Vasoactive Intestinal Polypeptide Immunoreactive Nerve Research Articles

Morphological evidence of local reflex arc in the rat’s tongue

Lingual components of the autonomic nervous system are considered to be the most rostral portion of the enteric nervous system. Therefore our aim was to study the intrinsic nerve cell bodies and synapses using immunohisto-, immunocytochemical methods. Several small groups of ganglia with cell bodies immunoreactive (IR) for vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) and substance P (SP) were observed just below the gustatory epithelium. A few somatostatin and galanin IR nerve cell bodies were also found. Many IR cell bodies were also demonstrated in the glands and next to blood vessels. Some of these cell bodies were multipolar and some of them were small neurons with an ovoid shape having only one process. Cell bodies positive for calcitonin gene-related peptide (CGRP) were detected neither in the superficial nor in the deep portion. Electronmicroscopical analysis demonstrated different IR nerve fibres having axo-somatic and axo-dendritic synapses with other immunonegative cells. In a few cases VIP IR nerve processes were found to synaptize with other VIP positive nerve cell bodies. These results support the existance of intralingual reflex in the tongue, where the ganglia might have an integrative role of the different neuropeptide containing nerve fibres.

Distribution and possible origin of neuropeptide-containing nerve elements in the mammalian liver

The intrahepatic distribution of nerve fibres is highly species dependent, therefore we searched for a species where the innervation pattern is similar to that of the human liver. Livers of rats, cats, guinea pigs and humans were used. The different nerve elements were identified by ABC immunohistochemistry and analysed semiquantitatively. Large numbers of neuropeptide Y (NPY) and dopamine-beta-hydroxylase immunoreactive (IR) nerve fibres were observed in the human and guinea pig liver, and they were in close contact with portal triads, central veins and ran parallel with liver sinuses. A few substance P, somatostatin and vasoactive intestinal polypeptide IR nerve fibres were also detected intralobularly, while galanin nerve fibres were only observed around portal triads. In the rat liver only a few NPY-positive nerve fibres were found, exclusively in portal tracts. Some nerve cell bodies (IR for NPY and somatostatin) were also found in the liver of guinea pigs, young cats and humans, therefore some of the nerve terminals might originate from these intrinsic ganglia. It can be concluded that the innervation pattern of the guinea pig liver shows the highest similarity to that of the human liver.

Re-innervation pattern of the ‘neovagina’ created from the bladder flap in patients with Mayer-Rokitanski-Kistner-Hauser syndrome: An immunochemical study

Background. Cystovaginoplasty (CVP) is a method of vaginal reconstruction in women with Mayer-Rokitansky-Kistner-Hauser Syndrome (MRKHS). The neo-vagina allows normal sexual intercourses, but after CVP, the sexual life of patients with MRKHS does not differ significantly from normal females. Therefore, we decided to elucidate the pattern of sensory re-innervation of the bladder flap used for the surgery.Methods. Biopsies were taken from vaginal vestibule and urinary bladder during the CVP and 1 year later in four patients with MRKHS. The following neurotransmitters were studied calcitonin gene-related peptide (CGRP), galanin (GAL), vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase-activating peptide (PACAP).Results. CGRP and PACAP nerve fibres were sparse under the urothelium and in submucosal layer of the neovagina, they were more numerous around blood vessels and in the vicinity of smooth muscles. This was similar to the pattern observed in the urinary bladder. VIP- and GAL-positive nerve fibres were most numerous in the submucosa around blood vessels and in smooth muscle bundles of neovagina. They were distinctly less numerous beneath the epithelium. This innervation pattern mimicked that seen in normal vagina and in vaginal vestibule of patients with MRKHS.Conclusions. Our findings demonstrate considerable nervous system plasticity in the bladder flap. Distribution of presumably sensory CGRP and PACAP immunoreactive nerve fibers was similar to the pattern observed within the intact bladder wall, and VIP or GAL immunoreactive nerve fibers (vasomotor functions) were distributed in a manner similar to that observed in the intact vaginal wall.

