Presence Of Vasoactive Intestinal Polypeptide Research Articles

Enteroendocrine cells couple nutrient sensing to nutrient absorption by regulating ion transport

The ability to absorb ingested nutrients is an essential function of all metazoans and utilizes a wide array of nutrient transporters found on the absorptive enterocytes of the small intestine. A unique population of patients has previously been identified with severe congenital malabsorptive diarrhea upon ingestion of any enteral nutrition. The intestines of these patients are macroscopically normal, but lack enteroendocrine cells (EECs), suggesting an essential role for this rare population of nutrient-sensing cells in regulating macronutrient absorption. Here, we use human and mouse models of EEC deficiency to identify an unappreciated role for the EEC hormone peptide YY in regulating ion-coupled absorption of glucose and dipeptides. We find that peptide YY is required in the small intestine to maintain normal electrophysiology in the presence of vasoactive intestinal polypeptide, a potent stimulator of ion secretion classically produced by enteric neurons. Administration of peptide YY to EEC-deficient mice restores normal electrophysiology, improves glucose and peptide absorption, diminishes diarrhea and rescues postnatal survival. These data suggest that peptide YY is a key regulator of macronutrient absorption in the small intestine and may be a viable therapeutic option to treat patients with electrolyte imbalance and nutrient malabsorption.

Concentration of vasoactive intestinal polypeptide, neuropeptide Y and calcitonin gene related peptide in neoplastic tissue of the human large intestine

Aim of the study: The purpose of this study was to investigate the presence of vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) and calci tonin gene related peptide (CGRP) neu ropeptides in neoplastic tissue of the large intestine. The results obtained were compared to an unchanged part of the intestine originating from the same patient. Material and methods: The tissue sam ples were taken from the postoperative part of the intestine, resected during an operation due to treatment of colon cancer. Tissues were taken from 15 patients. Concentrations of neu ropeptides were determined by ELISA tests, and results were calculated per 1 g of fresh tissue. The data obtained were statistically analysed. Results: In tissues of the neoplastic tumour of the sigmoid colon and rec tum the concentration of VIP and NPY was determined (28.9 ng/g and 8.6 ng/g, respectively). The observed values were statistically significantly lower than those observed in corresponding tissues of unchanged intestinal seg ments (83.9 ng/g for VIP and 19.5 ng/g for NPY). Surprisingly, amounts of CGRP were very similar in both neoplastic and intact tissues (13.3 ng/g and 13.4 ng/g respectively). Conclusions: Immunoenzymatic inves tigations allow one to establish the con centration of neuropeptides associated with the enteric nervous system in can cer tissues of the large intestine. Vari ous concentrations of these neuropep tides were observed in particular patients of both compared groups. The performed statistical analysis did not reveal associations between degree of cancer infiltration within the colonic wall and the level of presence of inves tigated neuropeptides in either cancer or control tissues. The fact of presence of these neuropeptides in cancer tissues confirmed by this study should be underlined at the same time.

ACh- and VIP-induced vasorelaxation in rabbit facial artery after carotid artery occlusion

Objectives The influence of carotid artery occlusion (10, 30 and 60 min) on regulatory mechanisms implicated in the vasorelaxant responses of isolated glandular branch of rabbit facial artery to acetylcholine (ACh) and vasoactive intestinal polypeptide (VIP) was examined. Design In organ bath studies with arterial rings precontracted with phenylephrine (1 μM), before and after carotid artery occlusion, changes in isometric tension were recorded. Results Endothelium-dependent vasorelaxation by ACh and endothelium-independent vasorelaxation by VIP were significantly reduced, started from 30 and 10 min of carotid occlusion, respectively. Inhibitory effect of indomethacin on ACh vasorelaxation was enhanced whilst effect of N G-nitro- l-arginine reduced, started from 30 min of carotid occlusion. Sodium nitroprusside-induced vasorelaxation was not changed after carotid occlusion. Inhibition of VIP vasorelaxation by l-N ω-nitroarginine-2,4- l-diaminobutyric-amide, was reduced, started from 30 min of carotid occlusion. Forskolin enhanced VIP-induced vasorelaxation in control rings but this effect was reduced started from 30 min of occlusion. In the presence of VIP, vasorelaxant effect of ACh was increased; the increase was reduced, started from 10 min of carotid occlusion. Conclusions The present investigation provides evidence for the decreased responsiveness to both, ACh-endothelium-dependent and VIP-endothelium-independent vasorelaxation in rabbit facial artery after carotid occlusion. In addition, the data suggest that ischaemia alters contribution of endothelial nitric oxide (eNO) and prostaglandin to ACh, and vascular smooth muscle's cAMP and neuronal NO to VIP vasorelaxant effects.

