Regulation of G protein mRNA levels by thyroliberin, vasoactive intestinal peptide and somatostatin in prolactin-producing rat pituitary adenoma cells.

We have investigated the regulation of mRNA levels of alpha- and beta-subunits of guanine nucleotide-binding regulatory proteins (G proteins) by peptide hormones in prolactin producing rat pituitary adenoma cells (GH3 cells) in culture. The cells were treated with thyroliberin (1 microM), vasoactive intestinal peptide (1 microM) or somatostatin (10 microM) for 6 to 48 hours. Thyroliberin and vasoactive intestinal peptide increased the levels of Gs alpha Go alpha, Gi-2 alpha, Gi-3 alpha, Gx alpha, G beta 36 and mRNAs. The effect of vasoactive intestinal peptide was however earlier and more pronounced. Gi-2 alpha mRNA levels showed the quantitatively largest alterations. Somatostatin upregulated Gs alpha and downregulated Go alpha and Gi-2 mRNAs. G protein mRNAs for Gi-2 alpha and Go alpha were increased by exposure of the cells to a medium devoid of serum. We conclude that G protein mRNA levels are subjected to alterations by hormones that act through the corresponding G proteins in the regulation of prolactin synthesis and secretion.     

Regulation of G protein mRNA levels by thyroliberin,vasoactive intestinal peptide and somatostatin in prolactin-producing rat pituitary adenoma cells

We have investigated the regulation of mRNA levels of alpha- and beta-subunits of guanine nucleotide-binding regulatory proteins (G proteins) by peptide hormones in prolactin producing rat pituitary adenoma cells (GH₃ cells) in culture. The cells were treated with thyroliberin (1 μm), vasoactive intestinal peptide (1 μm) or somatostatin (10 μM) for 6 to 48 hours. Thyroliberin and vasoactive intestinal peptide increased the levels of G_sα G_oα, G_i-2α, G_i-3α, G_xα, Gβ₃₆ and Gβ₃₅ mRNAs. The effect of vasoactive intestinal peptide was however earlier and more pronounced. G_i-2α mRNA levels showed the quantitatively largest alterations. Somatostatin upregulated G_sα and downregulated G_oα and G_i-2 mRNAs. G protein mRNAs for G_i-2α and G_oα were increased by exposure of the cells to a medium devoid of serum. We conclude that G protein mRNA levels are subjected to alterations by hormones that act through the corresponding G proteins in the regulation of prolactin synthesis and secretion.     

Unique pattern of cleavage of vasoactive intestinal peptide by human lymphocytes.

Human cultured T lymphocytes of the Jurkat line and myeloma cells of the U266 line cleaved the 28 amino acid vasoactive intestinal peptide (VIP1-28) preferentially at three sites with time- and temperature-dependence. The fragments VIP4-28 and VIP23-28) from an endopeptidase activity, and VIP15-28 from a trypsin-like peptidase, together represented a range of 26-65% of the VIP1-28 recovered after 2 hr at 37 degrees C or 4 hr at 22 degrees C, based on the absorbance of purified peptides and the radioactivity of [125I]Tyr10 VIP1-28. The endopeptidase activity was associated with membranes recovered after disruption of U266 cells by nitrogen cavitation. Pretreatment of intact U266 and Jurkat cells with diisopropylfluorophosphate (DFP) and the subsequently isolated subcellular particles with phenylmethylsulphonylfluoride (PMSF) and leupeptin inhibited the trypsin-like enzyme by a mean of 80%, without suppressing endopeptidase activity. In contrast, 0.1 mM DL-thiorphan and phosphoramidon blocked selectively a range of 35-70% of the endopeptidase activity in membrane preparations and intact cells. The capacity of lymphocytes to degrade VIP1-28 may substantially alter the effects of this neuromediator on functions of some subsets of T and B cells.     

Induction of aggregation of Raji human B-lymphoblastic cells by vasoactive intestinal peptide.