Development of Vasoactive Intestinal Polypeptide-Immunoreactive Nerves in the Major Cerebral Arteries of the Quail Anterior Circulation

Development of cerebral perivascular nerves immunoreactive for vasoactive intestinal polypeptide (VIP) was investigated in the Japanese quails, using immunohistochemistry and quantitative analysis. VIP-immunoreactive (VIP-IR) nerves supplying the anterior circulation appeared on the cerebral carotid artery (CCA) at embryonic day 10 and on the cerebroethmoidal artery (CEA) after hatching. Nerves from the CCA increased greatly in number and spread progressively during successive embryonic stages, while those from the CEA were sparse all through the post-hatching stages, mostly remained limited to this vessel wall. The distribution of VIP-IR nerves to the respective major arteries of the anterior circulation from the two vascular routes was basically similar among post-hatching day (P) 15, P20, P30 and P50. Likewise, no clear statistical difference was observed with regard to the nerve density of the corresponding arteries in the four age groups. These findings suggest that VIP-IR innervation of the quail anterior circulation usually attains its mature pattern at the third week after hatching.

Different Patterns of Vasoactive Intestinal Polypeptide (VIP)-Immunoreactive and Acetylcholinesterase (AChE)-Positive Innervation in the Internal Carotid Artery and Cerebral Arterial Tree of the Quail

The innervation pattern of vasoactive intestinal polypeptide-immunoreactive (VIP-IR) nerves in the quail internal carotid artery (ICA) and cerebral arterial tree was investigated and compared with that of acetylcholinesterase-positive (AChE-P) nerves. The supply of VIP-IR nerves to the two arterial systems was distinctly richer than that of AChE-P nerves. It was focused mainly on the walls from the distal ICA to the caudal half of the anterior ramus (AR) through the cerebral carotid artery (CCA). Indeed, double staining clearly showed that numerous VIP+/AChE-axons were distributed over these arterial regions where VIP+/AChE+ or AChE+/VIP- axons were sporadic or often lacking. The finding that nerve bundles accompanying the ICA within the carotid canal contained abundant VIP+/AChE- nerve cells suggests that cerebrovascular VIP-IR nerves in the quail have their major source at these neurons and enter the cranial cavity through the CCA. Another significant finding was that a small number of nerve cells, which were mostly stained for AChE alone and occasionally for VIP alone or both, occurred in the major arteries located more rostral than the middle AR. Thus, the quail cerebral arterial tree, at least the rostral segment of the anterior circulation, is multiply innervated by these three distinct categories of the extracranial and intracranial VIP-IR and AChE-P neurons.

Separate Development of Nitric Oxide Synthase- and Vasoactive Intestinal Polypeptide-Immunoreactive Nerves Arising from the Vertebral Artery in the Rat

Development of nitric oxide synthase (NOS)-and vasoactive intestinal polypeptide (VIP)-immunoreactive (-IR) nerves supplying the basilar and vertebral arteries (BA and VA) was investigated in White Wistar rats, using double immunohistochemistry. NOS-IR and VIP-IR nerves via the anterior circulation (AC), which mostly expressed NO(+)/VIP(+), extended to the BA during the second postnatal week, and usually reached as far as the rostral two third of the BA on PND 20. NOS-IR nerves were completely lack in the cBA and the VA on PND10, and often absent from these arterial regions even at PND 20. Nevertheless, a small number of VIP(+)/NOS(-) nerves were localized in the walls from the caudal BA (cBA) to the VA on PND 5. On PND 20, they frequently met with the descending NOS-IR and VIP-IR nerves via the AC around the lower portion of the middle BA. Fiber bundles containing NOS(+)/VIP(+) axons were first visualized on the caudal VA at PND 30 and observed frequently at PND 80, with a distinct increase in number of NOS-IR and VIP-IR nerves supplying the cBA and the VA. Thus, NOS-IR nerves coming from the VA develop through its own characteristic sequence that lags markedly behind the time of appearance for VIP-IR nerves from the same vascular route and for NOS-IR and VIP-IR nerves via the AC.