Pituitary adenylate cyclase-activating polypeptide, vasoactive intestinal polypeptide and their receptors: distribution and involvement in the secretion of Podarcis sicula adrenal gland

Vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) are regulatory neuropeptides of the hypothalamus-hypophyseal-adrenal axis, acting via the common receptors VPAC(1) and VPAC(2) and the selective PACAP receptor PAC(1). In the adrenal glands of the Italian wall lizard, Podarcis sicula, the presence of VIP in chromaffin cells, and the VIP-stimulated release of catecholamine and aldosterone in vivo, was previously shown. To examine the localization of both peptides and receptors and their mRNAs in the adrenal gland of P. sicula, immunohistochemistry and in situ hybridization were performed: PACAP and its mRNA were detected in chromaffin cells, VPAC(1) was found associated with steroidogenic tissue, VPAC(2) and PAC(1) with chromaffin tissue. Using 'far western blot' technique, we showed the presence of specific binding sites for VIP/PACAP in the adrenal glands of the lizard. The effects of both VIP and PACAP on the adrenal cells of the lizard were examined in vitro in adrenal cell co-cultures: both VIP and PACAP enhanced catecholamine, corticosterone and aldosterone release from adrenal cell co-culture in a time- and dose-dependent manner. The catecholamine release was inhibited by PAC(1) antagonist and in VPAC(2) immunoneutralized adrenal cells. The effects of VIP and PACAP on aldosterone secretion were counteracted by VPAC(1) antagonist administration in vitro. Corticosterone secretion elicited by VIP was not blocked by VPAC(1) antagonist, while the PACAP-induced release of corticosterone was blocked by the antagonist. Overall, our investigations indicate that these neuropeptides of the secretin superfamily can act not only as neurotransmitters but also as autocrine and paracrine regulators on chromaffin and cortical cells, being important mediators of the non-cholinergic system in the lizard adrenal gland.

NANC inhibitory neuromuscular transmission in the hamster distal colon

The neurotransmitter(s) that generate the inhibitory junctional potential (IJP) in the circular muscle of hamster distal colon and their mechanisms have not been elucidated. The aim of the present study, therefore, was to determine the contributing roles of the non-adrenergic, non-cholinergic (NANC) inhibitory transmitter(s) including nitric oxide (NO), adenosine 5′-triphosphate (ATP) and vasoactive intestinal polypeptide (VIP) in the generation of IJP in the hamster distal colon. For this purpose, the effects of the corresponding blockers of these putative NANC inhibitory mediators have been investigated using microelectrode technique. Intracellular membrane potential recordings were made from smooth muscle cells at 35 °C in Tyrode's solution that contained atropine (0.5 μM), guanethidine (3 μM) and nifedipine (0.5 μM). Single electrical stimuli (0.5 ms, 50 V) as well as trains of two and five pulses (20 Hz at the same duration and voltage) elicited NANC IJP consisted of initial fast (IJP-F) followed by a slow hyperpolarization (IJP-S). The response had been abolished by tetrodotoxin (TTX, 0.3 μM). The nitric oxide synthase (NOS) inhibitor, N G-nitro-L-arginine methyl ester (L-NAME; 200 μM) blocked IJP-S but enhanced IJP-F. The later had been blocked with suramin, a universal P2 receptor antagonist, or with CBF3GA, a P2Y receptor antagonist at dose-dependent fashions. The IJP-F had been markedly inhibited by desensitization of P2Y receptor with its putative agonist 2-methylthio-ATP (2-meSATP, 50 μM for 30 min). IJP-F was sensitive to the P2Y1 receptor specific antagonist A3P5PS (10 μM) and to the G-protein inhibitor, pertussis toxin (PTX, 400 ng/ml for 2 h) as well as to the small and intermediate Ca 2+ sensitive K + channels blocker, apamin (0.3 μM). IJP-S was blocked by the guanylate cyclase (GC) inhibitor, 1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one (ODQ, 10 μM) and was partially sensitive to apamin. Exogenously applied ATP (100 μM–1 mM) produced typical hyperpolarization that was blocked by suramin, CBF3GA and 2-meSATP desensitization; while exogenously applied NO (3–10 μM) produced slowly developing hyperpolarization that was not blocked by L-NAME but ODQ. In the presence of both purinergic and nitrergic inhibitors, stimulation using a train of eight pulses at 25 Hz evoked a small slow hyperpolarization that was sensitive to the VIP antagonist (VIP 6–28, 1 μM). Exogenous application of VIP (1–10 μM) produced similar response that was not evident in the presence of VIP 6–28. These data indicate that NANC IJP that is generated in the circular muscle cells of hamster distal colon is mediated by ATP and NO via P2Y1/P2Y2 receptor and GC-dependent pathways, respectively. A masked role for VIP is also indicated.