Subsets of neurons in the thymic cortex, Peyer's patches and lymphoid tissues of the respiratory system deliver vasoactive intestinal peptide (VIP) at nanomolar concentrations. The possible effects of VIP on B-cell adhesiveness in these tissues were examined in studies of the homotypic aggregation (HA) of human B-lymphoblastoid cells of the Raji line, which express a mean of 27,950 VIP receptors/cell with a mean Kd of 0.8 nM. Mean HA, assessed microscopically, attained a maximum of 54% after 8 hr with 0.1 microgram/ml of phorbol 12-myristate 13-acetate (PMA) (P < 0.01) and 31% after 24 hr with 10(-8) M VIP (P < 0.05), as contrasted with 13% and 20% at the respective times in medium alone, and both stimuli also increased the mean size of aggregates. The presence of the phosphodiesterase inhibitor Ro 20-1724 permitted 10(-9) M VIP, which had no effect alone, to raise the mean cyclic AMP content of Raji cells by more than 10-fold and concurrently to elevate mean HA from 55% in medium alone at 48 hr to 70% and from 55% at 72 hr to 68% (P < 0.05 for both). Monoclonal antibodies to lymphocyte function-associated (LFA-1) adhesive protein and to intercellular adherence molecule-1 (ICAM-1) suppressed significantly the HA of Raji cells induced by VIP and PMA. The effects of VIP on compartmental immunity in the lungs and intestines thus may be mediated in part by increases in lymphocyte adhesiveness, which could contribute to the regional accumulation of specifically immunocompetent cells.     

Expression of neurotrophins in hippocampal interneurons immunoreactive for the neuropeptides somatostatin, neuropeptide-Y, vasoactive intestinal polypeptide and cholecystokinin.

Using a double detection method, which combines in situ hybridization for the detection of neurotrophin messenger RNA with immunocytochemistry against the neuropeptides somatostatin, neuropeptide Y, vasoactive intestinal polypeptide and cholecystokinin, we have analysed the expression of the neurotrophins, nerve growth factor, brain-derived neurotrophic factor and neurotrophin-3, in distinct populations of neuropeptide-immunoreactive hippocampal interneurons. Nerve growth factor messenger RNA expression was found in subsets of the four subpopulations of neuropeptide-immunoreactive interneurons. The highest degree of co-localization was observed in the neuropeptide-Y-positive cells (up to 70%) and in somatostatin-immunoreactive cells (48%). Only small subsets of cholecystokinin- and vasoactive intestinal polypeptide-positive neurons (21% and 10%, respectively) displayed nerve growth factor hybridization signals. In contrast, expression of neurotrophin-3 messenger RNA was exclusively observed in 26% of neuropeptide-Y-immunoreactive cells. Brain-derived neurotrophic factor hybridization signals were never detected in the neuropeptide-positive hippocampal interneurons. Morphological analysis of neuropeptide-immunoreactive interneurons that express or lack nerve growth factor messenger RNA revealed that most perisomatic inhibitory neurons, such as large vasoactive intestinal polypeptide/ cholecystokinin-immunoreactive cells, showed positive nerve growth factor hybridization signals. In addition, some somatostatin/neuropeptide-Y-immunoreactive interneurons, which are responsible for dendritic inhibition of principal hippocampal neurons, expressed nerve growth factor messenger RNA. In contrast, interneurons specialized to innervate other GABAergic cells, such as small vasoactive intestinal polypeptide-positive cells, lacked nerve growth factor expression. All these data indicate that expression of neurotrophins is differentially regulated in functionally distinct classes of hippocampal interneurons immunoreactive for neuropeptides. We also analysed whether neuropeptide-immunoreactive interneurons expressing neurotrophins were targets of the GABAergic septohippocampal pathway. We used a triple detection method, combining anterograde tracing of this connection, with in situ hybridization for the detection of neurotrophin mRNA, and immunocytochemistry against neuropeptides. Our data showed that the four populations of hippocampal interneurons studied (somatostatin, neuropeptide-Y, vasoactive intestinal polypeptide and cholescystokinin) received GABAergic afferents from the septum. However, no preference for neuropeptide-immunoreactive cells expressing neurotrophins was observed, compared to neuropeptide-positive neurons lacking neurotrophin expression.     

Melatonin potentiates cyclic AMP production stimulated by vasoactive intestinal peptide in human lymphocytes.

The present paper demonstrates the effect of melatonin on cyclic AMP production in human lymphocytes from peripheral blood. Melatonin by itself did not influence cyclic AMP accumulation in these cells at any dose studied; however, the drug potentiated the effect of vasoactive intestinal peptide (VIP) on the cyclic nucleotide production. In the presence of physiological concentrations of VIP (either 1, 10 or 100 pM), melatonin potentiated cyclic AMP production. However, at high doses of VIP (either 1, 10 or 100 nM), melatonin exhibited no such effect. The results suggest that human lymphocytes are a target for melatonin and that it may participate, jointly with VIP, in the regulation of immune function.     