Changes in mouse sudomotor function and sweat gland innervation with ageing

Age-related changes in sudomotor neuroeffector function have been evaluated in mice aged 2 (young), 6, 12 (adult) and 18 (old) months. We evaluated sudomotor function by determining the number of sweat glands reactive to pilocarpine and the sweat output per gland on the plantar surface of the hindpaws with the impression mould technique. Protein gene product 9.5 (PGP) and vasoactive intestinal polypeptide (VIP) were immunohistochemically localised in footpads. A marked decrease (44%) in sweat output per gland was observed in old mice as well as a slight (17%), not significant decline in the number of secreting sweat glands. The sudomotor innervation, expressed as the area of sweat gland occupied by VIP and PGP immunoreactive nerve profiles, showed an initial increase from 2 to 6 months and a significant decline (35%) in 18- vs. 6-month-old mice. These results indicate that, in contrast to the number of secreting sweat glands, sweat output per gland does not reach the maximum in adult mouse until 6 months old and that sweating decreases in aged mice mainly due to a decline of sweat output per gland and to a lesser extent to a decrease in number of secreting glands. A reduction of sweat glands size in aged mice was also found, suggesting that the diminished sweat gland responsiveness with ageing may be attributed to sweat gland atrophy as well as to loss of innervation.

Large veins in the Atlantic cod (Gadus morhua) and the rainbow trout (Oncorhynchus mykiss) are innervated by neuropeptide-containing nerves.

Previous studies have shown that fish veins are reactive to several hormones known to exist in the fish circulation. Besides this humoral control, another possible means of active regulation of venous return is by autonomic nervous control of venous tone. This study therefore investigated the presence of a perivascular innervation of major veins in the Atlantic cod (Gadus morhua) and the rainbow trout (Oncorhynchus mykiss) using immunohistochemical methods. Histological staining was employed to investigate the smooth muscle distribution in the vessel walls. Vasoactive intestinal polypeptide-immunoreactive nerve fibers were found to be widespread in the venous system of G. morhua and O. mykiss, while pituitary adenylate cyclase-activating polypeptide-immunoreactive fibers were demonstrated in the duct of Cuvier of both species. Fibers containing neurokinin A and/or substance P were found in the duct of Cuvier and the posterior cardinal vein of both species and in the hepatic portal vein of O. mykiss. Calcitonin-gene related peptide-immunoreactive fibers were present in the duct of Cuvier of both species and in the hepatic portal vein of O. mykiss. Galanin-immunoreactive fibers were found in the duct of Cuvier in O. mykiss and in the hepatic portal vein of both species. Co-existence of neuropeptides in the perivascular nerve fibers was investigated by double labelling. Vasoactive intestinal polypeptide/pituitary adenylate cyclase-activating polypeptide-immunoreactive fibers were found in both species. Vasoactive intestinal polypeptide/galanin-immunoreactive fibers and vasoactive intestinal polypeptide/calcitoningene related peptide-immunoreactive fibers were found in G. morhua but not in O. mykiss. This study gives further evidence for an active venoregulation by autonomic nerves in teleost fish.

Comparative study on the appearance of various bioactive peptides in foregut derivates during the ontogenesis

Bioactive peptides have an important multifunctional role in the gastrointestinal tract. In the present study we have investigated the dynamism of the appearance of PACAP (pituitary adenylate cyclase activating polypeptide), VIP (vasoactive intestinal polypeptide), gastrin, and secretin immunoreactivities in human foregut derivates during the ontogenesis using an immunohistochemical approach. None of these peptides were observed in the foregut derivates of an 8-week-old embryo. VIP immunoreactive nerve fibers appeared by the 11th week in the smooth muscle layers of the stomach. No other peptide immunoreactivities were observed of this stage. In 18- and 20-week old fetuses PACAP, secretin, and gastrin immunoreactive cells appeared in the developing glands of the stomach. In the duodenum gastrin immunoreactivity was present in the Lieberkühn's glands and secretin immunoreactive cells were seen between the surface epithelial cells. In the pancreas secretin immunoreactivity was found in the Langerhans islets; however, PACAP immunreactivity was observed in the exocrine portion. The distribution of VIP fibers did not change during the fetal life and it was similar to the adult pattern. According to our results the appearance of PACAP, secretin, and gastrin in the developing glands suggests their role in the proliferation and differentiation of the epithelial derivates.

