Distribution of enteric nerve cell bodies and axons showing immunoreactivity for vasoactive intestinal polypeptide in the guinea-pig intestine

Immunoreactivity for vasoactive intestinal polypeptide has been localized in neurons in the guinea-pig ileum, colon and stomach. In the ileum, 2.5% of the nerve cell bodies of the myenteric plexus and 45% of those of the submucous plexus showed vasoactive intestinal polypeptide-like immunoreactivity. Varicose axons containing vasoactive intestinal polypeptide ramified amongst the nerve cell bodies of both plexuses and in some cases formed rings of varicosities around non-reactive nerve cells. Axons were traced from the myenteric plexus to the circular muscle and deep muscular plexus. There were numerous positive axons running in fine strands within the circular muscle, parallel to the muscle bundles. Axons containing vasoactive intestinal polypeptide were associated with mucosal blood vessels, but few supplied the vascular network of the submucosa; some immunoreactive axons also contributed to the periglandular plexus of the mucosa. There were no changes in the distribution of axons in the ileum after extrinsic denervation.The results are discussed in relation to the possible functional roles of neurons that contain vasoactive intestinal polypeptide in the intestine: the distribution of such nerve cells in the myenteric plexus and of axons in the circular muscle and sphincters is consistent with this polypeptide being a transmitter of enteric inhibitory neurons; it is also possible that vasoactive intestinal polypeptide is the enteric vasodilator transmitter.     

The plasma concentrations of secretin and vasoactive intestinal polypeptide (VIP) after long-term,strenuous exercise

Summary Twelve subjects (aged 21–38 years) participated in a 90-km cross-country ski race lasting 4.45–6.50 h. In order to investigate a metabolic role for secretin and vasoactive intestinal polypeptide (VIP) during long-term strenuous exercise, seven of the subjects were given an oral hypertonic glucose solution while the others had no nutrient intake immediately after the race. The plasma concentrations of secretin and VIP were greatly increased immediately after the race, and the levels were not normalized within 140 min, though there was a significant decrease after 80 min with rest. The post-race plasma levels of secretin and VIP showed a rapid and very marked decrease 15 min after ingestion of glucose, and the plasma levels had returned to control levels 60 min and 15 min respectively after glucose loading. The results indicate some metabolic function for secretin and VIP during long-term physical exercise.     

An electrophysiological study of the projections of putative sensory neurons within the myenteric plexus of the guinea pig ileum

Conduction of action potentials in the processes of AH (afterhyperpolarizing) neurons has been examined in the myenteric plexus of the guinea-pig small intestine. AH neurons are a morphologically distinct class of myenteric neurons in which the action potentials are followed by long lasting afterhyperpolarizations and which usually lack fast synaptic inputs. These neurons have large smooth cell bodies and several long processes. We have used electrophysiological methods, combined with intracellular injection of the fluorescent dye 5(6)-carboxyfluorescein, to examine the directions of projection and lengths of axons of AH neurons. AH neurons of the myenteric plexus projected circumferentially in both directions from the cell soma for electrophysiologically determined average distances of 0.74 ± 0.05 mm. Thus, the neurons span about 1.5 mm of the circumference of the intestine. About one quarter of the AH neurons had one, or rarely two, processes that ran anally after initially projecting circumferentially. All processes conducted action potentials, with average conduction velocities of 0.23 ± 0.02 ms⁻¹.     

Alterations in the distributory pattern of vasoactive intestinal polypeptide immunoreactivity in the ischemic intestines of dogs

Alterations in the distribution of vasoactive intestinal polypeptide (VIP) in normal and ischemic small intestines of dogs were studied by using conventional transmission electron microscope, and immunohistochemistry for light and electron microscopy. At the light microscopic level, immunoreactivity was evident in the intestinal ganglionic cells of control segments. At the electron microscopic level using a pre-embedding method, the entire cytoplasm of the ganglionic cells in the control segments was filled with VIP immunoreactive products, while the post-embedding experiment showed positive reactions only within the VIP granules and Golgi vesicles. After 30 min of ischemia, immunoreactivity was greatly decreased in the ganglionic cells and a large amount of VIP immunoreactive product appeared in the striated border of epithelial cells and in nerve fibers of the subepithelial layer. These results suggest that intestinal ischemia might lead to the release of VIP, which seems to bind to the microvillus membrane of epithelial cells. The relationship between the changes in VIP distribution and its protecting mechanisms of ischemic damage is discussed.     

Distribution of vasoactive intestinal polypeptide in the rat and mouse brain.