Dendritic cells get VIP treatment

Comment on Fujita et al, page [3656][1], Gonzalez-Rey et al, page [3632][2], and Chorny et al, page [3787][3] Three manuscripts in this issue of Blood describe induction of immune tolerance by dendritic cells (DCs) generated in vitro in the presence of vasoactive intestinal polypeptide (VIP) or IL-

Presence and co-localization of vasoactive intestinal polypeptide with neuronal nitric oxide synthase in cells and nerve fibers within guinea pig intrinsic cardiac ganglia and cardiac tissue

The presence of vasoactive intestinal polypeptide (VIP) has been analyzed in fibers and neurons within the guinea pig intrinsic cardiac ganglia and in fibers innervating cardiac tissues. In whole-mount preparations, VIP-immunoreactive (IR) fibers were present in about 70% of the cardiac ganglia. VIP was co-localized with neuronal nitric oxide synthase (nNOS) in fibers innervating the intrinsic ganglia but was not present in fibers immunoreactive for pituitary adenylate cyclase-activating polypeptide, choline acetyltransferase (ChAT), tyrosine hydroxylase, or substance P. A small number of the intrinsic ChAT-IR cardiac ganglia neurons (approximately 3%) exhibited VIP immunoreactivity. These few VIP-IR cardiac neurons also exhibited nNOS immunoreactivity. After explant culture for 72 h, the intraganglionic VIP-IR fibers degenerated, indicating that they were axons of neurons located outside the heart. In cardiac tissue sections, VIP-IR fibers were present primarily in the atria and in perivascular connective tissue, with the overall abundance being low. VIP-IR fibers were notably sparse in the sinus node and conducting system and generally absent in the ventricular myocardium. Virtually all VIP-IR fibers in tissue sections exhibited immunoreactivity to nNOS. A few VIP-IR fibers, primarily those located within the atrial myocardium, were immunoreactive for both nNOS and ChAT indicating they were derived from intrinsic cardiac neurons. We suggest that, in the guinea pig, the majority of intraganglionic and cardiac tissue VIP-IR fibers originate outside of the heart. These extrinsic VIP-IR fibers are also immunoreactive for nNOS and therefore most likely are a component of the afferent fibers derived from the vagal sensory ganglia.

The increase in prolactin-secreting cells in incubating chicken hens can be mimicked by extended treatment of pituitary cells in vitro with vasoactive intestinal polypeptide (VIP)

Incubation of eggs by birds and lactation in mammals are regulated by pituitary prolactin (PRL) and associated with an increase in pituitary PRL-producing cells or lactotrophs. However, the mechanisms controlling this increase in lactotroph numbers are not known. PRL secretion in birds is regulated by vasoactive intestinal polypeptide (VIP). This study was designed to determine whether VIP treatment could modulate lactotroph abundance in culture. Anterior pituitary cells were isolated from laying Japanese White Silkie hens and cultured for 2 or 6 days in the absence or presence of VIP. PRL-secreting cells were identified by reverse hemolytic plaque assay. Treatment with VIP for 6 days substantially increased the abundance of PRL-secreting cells from 47.5% under basal conditions to 70.6% of all pituitary cells following VIP stimulation. However, 2-day VIP treatment had no effect. Furthermore, the extent to which the hens were allowed to accumulate eggs in a clutch prior to isolation of the pituitaries did not affect the lactotroph response to VIP in vitro . These findings indicate that chronic VIP stimulation may be responsible for the increased abundance of lactotrophs found in the pituitary glands of incubating hens.