胎儿小肠生长抑素、胃泌素及血管活性肠肽免疫反应细胞的个体发生

目的：探讨生长抑素（SS）、胃泌素（Gas）、血管活性肠肽（VIP）免疫反应（IR）细胞在胎儿小肠中的个体发生及其分布。方法：免疫组织化学方法和图像分析技术。结果：SS-IR细胞和Gas-IR细胞于11周即可见于小肠三段绒毛上皮和尚未分化完全的肠腺细胞间。随着胎龄增长,SS-IR细胞数量和细胞内SS由少至多,其中以十二指肠细胞最多,回肠最少。Gas-IR细胞和细胞内的Gas在十二指肠中随胎龄增长逐渐增多;21周后空肠中则有所减少,回肠中则消失。VIP-IR细胞数量少,整个胎期细胞数量、形态及免疫染色强度均未见明显变化。结论：SS、Gas和VIP在胎儿小肠的内分泌细胞中表达,提示内分泌激素在胎儿小肠的发育过程中起调节作用。     

Myocardial vasoactive intestinal peptide and fibrosis induced by nitric oxide synthase inhibition in the rat

AIMS: In both normotensive and hypertensive rats, the degree of myocardial fibrosis is inversely correlated with the concentration of vasoactive intestinal peptide (VIP) in the myocardium. Treatment with nitric oxide (NO) synthase inhibitors also causes myocardial fibrosis. In this study, we sought to determine whether the myocardial fibrosis induced by treatment with the NO synthase inhibitor N(G)-nitro-l-arginine methyl ester (l-NAME) was also associated with depletion of VIP in the myocardium. METHODS: Male Wistar Kyoto (WKY) and spontaneous hypertensive rats (SHR) rats treated with l-NAME were randomized to low, intermediate or high salt content diets. After 4 weeks, the hearts were harvested, the degree of fibrosis quantified and VIP concentration measured. RESULTS: In WKY, systolic blood pressure increased with increasing dietary sodium (P < 0.05). Myocardial fibrosis also increased with increasing dietary sodium (P < 0.005). Myocardial VIP concentration decreased with increasing dietary sodium (P < 0.025). In contrast, in the SHR treated with l-NAME, systolic blood pressure increased but the increase was not affected by sodium intake. Further, myocardial fibrosis and myocardial VIP were unchanged by increased dietary sodium. Higher doses of l-NAME in the SHR did not increase the systolic blood pressure, increase the degree of myocardial fibrosis or decrease the myocardial concentration of VIP. These differences in myocardial VIP concentration may reflect differing effects of l-NAME on VIP metabolism, as l-NAME increased VIP metabolism in the WKY (P < 0.05) but did not change VIP metabolism in the SHR. CONCLUSIONS: We conclude that depletion of VIP in the myocardium is associated with increasing myocardial fibrosis in l-NAME treated WKY. As VIP depletion occurs in other models of myocardial fibrosis, it appears to be a common mechanism. Myocardial VIP depletion may therefore be a new and important factor in the pathogenesis of cardiac fibrosis.     

Release of somatostatin,neurotensin and vasoactive intestinal peptide upon inhibition of gastric acid secretion by duodenal acid and hyperosmolal solutions in the conscious rat

The inhibitory effect of duodenal exposure to acid and hyperosmolal solutions on pentagastrin-stimulated gastric acid secretion was studied in conscious rats equipped with chronic gastric fistula and duodenal Thiry-Vella loop. The loop was challenged with saline, HCl or hyperosmolal polyethylene glycol. Gastric acid secretion was measured in samples from the gastric fistula. Gut peptide concentrations were measured in duodenal perfusates collected each 30 min, and in plasma samples collected both during stimulated acid secretion alone, and at the end of experiments in combination with luminal challenges of the loops. During pentagastrin-stimulated gastric acid secretion, luminal perfusion of the duodenal loop with acid caused inhibition of acid secretion (P < 0.001) and a prominent release of somatostatin both to the lumen (P < 0.001) and to the circulation (P < 0.05). Also, neurotensin (P < 0.01) and vasoactive intestinal peptide (P < 0.01) were released to the lumen, but not to the circulation. Upon perfusion of the duodenal loop with hyperosmolal polyethylene glycol, acid secretion was inhibited (P < 0.05) and somatostatin alone was released to the luminal side (P < 0.01). In conclusion, duodenal exposure to acid inhibits pentagastrin-stimulated gastric acid secretion and releases SOM to the circulation that may directly inhibit acid secretion. Concomitantly, somatostatin (SOM), neurotensin and vasoactive intestinal peptide are released to the lumen. Duodenal exposure to hyperosmolal polyethylene glycol inhibits acid secretion with a luminal release of SOM only. Thus, luminal acid and hyperosmolal solutions inhibit gastric acid secretion by separate mechanisms. After acid or hyperosmolal challenge, the release of SOM to the circulation indicates gastric acid inhibition in an endocrine manner, while a luminal release of gut peptides indicates a local peptide overflow that might be of importance via paracrine regulatory mechanisms in the intact animal.     