The distribution of cell bodies and nerve fibers that combine with antisera to vasoactive intestinal polypeptide (VIP) was studied by immunohistochemistry in combination with radioimmunoassay in the brain of rat and mouse. The highest concentrations (60pmol/g wet wt) of immuno-reactive VIP were found in the cerebral cortex and in certain limbic structures, whereas the concentrations in the basal ganglia, thalamus, lower brain stem, cerebellum and spinal cord were low (<15pmol/g). VIP-immunoreactive cell bodies were found mainly in the cerebral cortex and the limbic system, with the great majority of them in neo- and allocortical areas. In the neocortex the VIP-containing cell bodies were found in layers II-V in all areas. The cells were fusiform or stellate shaped, resembling intracortical and corticocortical association neurones. In the pyriform and entorhinal cortex the cell bodies were located mainly in layer II. In the hippocampal complex VIP-containing cell bodies occurred in both the subiculum, areas CA1 and CA3 and the dentate gyrus. Most of the cells had the appearance of interneurones, some of them probably being identical with basket cells. Of subcortical areas, the amygdala had the largest number of VIP-containing cell bodies; they were numerous in all amygdaloid nuclei except in the central nucleus. Non-cortical areas where there were cell bodies containing VIP included the anterior olfactory nuclei, the bed nucleus of stria terminalis, lateral septum, suprachiasmatic nucleus, superior colliculus, and the mesencephalic periaqueductal gray.VIP-immunoreactive fibres had a distribution which on the whole paralleled that of the cell bodies, suggesting that many of the VIP-containing cells project locally. VIP-containing fibres were numerous in the following areas: the entire neocortex, the pyrifom cortex, the entorhinal cortex, the hippocampal complex, the amygdala (the central nucleus in particular), the anterior olfactory nuclei, the nucleus accumbens, ventral pallidum, bed nucleus of stria terminalis, suprachiasmatic nucleus, medial preoptic nucleus, median eminence, lateral geniculate body, pretectum, superior colliculus, periaqueductal gray, and the lateral parabrachial nucleus. Only few, scattered fibres were seen in other parts of the brain stem, in the striatum, thalamus and spinal cord. The cerebellum was devoid of VIP-containing fibres. VIP-containing neurones seem to form predominantly local projections. In addition, some VIP-containing neurones probably also form long projections, such as descending and transcallosal projections from the cortical cells, and projections from the amygdala to preoptic, hypothalamic and basal forebrain areas.The characteristic telencephalic distribution of the neurones that contain VIP suggests a role for this peptide in cortical and limbic functions.     

Different effects of visual deprivation on vasoactive intestinal polypeptide (VIP)-containing cells in the retinas of juvenile and adult rats

Vasoactive intestinal polypeptide (VIP) is a 28 amino acid peptide that has been shown to be present in a distinct subset of retinal amacrine cells (VIP+ cells). Previous work has suggested that the expression of retinal VIP might depend on the lighting conditions prevailing. Reasoning that a careful analysis of the exact conditions of this interaction and its time course might offer clues to the functional role of retinal VIP, we performed a quantitative immunohistochemical analysis of the effects of visual deprivation on the retinal VIP+ system in adult and in visually inexperienced neonatal rats. In adult rats visual deprivation caused a marked suppression of VIP-like immunoreactivity (IR) in both somata and processes of VIP+ cells which increased linearly over time, reducing the number of VIP+ cells to about 30% of the control value after approximately 50 days of deprivation. The observed changes were specific for VIP and were independent of the form of deprivation used (monocular lid suture versus keeping rats in constant darkness). However, the effects of visual deprivation were dependent on the developmental state of the rats, since juvenile rats without previous visual experience exhibited a decrease in VIP+ cells and fibers which was much smaller and occurred significantly slower than in adult rats. The suppression of VIP-like IR was completely reversible in both juvenile and adult rats when previously deprived rats were returned to a normal visual environment.     

Changes of cerebral vasoactive intestinal polypeptide-and somatostatin-like immunoreactivity induced by noise and whole-body vibration in the rat

To clarify the role of vasoactive intestinal polypeptide (VIP) and somatostatin, somatropin-release inhibiting factor, (SRIF) neurons in the response to organisms to noise or whole-body vibration stress, VIP and SRIF-like immunoreactivity were determined in various regions of the rat brain following exposure for 90 min to noise (broad band, 102 dB) or whole-body vibration (20 Hz, 4.0 g). Both noise and whole-body vibration significantly increased VIP-like immunoreactivity in the amygdala. A significant reduction of VIP like immunoreactivity in the hippocampus was induced only by whole-body vibration. On the other hand SRIF-like immunoreactivity was decreased significantly in the hypothalamus and increased significantly in the amygdala by noise and whole-body vibration, respectively. The present findings would seem to indicate that the amygdalofugal VIP neural system is involved in regulating hypothalamic and pituitary hormone secretions in non-specific reactions to stress. Responses of hippocampal VIP and the amygdalofugal SRIF to whole-body vibration stress are assumed to be activated as specific reactions to the stress.     