Distribution of vasoactive intestinal polypeptide in the rat heart: effect of guanethidine and capsaicin

Vasoactive intestinal polypeptide (VIP) is believed to coexist with acetylcholine in postganglionic parasympathetic neurones. However, the presence of VIP in extrinsic nerves and/or other types of intrinsic cardiac neurones has not been excluded. The aim of our study was to examine the distribution and origin of VIP-ergic innervation in the rat heart atria using immunocytochemistry and radioimmunoassay (RIA) combined with two types of denervation: sympathectomy, which was produced by guanethidine treatment and sensory denervation achieved by capsaicin administration. In whole-mount preparations of the intact atria, VIP-immunoreactive (IR) nerve fibres and ganglionic cells were found, the latter being much more numerous in the left atria (LA) than in the right ones. Some of VIP-IR nerve fibres forming bundles appeared to be extrinsic in origin. VIP-IR concentrations determined by RIA in the intact rats were significantly higher in the LA than in the right ones (p < 0.01). However, no changes in VIP-IR levels were found in either atrium after both guanethidine and capsaicin treatment protocols, thus indicating that VIP-immunoreactivity is not associated with either sympathetic or sensory innervation. In conclusion, the ganglionated plexus of the rat atria may comprise at least 3 different neuronal populations expressing VIP-positivity: 1. extrinsic preganglionic parasympathetic fibres, 2. intrinsic postganglionic parasympathetic neurones and 3. intrinsic local circuit neurones that do not express a cholinergic phenotype.

Comparative distribution of VIP in the central nervous system of various species measured by a new radioimmunoassay.

Vasoactive intestinal polypeptide (VIP) occurs in high concentrations throughout the gut and the nervous system. The presence of VIP has been shown in a number of species, mainly by immunohistochemistry. The aim of the present study was to develop a new, highly specific VIP radioimmunoassay to investigate the distribution of VIP in the central nervous system of various vertebrate and invertebrate species. Different areas of the brain and spinal cord were removed from rats, chickens, turtles, frogs and fishes. The cerebral ganglia and the ventral ganglionic chain were investigated in the earthworm. The tissue samples were processed for VIP radioimmunoassay. Our results show that the antiserum used in the radioimmunoassay turned to be C-terminal specific, without significant affinity to other members of the VIP peptide family. Detection limit of the assay was 0.1 fmol/ml. Highest concentrations were found in the turtle diencephalon, followed by other brain areas in the turtle and rat. All other brain areas in the examined species contained significant levels of VIP. Immunoreactivity was also shown in the cerebral and ventral ganglia of the earthworm. In summary, our results show comparative quantitative distribution in representative species of the phylogenetic line, using the same experimental conditions.

The influence of GnRH, oxytocin and vasoactive intestinal peptide on the secretion of β-endorphin and production of cAMP and cGMP by porcine pituitary cells in vitro