Stimulation of insulin and glucagon secretion by vasoactive intestinal peptide.

In vivo, vasoactive intestinal peptide (VIP) produces simultaneous increases in blood glucose and insulin levels. In order to determine whether VIP, like its homologues, also stimulates insulin secretion directly, studies were made in controlled glucose media employing the vascularly perfused cat pancreas. VIP stimulated insulin secretion significantly in the presence of constant physiological concentrations of glucose. The highest insulin response to VIP (100.3+/-8.1 muU/min) approached the highest insulin response to glucose (119.9 +/- 12.0 muU/min). In the absence of glucose, the insulin response to VIP was insignificant. Unexpectedly, VIP was found to be a more effective stimulant of glucagon than of insulin secretion. The highest glucagon response to VIP (327+/-51% of control levels) was attained in the presence of physiological concentrations of glucose and equalled the glucagon response obtained upon withdrawal of glucose from the perfusate. The glucagon response to VIP was blocked by increasing the glucose in the perfusate. These studies indicate the VIP present in pancreatic islets might play a role in the local control of pancreatic endocrine function.     

Pharmacologically distinct vasoactive intestinal peptide binding sites: CNS localization and role in embryonic growth.

In vitro autoradiography with [125I]vasoactive intestinal peptide revealed that the vasoactive intestinal peptide analogue, stearyl-norleucine17 vasoactive intestinal peptide, reported to be inactive at adenylyl cyclase-linked receptors in astrocytes, displaced a subset of vasoactive intestinal peptide binding sites on rat brain sections. These sites were widespread in adult rat brains and enriched in the olfactory bulb and thalamus, and corresponded to previously demonstrated GTP-insensitive vasoactive intestinal peptide binding sites. Stearyl-norleucine17 vasoactive intestinal peptide also identified receptors in rat lung and liver. In the adult brain, the stearyl-norleucine analog displaced only GTP-insensitive vasoactive intestinal peptide binding sites. In contrast, stearyl-norleucine17 vasoactive intestinal peptide-displaceable sites in the embryonic day 9 mouse appeared to include both GTP-sensitive and GTP-insensitive binding sites. This observation suggested the presence of an embryonic vasoactive intestinal peptide receptor with distinct pharmacological properties. Treatment of whole cultured mouse embryos with stearyl-norleucine17 vasoactive intestinal peptide resulted in stimulation of embryonic growth, with the stearyl-norleucine analog equipotent to vasoactive intestinal peptide, but less efficacious at higher concentrations (10(-7) M). Embryonic growth was inhibited by pituitary adenylyl cyclase-activating peptide and 8-bromoadenosine 3',5'-cyclic monophosphate. In addition, 8-bromoadenosine 3',5'-cyclic monophosphate inhibited stearyl-norleucine17 vasoactive intestinal peptide-stimulated growth. The results of the current study support the hypothesis that vasoactive intestinal peptide regulation of early postimplantation embryonic growth occurs, at least in part, independently of adenylyl cyclase stimulation.     

Influence of beta-endorphin, somatostatin, substance P and vasoactive intestinal peptide on the proliferative response of human peripheral blood T lymphocytes to mercuric chloride

The influence of beta-endorphin, somatostatin, substance P (SP) and vasoactive intestinal peptide (VIP) was tested on the proliferative response to mercuric chloride of human peripheral blood T lymphocytes, cultured for 5 days. When beta-endorphin, 10(-8) M, was added 1 h after mercuric chloride, there was an enhancement of the response, while a slight suppression was obtained with a 10(-6) M concentration of SP and VIP. When beta-endorphin, 10(-7)-10(-9) M, somatostatin, 10(-6)-10(-9) M, and SP, 10(-11)-10(-12) M, were added 3 days after mercuric chloride, they enhanced the response. At 10(-6) M, SP gave a suppressive effect.     

Immunohistochemical localization of receptors for vasoactive intestinal peptide and substance P in human trachea.