Co-localization of neuropeptide Y, vasoactive intestinal polypeptide and dynorphin in non-noradrenergic axons of the guinea pig uterine artery

Two major populations of perivascular axons containing immunoreactivity to neuropeptide Y (NPY) have been revealed in the main uterine artery of the guinea pig by immunohistochemical procedures which allow the simultaneous visualization of two antigens. One population contained immunoreactivity to dopamine-beta-hydroxylase (D beta H) and was presumably noradrenergic. The other main population of axons with NPY-like immunoreactivity (NPY-LI) did not have D beta H-like immunoreactivity (D beta H-LI) and was presumably non-noradrenergic. These non-noradrenergic axons also contained immunoreactivity to vasoactive intestinal polypeptide (VIP) and dynorphin (DYN). Indeed, nearly all axons with VIP-LI also contained NPY-LI and DYN-like immunoreactivity (DYN-LI). NPY constricted the uterine artery perfused in vitro, whilst VIP dilated uterine arteries preconstricted with noradrenaline or NPY. Thus, we have evidence for the coexistence of a vasoconstrictor peptide and a vasodilator peptide in the same non-noradrenergic perivascular axons, which also contain an opioid peptide, dynorphin.     

Subcellular fractionation of cat submandibular gland: Comparative studies on the distribution of acetylcholine and vasoactive intestinal polypeptide (VIP)

In previous studies, evidence has been obtained for the presence of vasoactive intestinal polypeptide (VIP) in presumptive cholinergic neurons innervating the exocrine glands in the cat. In the present study, an attempt was made to define the storage sites of these two compounds in the cat submandibular gland using subcellular fractionation techniques. Particulate VIP was preferentially found in dense fractions (0.78–0.97 M sucrose) of a density gradient. Particle-bound acetylcholine showed a bimodal distribution in the gradient with relative enrichment in a dense fraction (0.87 M sucrose) but mainly in lighter fractions (0.28–0.44 M sucrose). More than 50% of all acetylcholine was recovered in a soluble form. For comparison, noradrenaline was also analysed and found in paniculate form in fractions ranging between 0.44–0.78 M sucrose. The electron microscopic analysis revealed presence of i.a. small clear vesicles in the lighter fractions, whereas the fractions rich in VIP contained i.a. many larger vesicles, sometimes with an electron-dense core. In agreement with earlier ultrastructural immunocytochemical studies on intact tissue, the present results support the view that VIP is present in large dense-cored vesicles. Most of the particulate acetylcholine seems to be localized in small clear vesicles, although a small proportion could be associated with large vesicles. Whether acetylcholine and VIP coexist in such large vesicles or whether separate populations of large vesicles exist, remains to be elucidated.     

P物质、血管活性肠肽和乙酰胆碱能神经在大鼠肠道内的分布及其关系

应用乙酰胆碱酯酶(AChE)组织化学和PAP免疫组织化学方法,比较观察P物质(SP)、血管活性肠肽(VIP)和AChE三种阳性神经元在大鼠十二指肠、空肠、回肠、结肠和直肠内的分布特征及其相互关系。结果显示:SP、VIP、AChE阳性神经神经元和纤维均分布于肠壁各层,从十二指肠、空肠到回肠逐渐增多,但从结肠到直肠则逐渐减少;AChE阳性神经元或纤维在肠壁各层最丰富,其中VIP以粘膜层和粘膜下神经丛较丰富,SP以肠肌丛较丰富;三者的分布密度为AChE>VIP>SP。AChE、SP和VIP阳性神经元胞体及神经纤维在不同肠段的分布密度有明显差异(P<0.05),提示可能与不同肠段肠动力调节功能有关。     

Localization of vasoactive intestinal polypeptide in penile erectile tissue and in the major pelvic ganglion of the rat

