunoreactivities (LIs), as well as of DYN- and TH-LIs were demonstrated. In the electron microscope, bundles of unmyelinated fibers as well as single fibers were found in the connective tissue region below the synoviocytes. Varicose parts of the nerve fibers contained mainly small, clear vesicles. Small and large dense-cored vesicles were also seen, but less frequently. Denser portions of the plasma membranes of some axons were observed in these regions, facing the extracellular space. Myelinated fibers were also observed in some nerve bundles. These findings emphasize the complex innervation of the synovial membrane, with nerve fibers containing a host of neuroactive substances. Altogether, these fibers are probably involved in many functions such as vasoregulation and control of synovial secretion in addition to being a source of mediators in joint inflammation. Accepted: 22 November 1997     

Morphological evidence of sensory neurons in the root of the rat tongue

In our previous studies, a large number of substance P (SP)-immunoreactive (IR) nerve fibers were detected in the rat tongue and their number increased after inflammation, suggesting that these fibers might be involved in the axon reflex. Therefore, in this study, we have examined the different neuropeptide-containing nerve elements by light, electron, and confocal laser microscopy. SP, vasoactive intestinal polypeptide (VIP), and neuropeptide Y (NPY) IR varicose fibers were numerous compared with other ones. Small groups of ganglia with perikarya IR for SP, VIP, NPY, galanin, and somatostatin were observed. The SP-IR nerve cell bodies were mainly located in the tunica propria just below the epithelial lining. Double-labeling immunohistochemistry showed that the intrinsic SP-IR neurons did not colocalize VIP. The SP containing nerve terminals were observed in and below the epithelium as well as in very close contact to or making real synapses with other neurons in the intralingual ganglion. Our data confirmed the possibility of intrinsic sensory neurons, which might be the afferent branch of the intralingual reflex arch, while the VIP- and NPY-IR neurons located in the salivary glands, around the blood vessels, and in the muscle layer might constitute the efferent site of this reflex.     

Peptidergic innervation in the sinus hair follicles of several mammalian species]

In this study, the detailed distribution of peptidergic nerve fibers in the sinus hair follicle was immunohistochemically investigated by the avidin biotin-complex method using antibodies against calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal polypeptide (VIP) and neuropeptide Y (NPY) in the cat, dog, hamster, rat, mouse and guinea pig. The peptide-immunoreactive (IR) nerve fibers presented a fine varicose appearance. They entered the sinus hair follicle after penetrating the capsule (Cap) at various levels. Peptide-IR fibers in the sinus hair were classified into the 3 types described below based on their course of entry and terminal distribution. Type A: these fibers arrive at the orifice of the sinus hair follicle from superficial dermis and innervate the upper portion of the follicle. Most of them form a network around the vibrissal shaft (VS) in the outer and inner conical body (OCB and ICB, respectively), and some are distributed around the rete ridge collar (RRC) and the sebaceous gland (SG). Type B: these fibers enter the lower third of the sinus hair follicle after forming a nerve bundle together with myelinated fibers, or accompany an artery. After distribution in the trabeculae (Trab) of the cavernous sinus (CS), they form a dense plexus in the connective tissue follicle (CTF) at the level of the CS. Some ascend through the CTF and terminate at the level of the ring sinus (RS). Type C: these fibers enter the sinus hair follicle at its base. They innervate the hair papilla (HP) and the CTF of the hair bulb. CGRP and SP were detected in all types of nerve fibers in all species investigated. These peptidergic nerve fibers showed the same distribution pattern, but CGRP fibers were more numerous than SP fibers. They were distributed at high density in the OCB, ICB, CTF, ringwulst (Rw), the Trab of the CS and the HP. A moderate number of VIP- and NPY-IR fibers, mainly types B and C, were detected in these portions in the cat, dog and hamster, but few fibers were observed in other portions or in other species. Although the basic structures of the sinus hair follicle presented almost the same features as in the mammalian species, the connective tissue of the HP extended to the level of the RS in the cat, rat, hamster, and guinea pig. In these species, CGRP-, SP- and/or VIP-IR fibers extended to the top of the papillary connective tissue, and in the cat, were especially well developed.(ABSTRACT TRUNCATED AT 400 WORDS)     

人胰结缔组织纤维的细微和超微构筑及其意义--NaOH消蚀法SEM观察

目的探讨胰结缔组织纤维的细微和超微构筑及其意义.方法采用NaOH消蚀和超声波(USW)清洗技术,制备只保留结缔组织纤维成分的人胰组织样品,进行扫描电镜(SEM)观察和分析.结果经NaOH消蚀和USW清洗处理的胰组织,其细胞和基质成分全被消蚀掉,只留有结缔组织纤维,仍保持原有构筑特征.胰腺小叶、小叶间结缔组织和胰岛的轮廓清晰可见.穿行于小叶间结缔组织内的胰腺导管、动脉和静脉血管、淋巴管以及神经纤维束,易被辨认和区别.结缔组织纤维织成的束或网,均由胶原蛋白聚合而成的纳米(10-100mm)纤维,或称胶原细纤维所组成.结论活体上充满组织液的胰结缔组织纤维网,构成了该器官的组织支架和通道,为胰内、外分泌细胞的支持与保护、营养与代谢、分泌与通讯、再生与修复等,提供了适宜的微环境.     