Immunohistochemical localization of the VIP1 receptor (VPAC1R) in rat cerebral blood vessels: relation to PACAP and VIP containing nerves.

The two structurally related peptides, vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase activating polypeptide (PACAP), are present in cerebral vascular nerve fibers. Biologic actions of VIP are exerted through two receptors, VPAC1 and VPAC2, having similar binding affinity for both VIP and PACAP. In the current study, the authors have developed a specific antibody against the rVPAC1 receptor to examine the localization of rVPAC1 immunoreactivity in cerebral arteries and arterioles of the rat by immunohistochemistry using fluorescence confocal microscopy. Specificity of the antiserum was ensured by immunoblotting and immunocytochemistry of cells transfected with cDNA encoding the different PACAP-VIP receptor subtypes. The rVPAC1 receptor immunoreactivity was localized to the plasmalemma of circularly orientated smooth muscle cells on superficial cerebral arteries and arterioles taken from the basal surface of the brain. By double immunostaining VIP immunoreactive nerve fibers and, to a lesser extent, those containing PACAP were shown to have intimate contact with the receptor protein. Vasoactive intestinal polypeptide and PACAP containing cerebrovascular nerve fibers were found in separate nerve populations with different distribution pattern and density. In brain sections processes of cortical VIP-, but not PACAP-, containing neurons seemed to innervate the rVPAC1 receptor of pial arterioles on the brain surface. The current findings provide the neuroanatomical substrate for a role of VIP and maybe PACAP in the regulation of cerebral blood flow.

Regional difference in the distribution of vasoactive intestinal polypeptide-immunoreactive nerve fibres along the uterus and between myometrial muscle layers in the rat.

To investigate a possible regional variation of the vasoactive intestinal polypeptide innervation in the uterus of the cyclic rat, the distribution of vasoactive intestinal polypeptide-containing nerve fibres from the cervix to the oviduct end of the uterine horns was studied using immunohistochemistry. Immunoreactive nerve fibres were most concentrated in the cervix, where they formed a dense plexus in association with the musculature and surrounding blood vessels. In the uterus, a clear regional distribution of the vasoactive intestinal polypeptide innervation was observed. Numerous vascular and non-vascular immunoreactive nerve fibres were present in the lower part of the uterine horns, whereas they were sparse in the median region and absent at the oviduct end. Moreover, non-vascular peptide innervation was mostly concentrated in the circular layer of the myometrium and also occurred in the endometrium. Only a very few immunoreactive nerve fibres were present in the longitudinal muscle layer. No change in the peptide innervation pattern was observed during the different stages of the sexual cycle. The marked regional distribution of the peptide innervation in the rat uterus suggests that the regulatory effects of the peptide occur mainly in the lower part of the organ and principally affect the circular muscle layer in the myometrium.

Vasoactive intestinal polypeptide immunoreactive nerves in the gill arch of teleost fish, Carassius auratus L.

The localization of vasoactive intestinal polypeptide (VIP)-immunoreactive (ir) nerve cell bodies and fibers has been studied in the gill arches of goldfish (Carassius auratus, L.) using the peroxidase-antiperoxidase (PAP) immunohistochemical method. It was found that VIP-ir nerve cell bodies are localized in connective tissue on the oral side of the gill arch; these cells were present as single cells, in couples or as small clusters. Moreover, a dense network of VIP-ir fibers was observed beneath the lining epithelium of the raker cushion. The possible involvement of this peptide in mucus secretion in the gill arches of teleost is discussed.

Inhibition of nitroxidergic nerve function by neurogenic acetylcholine in monkey cerebral arteries.