BACKGROUND: Tissue sections of human cervical trachea were processed for immunohistochemical demonstration of receptors for substance P [using an anti-SP anti-idiotypic antiserum directed toward the ligand binding site of the receptor (Couraud J-Y, Escher ED, Regoli D, Imhof V, Rossignol B, Pradelles P. Anti-substance P anti-idiotypic antibodies: Characterization and biological activities. J Biol Chem 1985;260:9461-9; Couraud J-Y, Maillet S, Grassi J, Frobert Y, Pradelles P. Characterization and properties of anti-substance P antiidiotypic antibodies. Methods Enzymol 1989; 178:275-300)] and vasoactive intestinal peptide (VIP; utilizing a monoclonal antibody toward VIP receptors of an adenocarcinoma cell line (Pichon J, Hirn M, Muller J-M, Mangeat P, Marvaldi J. Anticell surface monoclonal antibodies which antagonize the action of VIP in a human adenocarcinoma cell line (HT29). EMBO J 1983;2:1017-22)], respectively. Mucus cells of the submucosal glands (identified by periodic acid Schiff staining) and neuroendocrine cells of the respiratory epithelium (identified by immunoreactivity to protein gene product 9.5) displayed intense VIP receptor-immunoreactivity. Other tissue components known to respond to exogenously administered VIP, e.g., trachealis muscle, lacked VIP receptor-immunoreactivity, indicating that the monoclonal antibody did not label all receptor subtypes. In accordance with the known pharmacological actions of substance P upon the airways, the anti-substance P receptor antibody labeled the trachealis muscle, submucosal glands, and respiratory epithelium, predominantly at the luminal aspect. Since substance P as well as the structurally related tachykinin, neurokinin A, competed with the anti-receptor antibody in binding to the tissue section, it is likely that both NK-1 and NK-2 receptor subtypes were labeled. The present histochemical approach to localize peptide receptors in the trachea allowed precise analysis of distribution unreached by previous studies using autoradiography. Together with pharmacological data, these morphological findings contribute to the understanding of the sequences of events evoked by the neuropeptides, substance P and VIP, in the human trachea.     

Co-expression patterns of the neuropeptides vasoactive intestinal peptide and cholecystokinin with the transduction molecules alpha-gustducin and T1R2 in rat taste receptor cells

Taste receptor cells are primary sensory receptors utilized by the nervous system to detect the presence of gustatory stimuli in the oral cavity. These cells are particularly heterogeneous and may be divided into various subtypes based on morphological, histochemical, or physiological criteria. One example is the heterogeneous expression of neuropeptides, such as cholecystokinin and vasoactive intestinal polypeptide. These peptides are hypothesized to participate in the transduction processes. To pursue examination of this hypothesis, this study explored the relationship of peptide expression with two important and mostly non-overlapping transductive elements--the taste-specific G protein gustducin, involved in bitter and sweet transduction cascades, and the seven transmembrane taste receptor T1R2, hypothesized to respond to sweet compounds. Double labeling experiments were performed on taste buds of the posterior rat tongue combining immunocytochemistry for peptide expression and in situ hybridization experiments for either gustducin or T1R2 expression. Additionally, vasoactive intestinal peptide (VIP)-expression in posterior taste receptor cells was confirmed using the technique of RT-PCR. More than half (56%) of the CCK-expressing taste receptor cells co-expressed alpha-gustducin mRNA whereas far fewer (15%) co-expressed T1R2 mRNA. A majority of VIP-expressing taste receptor cells co-expressed alpha-gustducin mRNA (60%) whereas only 19% of these cells co-expressed T1R2 mRNA. More remarkable was the observation that these two peptides displayed almost identical expression patterns with these signal transduction molecules, suggesting that peptides are not randomly expressed with relation to signal transduction molecules. This observation supports the hypothesis that peptides may play roles in transduction. Further physiological exploration will be required to elucidate the nature of these roles.     

Regulation of inhibitory synaptic transmission by vasoactive intestinal peptide (VIP) in the mouse suprachiasmatic nucleus