The objective of this study was to determine whether gonadotrophin-releasing hormone (GnRH), oxytocin (OT) and vasoactive intestinal polypeptide (VIP) modulate β-endorphin-like immunoreactivity (β-END-LI) secretion by dispersed anterior pituitary cells of pigs and in vivo priming with steroid hormones, estradiol benzoate (EB) and progesterone (P 4), influences the cell reactivity to peptide hormones tested. Additionally, the aim of this research was to examine the involvement of cyclic nucleotides (cAMP and cGMP) in transduction of signals induced by GnRH, OT and VIP in porcine pituitary cells. Pituitaries were collected from ovariectomized (OVX) gilts that were divided into four experimental groups. Animals of group 1 (OVX) received 1 ml corn oil (placebo)/100 kg body weight (b.w.), group 2 (OVX+EB I) and group 3 (OVX+EB II) were treated with EB at the dose 2.5 mg/100 kg b.w., 30–36 and 60–66 h before slaughter, respectively. Animals of group 4 (OVX+P 4) were injected with P 4 at the dose 120 mg/100 kg b.w. for 5 subsequent days before slaughter. Anterior pituitaries were dispersed with trypsin and then pituitary cells were cultured (10 6 per well) in McCoy’s 5A medium containing horse serum (10%) and fetal calf serum (2.5%) for 3 days at 37 °C under an atmosphere of 95% air and 5% CO 2. Subsequently, plates were rinsed with fresh McCoy’s 5A medium and pituitary cells were treated with one of the following agents: GnRH (100 ng/ml), OT (10 −6 M) or VIP (10 −7 M) and incubated for 3.5 h at 37 °C. GnRH did not affect β-END-LI secretion by pituitary cells of OVX (group 1) and OVX+P 4 (group 4) gilts. When the pituitary cells were incubated in the presence of OT and VIP, significant increases were observed. After priming of OVX gilts with EB, 30–36 h before slaughter (group 2), we noted a significant increase in β-END-LI release from pituitary cells only in the presence of VIP. Pituitary cells from gilts treated with EB, 60–66 h before slaughter (group 3), produced markedly elevated amounts of β-END-LI after GnRH, OT or VIP addition. GnRH markedly stimulated cGMP release from cultured pituitary cells in all experimental groups and significantly increased cAMP production by the cells from OVX, OVX+EB II and OVX+P 4 animals. The addition of OT enhanced both cAMP and cGMP output in all experimental groups of pigs. VIP stimulated cAMP release from pituitary cells derived from OVX, OVX+EB I and OVX+EB II animals. cGMP output was markedly elevated under the influence of VIP from pituitary cells of OVX, OVX+EB II and OVX+P 4 gilts. In conclusion, our results suggest that GnRH, OT and VIP can modulate β-endorphin release from porcine pituitary cells and imply the involvement of cAMP and cGMP in transduction of signals induced by studied peptides in the cells.

The relationships of nicotinamide adenine dinucleotide phosphate-d to nitric oxide synthase, vasoactive intestinal polypeptide, galanin and pituitary adenylate activating polypeptide in pigeon gut neurons

The distribution of nicotinamide adenine dinucleotide phosphate (NADPH)-d neurons and their relationship with nitric oxide synthase (NOS), vasoactive intestinal polypeptide (VIP), pituitary adenylate activating polypeptide (PACAP) and galanin (Gal) were examined in the gastrointestinal (GI) tract of the pigeon Columbia livia. NADPH-d-histochemistry, indirect immunofluorescence and confocal analysis were applied to cryosections. Western blot analysis was also applied on pigeon gut. NADPH-d neurons were found throughout the pigeon GI tract and they were evident in the myenteric, circular muscle and submucous plexuses. Positive varicose nerve fibres were also distributed within the longitudinal muscle layers and in the lamina propria of the mucosa. The stomach was the segment richest in positivities. The copresence VIP/Gal/NOS as well as PACAP/VIP were revealed in some NADPH-d-neurons. We suppose that the nitrergic nerve population of the pigeon GI tract belong to the muscle motility regulation as an inhibitory descending nerve pathway. Moreover the presence of VIP, Gal and PACAP in some NADPH-d-containing neurons enhances the inhibitory actions of these neurotransmitters whereas PACAP and Gal role is actually unknown.

Immunohistochemical Detection of Vasoactive Intestinal Polypeptide (VIP) and the VIP Receptor in the Rat Inner Ear

Vasoactive intestinal polypeptide (VIP) is a 28 amino acid peptide that was originally isolated from porcine duodenum. The presence of VIP has been demonstrated in the central nervous system and peripheral tissues. However, there have been few reports on VIP as a neurotransmitter, especially in the inner ear. To investigate the function of VIP in the rat inner ear, we examined the expression of VIP and the VIP receptor by immunohistochemistry. Using the anti-VIP and VIP receptor antibodies, scattered fibres in the cochlear nerve trunk demonstrating VIP-like immunoreactivity were found, and the spiral ganglion cells demonstrated ring-shaped VIP-like immunoreactivity. Immunoreactivity for the VIP receptor was predominantly found in the spiral ganglion cells. Our results suggest that VIP may play an important role as a possible neurotransmitter not only in the local control of cochlear blood flow, but also in the auditory system.