Markers for vitiligo related neuropeptides in human skin nerve fibers

Skin distribution of substance P (SP)-, somatostatin (SOM)-, calcitonin-gene-related peptide (CGRP)- and neuropeptide-Y (NPY)-like immunoreactivity in vitiligo patients was studied by an indirect immunofluorescence technique. Immunocytochemical characteristics of the epidermis, dermoepidermal junction, papillary and reticular dermis, and skin appendages were analyzed in lesional and marginal vitiligo areas as well as in healthy skin. SP-, SOM-, CGRP-, and NPY-immunoreactive nerve fibers were observed in healthy pigmented skin, with patterns specific for immunoreactive distribution. Thin SP-containing fibers were observed in dermal papillae, extending into the epidermis, and SP-immunoreactive nerve fibers were seen around blood vessels and sweat glands. SOM-immunoreactive varicose nerve fibers were associated with Meissner's corpuscles in dermal papillae, while CGRP-like immunoreactivity was demonstrated in free subepidermal nerve terminals and sensory nerve fibers around blood vessels, hair follicles and sweat glands. Autonomic NPY-containing nerve fibers innervated eccrine sweat glands and blood vessels. The distribution of these neuropeptides was the same in healthy controls, except for an increased immunoreactivity to NPY and to a lesser extent to CGRP. These results suggest that NPY may serve as a neurochemical marker in the pathogenesis of the disease, thus supporting the neuronal theory of vitiligo.     

Immunohistochemical localisation of regulatory neuropeptides in human circumvallate papillae

The occurrence and distribution of neuropeptide-containing nerve fibres in the human circumvallate papillae were examined by the peroxidase–antiperoxidase immunolocalisation method using surgical specimens that had not been subjected to radiotherapy, and the abundance of neuropeptide-containing fibres was expressed as the percentage of total nerve fibres demonstrated by protein gene product (PGP) 9.5 immunoreactivity for a quantitative representation of these peptidergic fibres. Substance P (SP) and calcitonin gene-related peptide (CGRP) immunoreactive (IR) nerve fibres were densely distributed in the connective tissue core of the circumvallate papillae, and some SP and CGRP-IR fibres were associated with the taste buds. A moderate number of vasoactive intestinal polypeptide (VIP)-IR fibres and a few galanin (GAL)-IR fibres were also seen in the connective tissue core and subepithelial layer. There were, however, no VIP-IR or GAL-IR fibres associated with the taste buds. Neuropeptide Y (NPY)-IR fibres were few and were associated with the blood vessels. Within the epithelium of the circumvallate papillae, no peptidergic fibres were found, although a number of PGP 9.5-IR fibres were detected. The abundance of SP, CGRP, VIP, and GAL-IR fibres expressed as the percentage of total PGP 9.5 IR fibres was 25.35±3.45%, 22.18±3.26%, 10.23±1.18%, and 4.12±1.05%, respectively. The percentage of NPY-IR fibres was below 3%. In a deeper layer of the papillae, a few VIP, GAL, and NPY-IR ganglion cells were found, and VIP immunoreactivity was detected in a few cells of the taste buds. There was no somatostatin, leucine enkephalin, or methionine enkephalin immunoreactivity in the circumvallate papillae. These results suggest that the dense SP and CGRP-IR fibres within the connective tissue core of the human circumvallate papillae may be involved in the deep sensation of the tongue.     

Immunohistochemical characteristics of human paraganglion cells and sensory corpuscles associated with the urinary bladder. A developmental study in the male fetus, neonate and infant

Triple label immunohistochemistry was used to study the coexistence of the catecholamine-synthesising enzymes dopamine beta-hydroxylase (DBH) and tyrosine hydroxylase (TH) and several neuropeptides including neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), substance P (SP), calcitonin gene-related peptide (CGRP), somatostatin (SOM) and galanin (GAL) as well as nitric oxide synthase (NOS) in developing pelvic paraganglion cells in a series of human male fetal, neonatal and infant specimens ranging in age from 13 wk of gestation to 3 y postnatal. 13–20 wk old fetal specimens possessed large clusters of paraganglion cells lying lateral to the urinary bladder and prostate gland which were intensely DBH-immunoreactive (-IR) but lacked TH, NOS and the neuropeptides investigated. With increasing fetal age small clusters of paraganglion cells were observed in the muscle coat of the urinary bladder. At 23 wk of gestation occasional paraganglion cells were NOS or NPY-IR while at 26 wk of gestation the majority of paraganglion cells were TH-IR and a few were SOM or GAL-IR. Some postnatal paraganglia within the bladder musculature contained cells which were all VIP, SP or CGRP-IR while others displayed coexistence of NOS and NPY, SP and CGRP, or NPY and VIP. The presence of NOS in certain paraganglion cells indicates their capacity to generate nitric oxide (NO). These results show that human paraganglion cells develop different phenotypes possibly dependent upon their location within the bladder wall. A delicate plexus of branching varicose nerves was observed in the fetal paraganglia which increased in density with increasing gestational age. The majority of these nerves were VIP-IR while others were CGRP, SP, NPY, NOS or GAL-IR. The presence of nerve terminals adjacent to the paraganglion cells implies a neural influence on the functional activity of the paraganglia. Some paraganglia in the late fetal and early postnatal specimens contained Timofeew's sensory corpuscles, resembling pacinian corpuscles in their morphology. The central nerve fibre of these corpuscles displayed immunoreactivity for SP, CGRP and NOS, the latter indicating a possible role for NO in afferent transmission from the urinary bladder. In addition, a few corpuscles were penetrated by a noradrenergic nerve fibre immunoreactive for NPY and TH, which may have a modulatory role on the sensory receptor.     