1. Modification by endogenous or exogenous acetylcholine and vasoactive intestinal polypeptide (VIP) of vasodilatation mediated by nitric oxide (NO) released from nitroxidergic nerves was studied in isolated monkey cerebral arteries. In arterial strips denuded of endothelium, transmural electrical stimulation (2-20 Hz) produced relaxations that were abolished by tetrodotoxin. 2. The relaxation response was attenuated by acetylcholine, and the attenuation was reversed by atropine. Attenuation was also observed with AF-DX 116, an antagonist of the muscarinic acetylcholine receptor subtype, M2. NO-induced relaxation was not affected by acetylcholine. Neurogenic relaxation was also inhibited by physostigmine and potentiated by atropine. 3. VIP in concentrations that elicited slight relaxation did not alter the response to nerve stimulation. In the strips showing tachyphylaxis to VIP, the neurogenic response was not inhibited. 4. Histochemical studies of whole-mount preparations revealed nerve fibres with NO synthase and VIP immunoreactivity, and also acetylcholinesterase, suggesting the presence of perivascular nitroxidergic, VIPergic and cholinergic innervation. 5. It is concluded that the actions of nitroxidergic nerve fibres on the monkey cerebral artery are inhibited by nerve-released acetylcholine acting on prejunctional muscarinic receptors, possibly of the M2 subtype. Despite the presence of VIP immunoreactive nerve fibres and the ability of exogenous VIP to relax the artery, there is no evidence supporting either a prejunctional modulation of nitroxidergic nerve function by VIP or a role for VIP as a vasodilatory neurotransmitter.

Distribution of vasoactive intestinal polypeptide immunoreactive nerve fiber in the rat nasal mucosa

The distribution of vasoactive intestinal polypeptide (VIP)-immimoreactive nerve fiber of the respiratory nasal mucosa of the rat was examined by use of immunohistochemistory with light, laser confocal scanning and electron microscopy. On light microscopy, it was observed that VIP immunoreactive nerve fibers distributed under and within the epithelium in addition to around the blood vessels and glands. The laser confocal scanning microscope revealed the network of VIP-immunoreactive nerve fibers at glands, but it was denser around glands than within glands. The VIP-immunoreactive nerve fibers formed fine varicosities and was distributed below the middle portion of the epithelium. Some fibers were observed as if they were surrounding goblet cells. Electron microscopy showed that the VIP-immunoreactive nerve fibers within the epithelium were terminated as free nerve endings. Some of them were in close contact with goblet cells.

Selective damage to sensorimotor perivascular nerves in the mesenteric vessels of diabetic rats.

The perivascular innervation of the superior mesenteric artery and vein was examined using immunohistochemical and immunoassay techniques in rats 8 weeks after induction of diabetes with streptozotocin (STZ). Increased density of innervation and fluorescence intensity was noted for substance P- and calcitonin gene-related peptide-immunoreactive nerves in the diabetic vessels. A slight increase in the density of vasoactive intestinal polypeptide-immunoreactive nerve fibers innervating the mesenteric artery was also noted. However, there was no change in the density of neuropeptide Y- and dopamine beta-hydroxylase-immunoreactive nerve fibers, although the fluorescence intensity of neuropeptide Y-immunoreactive nerve fibers was reduced in diabetic rat vessels. Immunoassays showed that the levels of substance P- and calcitonin gene-related peptide were increased > 10-fold in the diabetic mesenteric vein, while levels of neuropeptide Y and vasoactive intestinal polypeptide were unchanged. In summary, there is a marked increase in nerve fibers containing sensory neuropeptides in mesenteric vessels of STZ-induced diabetic rats, which, in view of the reported impaired sensorimotor function in these vessels, is likely to reflect a neuropathic change.