An immunohistochemical screening of neurochemical markers in fungiform papillae and taste buds of the anterior rat tongue

The occurrence and distribution of several neurochemical markers were investigated. Numerous nerve fibres were shown, using antibodies to protein gene product (PGP) 9.5, neurone-specific enolase, calcitonin gene-related peptide (CGRP), substance P, neurokinin A or protein S-100. The presence of vasoactive intestinal polypeptide (VIP), peptide histidine isoleucine amide (PHI), neuropeptide tyrosine, dopamine-β-hydroxylase (DBH), cholecystokinin/gastrin, glutamate and galanin was more scarce. Nerve fibres containing these above-mentioned markers were found at several locations, i.e. in the epithelium, connective tissue, and around blood vessels. In the taste buds, numerous PGP 9.5-, neurone-specific enolase-, CGRP-, substance P-, neurokinin A- and protein S-100-containing structures were found, but few VIP and galanin ones. No immunoreactivity was found with antibodies against somatostatin, bombesin, enkephalin or dynorphin. These findings extend knowledge about the general as well as the neurochemical messenger-based innervation of rat fungiform papillae, forming a firm basis for future functional investigations of normal, experimental and also clinical materials.

Presynaptic modulation by VIP, secretin and isoproterenol of somatostatin release from enriched enteric synaptosomes: role of cAMP

The release of somatostatin-like immunoreactivity was studied in isolated enteric synaptosomes. A significant release of somatostatin-like immunoreactivity was observed in the presence of vasoactive intestinal polypeptide (VIP) (10 −6 M: 53.0 ± 12.4 pg/mg, basal: 14.3 ± 1.7 pg/mg, n = 5, P < 0.05), secretin (10 −6 M: 56.1 ± 3.8 pg/mg, basal: 25.8 ± 1.6 pg/mg, n = 6, P < 0.01) and isoproterenol (10 −5 M: 54.0 ± 13.4 pg/mg, basal: 20.0 ± 3.4 pg/mg, n = 8, P < 0.05). Forskolin, an unspecific activator of the adenylate cyclase, caused a significant release of somatostatin-like immunoreactivity (10 −6 M: 57.3 ± 13.2 pg/mg, basal: 30.0 ± 5.8 pg/mg, n = 13, P < 0.01) which was further augmented in the presence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX 10 −4 M) (77.0 ± 17.8 pg/mg, n = 13, P < 0.01). 3-Isobutyl-1-methylxanthine and N 6,2′- O-dibutyryladenosine-3′5′-cyclic monophosphate mimicked the effect of forskolin and VIP. The release of somatostatin was paralleled by an increase of cAMP immunoreactivity in the presence of VIP (10 −6 M: 37.1 ± 9.4 pmol/mg, basal: 19.8 ± 4.2 pmol/mg, n = 10, P < 0.05), isoproterenol (10 −5 M: 42.4 ± 9.8 pmol/mg, basal: 16.7 ± 2.4 pmol/mg, n = 12, P < 0.01) and forskolin (10 −6 M: 47.1 ± 12.4 pmol/mg, basal: 19.8 ± 4.2 pmol/mg, n = 10, P < 0.01). The effect of nitric oxide (NO) which acts as an inhibitory neurotransmitter in the enteric nervous system was studied. NO is known to activate soluble guanylate cyclase to induce transmitter release. The NO-generating compound sodium nitroprusside and bromoguanosine-3′,5′-cyclic monophosphate (8-Br-cGMP) had no effect on the release of somatostatin-like immunoreactivity. These data demonstrate the stimulatory effect of VIP, secretin and isoproterenol on release of somatostatin-like immunoreactivity from enteric synaptosomes, which is presumably mediated by cAMP-dependent mechanisms. cGMP-dependent mechanisms seem to be of minor relevance.