Co-localization of calcitonin gene-related peptide- and substance P-like immunoreactivity in mucosal intra-epithelial nerves in the toad colon.

Mucosal intra-epithelial nerve terminals have been found in the colon of the toad, Bufo marinus. Co-localized in the fibres are calcitonin gene-related peptide (CGRP)-like immunoreactivity (LI) and substance P (SP)-LI. The intraepithelial nerves are likely to be terminals of primary afferent sensory neurones. Fibres with vasoactive intestinal peptide (VIP)-LI or neuropeptide Y (NPY)-LI also supply the mucosa but do not penetrate the epithelium.     

TH-, NPY-, SP-, and CGRP-immunoreactive nerves in interscapular brown adipose tissue of adult rats acclimated at different temperatures: an immunohistochemical study

Interscapular brown adipose tissue (IBAT), a site of nonshivering thermogenesis in mammals, is neurally controlled. The co-existence of sympathetic and peptidergic innervation has been demonstrated in different brown adipose depots. We studied the morphological profile of IBAT innervation and tested by immunohistochemical methods whether cold and warm stimulation are accompanied by modifications in the density of parenchymal noradrenergic nerve fibers. We also studied the immunoreactivity of afferent fibers—which contain calcitonin gene-related peptide (CGRP) and substance P (SP)<197>in different functional conditions. IBAT was obtained from adult rats (6 weeks old) acclimated at different temperatures (4°, 20°, and 28°C). Tissue activity was evaluated by studying the immunolocalization of uncoupling protein (UCP-1), a specific marker of brown adipose tissue. Noradrenergic and peptidergic innervation were seen to arise from morphologically different nerves. Fibers staining for tyrosine hydroxylase (TH) were thin, unmyelinated hilar nerves, and CGRP- and SP-positive fibers were in thick nerves containing both myelinated and unmyelinated fibers. Under cold stimulation, noradrenergic neurons produce greater amounts of TH, and their axons branch, resulting in increased parenchymal nerve fibers density. Neuropeptide Y (NPY) probably co-localizes with TH in noradrenergic neurons, but only in the perivascular nerve fiber network. The parenchymal distribution of NPY to interlobular arterioles and capillaries suggests that this peptide must have other functions besides that of innervating arteriovenous anastomoses, as hypothesized by other researchers. The different distribution of CGRP and SP suggests the existence of different sensory neuronal populations. The detection of CGRP at the parenchymal level is in line with the hypothesis of a trophic action of this peptide.     

Distribution and origin of nerve fibers in the rat temporomandibular joint capsule

 The distribution and origin of nerve fibers containing neuropeptides and NOS projecting to the temporomandibular joint capsule (TMJ) of the rat were studied by retrograde tracing in combination with immunocytochemistry. Numerous nerve fibers were seen in the TMJ as revealed by the neuronal marker protein gene product 9.5. Nerve fibers containing neuropeptide Y (NPY), vasoactive intestinal peptide (VIP), pituitary adenylate cyclase activating peptide (PACAP), substance P (SP), calcitonin gene-related peptide (CGRP), and nitric oxide synthase (NOS) were seen in the synovial membrane, the joint capsule and entering the articular disc. Injection of the retrograde tracer True Blue (TB) into the TMJ resulted in the appearance of numerous labeled nerve cell bodies in the trigeminal and superior cervical ganglia, and moderate numbers in the nodose, the otic, the sphenopalatine, the stellate and the dorsal root ganglia at levels C2–C5. Most of the TB-labeled cell bodies in the superior cervical and stellate ganglia contained NPY. In the trigeminal ganglion, numerous TB labeled cell bodies contained CGRP and a minor population stored SP, a few cell bodies were seen to store NOS or PACAP. In the sphenopalatine and otic ganglia, TB labeled cell bodies contained NOS or VIP. In the nodose ganglion, labeled cell bodies contained CGRP; other labeled cell bodies harbored NOS. In the cervical dorsal root ganglia, the majority of the labeled cell bodies stored CGRP and smaller populations stored SP and PACAP. Thus, the innervation of the TMJ is complex and many different ganglia are involved. Accepted: 13 October 1997