Vasoactive intestinal polypeptide was localized by immunocytochemical techniques in the major pelvic ganglion and penile erectile tissue of the rat. Vasoactive intestinal polypeptide fibers were concentrated in penile crura with the density of innervation decreasing distally. The helicine arteries were very densely innervated while fewer fibers surrounded the deep artery of the penis. Intrinsic smooth muscle of the cavernous bodies received a moderate supply of vasoactive intestinal polypeptide immunoreactive fibers. Dorsal vascular structures, including the deep dorsal vein were innervated by vasoactive intestinal polypeptide fibers. Vasoactive intestinal polypeptide immunoreactive cell bodies were found in the major pelvic ganglion, concentrated on one end of the ganglion. Rectrograde studies with a dye injected into the penile crura indicated that neurons in major pelvic ganglion projected to the penis. Combined dye and immunofluorescent studies showed that all the dye-labeled neurons were immunoreactive for vasoactive intestinal polypeptide.

It is concluded that all vascular beds in the penis of the rat are innervated by vasoactive intestinal polypeptide fibers and that the extent of the innervation is related to the occurrence of smooth muscle. Neurons in the major pelvic ganglion probably are the main source of vasoactive intestinal polypeptide fibers to the penis.     

Synaptology and sources of vasoactive intestinal polypeptide and substance p containing axons of the cat celiac ganglion. An experimental electron microscopic immunohistochemical study

We have examined the possible origin of vasoactive intestinal polypeptide (VIP) and substance P-containing axons and the synapses formed by these axons in the celiac ganglion of the cat, by means of ultrastructural immunohistochemistry combined with various surgical lesions.Axons containing VIP as well as those immunopositive for substance P, formed axo-dendritic and axosomatic synapses with principal ganglion cells. After transaction of the superior mesenteric nerve numerous degenerated axonal profiles could be found in the celiac ganglion. Some of these contained VIP immunoreactivity. By contrast in alternate sections stained with substance P antibody only intact axons were labelled.Bilateral vagotomy resulted in the appearance of degenerating axonal profiles in the celiac ganglion, some of which could be stained with substance P antiserum but not with VIP antiserum. Following removal of dorsal root ganglia (Th₆-Th₁₂) from both sides, a large number of degenerated axons were found, many of which were immunopositive for substance P but not for VIP.We conclude that the peripheral input to the celiac ganglion contains VIP fibers which form synapses with principal ganglion cells. Substance P-containing fibers reach the celiac ganglion via the vagal nerve as well as from the dorsal root ganglia, and they form synapses with the principal ganglion cells. It is proposed that the VIP and the substance P-containing axons are involved in two different peripheral reflex loops.     

Possible modulatory role of dynorphin on the excitation by neurotensin on the guinea pig myenteric plexus

Dynorphin_1–13 antagonized in a concentration-dependent fashion the contractile effect of neurotensin on the isolated preparation of the guinea pig ileum myenteric plexus. The inhibitory action of dynorphin was reduced in the presence of naloxone, indicating the opioid nature of this interaction. Atropine also reduced the excitatory component of the neurotensin-induced contractile response; the joint application of atropine plus dynorphin did not cause additional inhibition of the contractile effect of neurotensin.     

Distribution of vasoactive intestinal polypeptide-, calcitonin gene-related peptide-, somatostatin- and neurofilament-immunoreactivities in sympathetic ganglia of human fetuses and premature neonates

The distribution patterns of vasoactive intestinal polypeptide (VIP), calcitonin gene-related peptide (CGRP), somatostatin (SOM) and neurofilament (NF) immunoreactivities (IR) were studied in the stellate ganglia of human fetuses and neonates at 24-26 weeks gestation. Sizeable populations with some quantitative variations of VIP-, CGRP- and SOM immunoreactive nerve cells were detected in all ganglia studied. In marked contrast, neurofilament expression was down-regulated. The upregulation of VIP, CGRP and SOM expression suggested their inductor effect on growth and differentiation neurons as well as on the development of their neurotransmitter properties. The main neuropeptides-inducing factor of sympathetic ganglia in human prenatal ontogenesis may be considered as a relative hypoxia.     