Neurotransmitters for the Canine Inferior Pharyngeal Constrictor Muscle

The inferior pharyngeal constrictor muscle plays an important role at the pharyngeal phase of deglutition and is anatomically composed of the thyropharyngeal muscle and cricopharyngeal muscle. In this study we investigated the distribution pattern of neuropeptidergic and catecholaminergic nerve fibers in the thyropharyngeal muscle and cricopharyngeal muscle of seven puppies by immunohistochemistry. Some of the calcitonin gene‐related peptide–, substance P–, vasoactive intestinal polypeptide–, and tyrosine hydroxylase–immunoreactive nerve fibers were found to lie parallel to the muscle fibers in both the thyropharyngeal muscle and cricopharyngeal muscle. Nerve fibers with immunoreactivity to all substances examined were found to be associated with blood vessels in both the thyropharyngeal muscle and cricopharyngeal muscle, and the number of calcitonin gene‐related peptide, neuropeptide Y, and tyrosine hydroxylase nerve fibers was higher than the number of substance P, vasoactive intestinal polypeptide, and galanin nerve fibers. Motor end plate‐like structures with calcitonin gene‐related peptide immunoreactivity were found in both the thyropharyngeal muscle and cricopharyngeal muscle. These structures in the cricopharyngeal muscle were clearly less than those in the thyropharyngeal muscle. Some clusters of neurons were detected only in the cricopharyngeal muscle of all dogs examined. Substance P–, vasoactive intestinal polypeptide–, galanin–, and neuropeptide Y–immunoreactive neurons were found in this ganglion, and the vasoactive intestinal polypeptide–immunoreactive neurons were the most abundant. Abundant calcitonin gene‐related peptide– and vasoactive intestinal polypeptide–immunoreactive nerve fibers, and some substance P– and galanin‐immunoreactive nerve fibers were distributed in the ganglion. A few neuropeptide Y– and tyrosine hydroxylase‐immunoreactive nerve fibers were also found in the ganglion. These findings suggest the complex participation of neuropeptides in the innervation of the inferior constrictor muscle.

Neuropeptide Y- and vasoactive intestinal polypeptide-like immunoreactivity in adjuvant arthritis: effects of capsaicin treatment.

The occurrence of the neuropeptides vasoactive intestinal polypeptide (VIP) and neuropeptide Y (NPY) in ankle joints and dorsal root ganglia (L2-L6) was analyzed in normal and arthritic Lewis rats. In addition the effect of capsaicin pretreatment was investigated. The study included 92 rats consisting of 4 groups, 23 rats in each; normal rats, normal rats given capsaicin, arthritic rats and arthritic rats pretreated with capsaicin. The localization of the neuropeptides was assessed by immunohistochemistry and the tissue concentrations were determined by radioimmunoassay (RIA). In the arthritic rats, there was a slight increase in NPY immunoreactive nerve fibres in the ankle joint synovium and bone marrow, as compared to normal rats. Notably, there was an intense fluorescence and significant increase (p < 0.01, 41%) in the number of NPY-positive megakaryocytes in the tibial bone marrow of arthritic rats. RIA showed that the concentration of NPY-like immunoreactivity (LI) was increased by 50% in the ankle joint. Pretreatment with capsaicin did not affect the increased level of NPY-LI in the ankle joint of arthritic rats. The concentration of NPY-LI in the dorsal root ganglia was not altered in arthritic rats, nor was it affected by the capsaicin treatment. No NPY immunoreactive cells could be detected in the dorsal root ganglia. The number of VIP immunoreactive nerve fibres observed in ankle joints of arthritic and normal rats did not differ. However, RIA measurements showed an 11% increase in the VIP concentration in arthritic rats, which was unaffected by capsaicin treatment. In dorsal root ganglia, RIA disclosed a 21% increase in VIP-LI, although no VIP-positive cells could be detected. Capsaicin treatment did not affect the increased concentration of VIP-LI in the dorsal root ganglia.

Quantitative immunohistochemical assessment of bovine myocardial innervation before and after cryosurgical cardiac denervation.