The significance of vasoactive intestinal polypeptide (VIP) in immunomodulation

Evidence for VIP influences on immune function comes from studies demonstrating VIP-ir nerves in lymphoid organs in intimate anatomical association with elements of the immune system, the presence of high-affinity receptors for VIP, and functional studies where VIP influences a variety of immune responses. Anatomical studies that examine the relationship between VIP-containing nerves and subpopulations of immune effector cells provide evidence for potential target cells. Additionally, the presence of VIP in cells of the immune system that also possess VIP receptors implies an autocrine function for VIP. The functional significance of VIP effects on the immune system lies in its ability to help coordinate a complex array of cellular and subcellular events, including events that occur in lymphoid compartments, and in musculature and intramural blood circulation. Clearly, from the work described in this chapter, the modulatory role of VIP in immune regulation is not well understood. The pathways through which VIP can exert an immunoregulatory role are complex and highly sensitive to physiological conditions, emphasizing the importance of in vivo studies. Intracellular events following activation of VIP receptors also are not well elucidated. There is additional evidence to suggest that some of the effects of VIP on cells of the immune system are not mediated through binding of VIP to its receptor. Despite our lack of knowledge regarding VIP immune regulation, the evidence is overwhelming that VIP can interact directly with lymphocytes and accessory cells, resulting in most cases, but not always in cAMP generation within these cells, and a subsequent cascade of intracellular events that alter effector cell function. VIP appears to modulate maturation of specific populations of effector cells, T cell recognition, antibody production, and homing capabilities. These effects of VIP are tissue-specific and are probably dependent on the resident cell populations within the lymphoid tissue and the surrounding microenvironment. Different microenvironments within the same lymphoid tissue may influence the modulatory role of VIP also. Effects of VIP on immune function may result from indirect effects on secretory cells, endothelial cells, and smooth muscle cells in blood vessels, ducts, and respiratory airways. Influences of VIP on immune function also may vary depending on the presence of other signal molecules, such that VIP alone will have no effect on a target cell by itself, but may greatly potentiate or inhibit the effects of other hormones, transmitters, or cytokines. The activational state of target cells may influence VIP receptor expression in these cells, and therefore, may determine whether VIP can influence target cell activity. Several reports described in this chapter also indicate that VIP contained in neural compartments is involved in the pathophysiology of several disease states in the gut and lung. Release of inflammatory mediators by cells of the immune system may destroy VIP-containing nerves in inflammatory bowel disease and in asthma. Loss of VIPergic nerves in these disease states appears to further exacerbate the inflammatory response. These studies indicate that altered VIP concentration can have significant consequences in terms of health and disease. In addition, the protective effects of VIP from tissue damage associated with inflammatory processes described in the lung also may be applicable to other pathological conditions such as rheumatoid arthritis, anaphylaxis, and the swelling and edema seen in the brain following head trauma. While VIP degrades rapidly, synthetic VIP-like drugs may be developed that interact with VIP receptors and have similar protective effects. Synthetic VIP-like agents also may be useful in treating neuroendocrine disorders associated with dysregulation of the hypothalamic—pituitary—adrenal axis, and pituitary release of prolactin.

Ultrastructural study of the presence of vasoactive intestinal polypeptide and serotonin in mucosal nerve fibres and endocrine cells of the intestine of goldfish ( Carassius auratus ) and tilapia ( Oreochromis mossambicus )

In order to establish a possible role of vasoactive intestinal polypeptide (VIP) and serotonin as (neuro)transmitters involved in the regulation of fish intestinal epithelium, we studied the presence of VIP and serotonin at the ultrastructural level in the intestinal mucosa of tilapia and goldfish. A low percentage of varicosities near the basal membrane of the tilapia intestinal epithelium was found to label for VIP or for serotonin, whereas in the goldfish, this percentage was much higher. The varicosities usually contained large granular and small clear vesicles. Immunogold labeling indicated that serotonin and VIP were localized in the large granular vesicles. Unlabeled large granular vesicles and small clear vesicles were usually also present in varicosities with serotonin- or VIP-labeled vesicles. In the goldfish, the serotonin-labeled varicosities were close to the epithelial cells, and direct contacts between serotonin-labeled nerve fibres and epithelial cells could sometimes be visualized. However, synaptic membrane specializations were never observed. In tilapia, the distance between the VIP- or serotonin-labeled varicosities and the epithelial cells was large (more than 2 μm).

Pharmacological approach to the insulin-resistance/polymetabolic syndrome

Incubation of eggs by birds and lactation in mammals are regulated by pituitary prolactin (PRL) and associated with an increase in pituitary PRL-producing cells or lactotrophs. However, the mechanisms controlling this increase in lactotroph numbers are not known. PRL secretion in birds is regulated by vasoactive intestinal polypeptide (VIP). This study was designed to determine whether VIP treatment could modulate lactotroph abundance in culture. Anterior pituitary cells were isolated from laying Japanese White Silkie hens and cultured for 2 or 6 days in the absence or presence of VIP. PRL-secreting cells were identified by reverse hemolytic plaque assay. Treatment with VIP for 6 days substantially increased the abundance of PRL-secreting cells from 47.5% under basal conditions to 70.6% of all pituitary cells following VIP stimulation. However, 2-day VIP treatment had no effect. Furthermore, the extent to which the hens were allowed to accumulate eggs in a clutch prior to isolation of the pituitaries did not affect the lactotroph response to VIP in vitro. These findings indicate that chronic VIP stimulation may be responsible for the increased abundance of lactotrophs found in the pituitary glands of incubating hens.