Perivascular nerves with immunoreactivity to vasoactive intestinal polypeptide in cephalic arteries of the cat: distribution, possible origins and functional implications

The distribution of nerves containing vasoactive intestinal polypeptide(VIP)-immunoreactive material was examined in the cephalic arteries and cranial nerves of cats using an indirect immunofluorescence procedure on whole mounts. Perivascular VIP-immunoreactive nerves were widely distributed in arteries and arterioles supplying glands, muscles and mucous membranes of the face. Within the cerebral circulation, perivascular VIP-immunoreactive nerves were most abundant in the circle of Willis and the proximal portions of the major cerebral arteries and their proximal branches supplying the rostral brainstem and ventral areas of the cerebral cortex. Nerves containing VIP-immunoreactive material were absent from distal portions of arteries supplying the posterior brainstem, cerebellum and dorsal cerebral cortex. Cerebral perivascular VIP-immunoreactive nerves had extracerebral origins probably from VIP-immunoreactive perikarya within microganglia in the cavernous plexus and external rete. Extracerebral perivascular VIP-immunoreactive nerves probably arose from VIP-immunoreactive perikarya in microganglia associated with the tympanic plexus, chorda tympani, lingual nerve and Vidian nerve as well as from cells in the otic, sphenopalatine, submandibular and sublingual ganglia. Therefore, it seems likely that each major segment of the cephalic circulation is supplied by local VIP-immunoreactive neurons. If the VIP-immunoreactive nerves cause vasodilation, they are well placed to allow redistribution of arterial blood flow within the head. During heat stress, neurogenic vasodilation of the appropriate beds would permit efficient cooling of cerebral blood, particularly that supplying the rostral brainstem and surrounding areas of the cerebral cortex.     

Effects of age and streptozotocin-induced diabetes on contents and effects of substance P and vasoactive intestinal polypeptide in the lower urinary tract of the rat

Andersson , P. O., Fahrenkrug , J., Malmgren , A. & Uvelius , B. 1992. Effects of age and streptozotocin-induced diabetes on contents and effects of substance P and vasoactive intestinal polypeptide in the lower urinary tract of the rat. Acta Physiol Scand 144 , 361–368. Received 29 January 1 991 , accepted 11 October 1991. ISSN 0001–6772. Departments of Physiology, Clinical Pharmacology and Urology, University of Lund, Lund, Sweden and Department of Clinical Chemistry, Bispebjerg Hospital, Copenhagen, Denmark. The urinary bladder and urethral content of substance P and vasoactive intestinal polypeptide and the in vitro effects of the peptides on the bladder were studied at 6 weeks and 6 months of streptozotocin-induced diabetes in the rat. The results were compared with those obtained in age-matched control animals. Both short-term and long-term streptozotocin treatment induced a clearcut increase in bladder weight. Bladder substance P content was increased in both groups of diabetic animals but substance P concentration was similar in control and diabetic animals. Vasoactive intestinal polypeptide content was slightly higher in diabetic animals than in controls but vasoactive intestinal polypeptide concentration was significantly lower in the bladders from both short-term and long-term diabetic animals. The bladder contractile response to substance P was similar in all groups of animals and vasoactive intestinal polypeptide was found to be devoid of contractile or relaxatory effects in the rat bladder. No change in urethral weight was seen with diabetes. There were no clear-cut changes in the urethral contents or concentrations of substance P and vasoactive intestinal polypeptide. The study also enabled comparisons between younger (3 months) and older (9 months) rats. This comparison showed a decrease in the concentrations and contents of substance P and vasoactive intestinal polypeptide between young and older rats. The changes were seen in both the bladder and the urethra and were similar in diabetic and normal animals.     

The GDNF-induced neurite outgrowth and neuronal survival in dissociated myenteric plexus cultures of the rat small intestine decreases postnatally

 Glial cell-line-derived neurotrophic factor (GDNF), a member of the transforming growth-factor- (TGF-) β-family, is an essential factor for the development of the enteric nervous system (ENS) during embryogenesis. In the present study, the effects of GDNF on postnatal ENS development were investigated using cultures of myenteric plexus from the small intestine of newborn albino rats of different developmental phases (P1, P7, P14). Myenteric plexus was dissociated and cultivated as mixed cultures of enteric neurons and glial cells. After seeding, the cultures were kept for 24 h or 7 days in serum-free medium containing various doses (1, 10, 100 ng/ml) of GDNF. The effect of the neurotrophic factor was evaluated using parameters such as cell size, neuronal survival, or neurite elongation. While neither glial-cell nor neuronal size was influenced by GDNF, there was an observable effect upon neuronal survival and neurite elongation. The cultures treated with GDNF displayed increased neurite outgrowth. The promoting effect was dose- and age-dependent, decreasing clearly during the early postnatal period. Already after 24 h, neuronal survival was increased in P1 and P7, but not in P14 cultures. In long-term cultures, a marked tendency to form cell aggregates and dense fiber networks was observed when treated with GDNF. These observations suggest that GDNF plays an important role not only in pre-, but also in postnatal development of the enteric nervous system. Received: 29 May 1998 / Accepted: 10 December 1998