The aim was to determine the relative distribution and possible origins of peptide containing nerves in the bovine heart before and after functional extrinsic denervation established by cryosurgery. A quantitative immunohistochemical technique was used. In the intact heart, myocardial nerve fibres and fascicles displaying immunoreactivity for the general neural marker protein gene product 9.5 had an atrial to ventricular gradient in density. The right atrium was the most densely innervated region and a major proportion of the total myocardial innervation, visualised by protein gene product 9.5 immunoreactive nerves, showed neuropeptide tyrosine (45%) and tyrosine hydroxylase (20%) immunofluorescence staining, while nerves immunoreactive for vasoactive intestinal polypeptide and calcitonin gene related peptide formed relatively minor subpopulations (representing less than 2% and 0.5%, respectively, of the total fluorescent myocardial innervation). Following cryoablation there was a significant reduction in the percentage fluorescent area of protein gene product 9.5 immunoreactive nerves throughout the heart, of greater than 90% of the control values. There were highly significant reductions in the percentage fluorescent area of nerves showing immunoreactivity to neuropeptide tyrosine, tyrosine hydroxylase, and calcitonin gene related peptide, to 1.01, 0.92, and 0.05%, respectively, of the intact myocardial innervation. The distribution of vasoactive intestinal polypeptide immunoreactive nerves was more variable and displayed an equivocal response to cardiac cryoablation. The majority of nerves showing immunoreactivity to neuropeptide tyrosine, tyrosine hydroxylase, and calcitonin gene related peptide are of extrinsic origin, while vasoactive intestinal polypeptide immunoreactive nerves may have intrinsic as well as extrinsic origins. The distribution and apparent origins of immunohistochemically defined nerves in the bovine heart are similar to those observed in the human heart which suggests that the calf may be an appropriate model for comparative studies.

Neuropeptide Y, tyrosine hydroxylase and vasoactive intestinal polypeptide-immunoreactive nerve fibers in the vertebral bodies, discs, dura mater, and spinal ligaments of the rat lumbar spine.

The occurrence of autonomic nerves in the lumbar spine of rats was investigated by immunohistochemical technique. Both peptidergic nerves, represented by immunoreactivity to neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), and noradrenergic nerves as reflected by tyrosine hydroxylase (TH) immunoreactivity, were identified. NPY- and TH-positive nerves were predominantly found in the blood vessels walls. They occurred in the bone and periosteum of the vertebral body, disc, dura mater, and in the spinal ligaments. They were particularly numerous along the growth plate and in the outer layers of the intervertebral discs. VIP-positive fibers were predominantly nonvascular. They occurred in all tissues analyzed, but were most abundant in the vertebral bone marrow and periosteum. The VIP-immunoreactive fibers in the outer fibrous layers of the disc and the spinal ligaments were occasionally observed in blood vessel walls. No immunoreactivity could be detected in the nucleus pulposus. In the dura mater, NPY-, TH- and VIP-positive fibers were found both in the ventral and dorsal portion. In view of the vasoconstrictive properties of both NPY and noradrenaline, it may be assumed that the abundance of NPY- and TH-immunoreactive nerves fibers in blood vessel walls reflects a vasoregulatory activity. The predominance of nonvascular VIP-positive fibers in the vertebral bone marrow and periosteum may represent an involvement in local bone physiology.

Guanethidine sympathectomy of mature rats leads to increases in calcitonin gene-related peptide and vasoactive intestinal polypeptide-containing nerves

Changes in the innervation of the heart (right atrium), mesenteric blood vessels, vas deferens and superior cervical ganglia have been examined following long-term sympathectomy of the mature rat. Patterns of innervation were investigated by histochemical and immunohistochemical techniques, while levels of noradrenaline and neuropeptides were measured by neurochemical assays. Large doses of guanethidine (80mg/kg) were given daily for four weeks to 12–14 week-old male rats which were killed at 18–20 weeks of age. Catecholamine-containing nerves were severely depleted or absent in all tissues, together with a reduction in noradrenaline content. Neuropeptide Y levels were depleted by 97% in vas deferens, 78% in mesenteric vein and 50% in right atrium and superior cervical ganglion. Increases in levels of calcitonin gene-related peptide were seen in the mesenteric vein (up seven-fold), superior cervical ganglia (up 11-fold) and vas deferens (prostatic portion up three-fold), which were also evident by assessment of immunolabelling of nerve fibres. Calcitonin gene-related peptide levels were not increased in the right atrium. In addition, an increase in vasoactive intestinal polypeptide-immunoreactive nerve fibre density was seen in the mesenteric artery and vas deferens, although no significant differences were observed in assays of vasoactive intestinal peptide levels in any tissue. No changes were seen in the innervation of any of the tissues by substance P-immunoreactive nerve fibres either by immunohistochemical or immunochemical assay assessment. This study indicates that there are selective changes in the mature nervous system in response to the loss of sympathetic nerves. Differences between these changes and the response of the developing nervous system to long-term sympathectomy are discussed.