Vasoactive Intestinal Polypeptide and Thyrotropin-Releasing Hormone Stimulate Newly Synthesized, not Stored, Prolactin*

Experiments were designed to determine whether vasoactive intestinal polypeptide (VIP), reported to stimulate basal PRL secretion, affects PRL processing by lactotrophs. Initially, rat anterior pituitary quarters were incubated for 2 h with [3H]leucine, with and without 10(-5) M VIP, and immunoreactive and immunoprecipitable rPRL were measured during 56 mM KCl perifusion to determine total and 3H-labeled PRL, respectively. Inclusion of VIP increased immunoreactive PRL (P less than 0.05), decreased immunoprecipitable PRL (P less than 0.01), and, therefore, decreased the specific activity of labeled PRL (P less than 0.001). These results suggested an enhanced release of newly synthesized PRL before KCl depolarization, thus decreasing the release of labeled PRL. To discriminate between the two PRL pools, newly synthesized and storage, pituitary quarters were incubated with and without 10(-5) M VIP for 4 h with [14C]leucine, 2 h in cold medium and 2 h with [3H]leucine. Immunoprecipitable PRL was measured during perifusion with 56 mM KCl. Data were depicted as the 3H/14C disintegrations per min ratio of PRL released/3H/14C disintegrations per min of total tissue to account for any differences in tissue labeling. This ratio was greater for tissue labeled in the presence of VIP (P less than 0.002). To determine whether VIP, as a secretagogue, differentiates between the newly synthesized and storage pools, VIP was added after pulse chase, as previously described. No preferential release was observed between the two groups. Finally, using the same [3H]- and [14C]leucine-labeling protocol with and without 10(-5) M VIP, tissue was perifused with medium 199 for 1 h, with 10(-5) M TRH for 30 min, with medium 199 for 30 min, and with 56 mM KCl for 1 h. Inclusion of VIP increased the 3H/14C released/3H/14C total tissue ratio during basal perifusion (P less than 0.04) and TRH exposure (P less than 0.05). Within the control group, the TRH ratio was greater than basal (P less than 0.003). These experiments suggest that newly synthesized PRL is preferentially secreted over stored PRL from tissue incubated with VIP during pulse-chase labeling; however, addition of VIP as a secretagogue did not affect either PRL pool preferentially.

Vasoactive intestinal polypeptide enhances hormone content and insulin release in cultured fetal rat islets.

The effect of porcine vasoactive intestinal polypeptide (VIP) on development of the biphasic insulin release response in cultured fetal rat islets was investigated. Fetal islets, 21.5 days gestational age, were cultured for 7 days in RPMI 1640 culture medium containing either 2.8 or 11.1 mM glucose adn subsequently challenged with 16.7 mM glucose in a perfusion system. Islets were exposed to VIP at a final concentration of 13.2 nM by adding the peptide to the perifusion buffer (acute exposure) or by adding it to the culture medium throughout the culture period (chronic exposure). Islet hormone and DNA contents were also quantitated at the end of the culture period. Acute exposure to VIP resulted in no alterations of the insulin release pattern after culture in the presence of either glucose concentration. However, chronic treatment of islets with 13.2 nM VIP in the presence of 2.8 mM glucose resulted in significant increases in the maximum rate of insulin release during the first phase and the total amount of insulin release during both phases. Similarly, islets cultured in the presence of 11.1 mM glucose and 13.2 nM VIP demonstrated enhanced biphasic insulin release patterns with increased maximum rate and total amount of release during both phases. The presence of VIP and 2.8 mM glucose increased islet glucagon and somatostatin contents, but islet DNA and insulin contents remained unchanged. These findings indicate that VIP plays a significant role in the in vitro development of the biphasic insulin release pattern and may be a factor controlling the maturation of the fetal islet in vivo.