Circadian rhythmicity in mammals is generated by a pair of nuclei in the anterior hypothalamus known as the suprachiasmatic nuclei (SCN), whose neurons express a variety of neuropeptides that are thought to play an important role in the circadian timing system. To evaluate the influence of VIP on inhibitory synaptic transmission between SCN neurons, we used whole cell patch-clamp recording in an acute brain slice preparation of mouse SCN. Baseline spontaneous GABAergic inhibitory postsynaptic currents (IPSCs) varied significantly between regions and across phases, with a greater frequency of IPSCs observed in the dorsomedial region during the early night. Bath-applied VIP caused a significant increase in the frequency of spontaneous inhibitory postsynaptic currents (sIPSC) in a reversible and dose-dependent manner with no effect on the mean amplitude or kinetic parameters. The effect of VIP was widespread throughout the SCN and observed in both ventrolateral (VL) and dorsomedial (DM) regions. In the presence of tetrodotoxin, VIP increased the frequency of miniature IPSCs without affecting the mean magnitude or kinetic parameters. The magnitude of the enhancement by VIP was significantly larger during the day than during the night. Pretreatment with the VIP-PACAP receptor antagonist [Ac-Tyr1, D-Phe2]-GHRF 1-29 or the selective VPAC2 receptor antagonist PG 99-465 completely blocked the VIP-induced enhancement. The effect of VIP appears to be mediated by a cAMP/PKA-dependent mechanism as forskolin mimics, while the PKA antagonist H-89 blocks the observed enhancement of GABA currents. Our data suggest that VIP activates presynaptic VPAC2 receptors to regulate inhibitory synaptic transmission within the SCN and that this effect varies from day to night.     

Localization of vasoactive intestinal peptide immunoreactivity in human foetus and newborn infant spinal cord

Using an indirect immunofluorescence method the distribution of vasoactive intestinal peptide (VIP) immunoreactivity was studied in human foetus and newborn infant spinal cord and dorsal root ganglia. Further, for comparison some newborn infant brains were also investigated. Vasoactive intestinal peptide-like immunoreactive fibres were exclusively found in the caudal spinal cord and corresponding dorsal root ganglia. No immunoreactive cell bodies were detected. The first appearance of VIP-like immunoreactive fibres in both spinal cord and dorsal root ganglia was suggested during the fourth month of foetal life. Most immunolabelled fibres, concentrated in the sacral segment, were distributed in the Lissauer tract, along the dorsolateral gray border, in the intermediolateral areas and near the central canal in the dorsolateral commissure. A few VIP-like immunoreactive fibres were also seen in the dorsal funiculus and occasionally in the ventral gray horn and ventral roots. Further, a large population of VIP-like immunoreactive fibres occurs longitudinally in dorsal root, in ganglia and in the spinal nerve exit zone. These findings indicate the early appearance of VIP-like immunoreactive fibres in the human foetus spinal cord and corresponding ganglia. Moreover, they emphasize that in both foetus and newborn infant spinal cord VIP-like immunoreactive fibre distribution is limited to the lumbosacral segment.     

Comparison of vasoactive intestinal peptide and secretin in stimulation of pancreatic secretion.

Pancreatic volume flow as well as bicarbonate and protein secretion have been measured in chronic pancreatic fistula cats and dogs in response to I.V. infusion of VIP and secretin or duodenal perfusion of sodium oleate and HCl solution. 2. VIP and secretin infused I.V. in cats produced superimposable pancreatic dose-response curves for volume flow and bicarbonate secretion, reaching almost identical observed and maximal calculated outputs with both peptides. In dogs, VIP was shown previously to be a much less effective stimulant of pancreatic secretion than secretin and the maximal observed bicarbonate output in response to VIP was only about 17% of that to secretin (Konturek, Thor, Dembinski & Król, 1975). It is condluded that VIP in cats is a secretin-like full agonist, whereas in dogs it is a partial agonist of pancreatic bicarbonate secretion. 3. In cats, secretin and VIP showed equal efficacy and their combination exhibited an augmentatory action on pancreatic bicarbonate secretion with additive kinetics, whereas in dogs, VIP was found to have a lower efficacy than secretin and to inhibit competitively secretin-induced pancreatic secretion. These results might be explained by the interaction of VIP and secretin, two chemically related peptides, on a common receptor site of the exocrine pancreas. 4. Caerulein, an analogue of CCK-PZ, infused I.V. in cats and dogs caused a negligible pancreatic bicarbonate secretion and a potent dose-dependent protein secretion. The combination of graded doses of VIP or secretin with a background dose of caerulein resulted in significantly higher bicarbonate and protein outputs than those induced by VIP or secretin alone. 5. Duodenal perfusion of sodium oleate soap in cats and dogs produced pancreatic dose-response curves for volume flow and bicarbonate output similar to those evoked by VIP in these species. Pancreatic protein secretion in response to luminal oleate was slightly higher than could be accounted for by the action of VIP alone. This might be attributed to the release by oleate not only of endogenous VIP but also CCK-PZ or to the vago-vagal reflexes from gut to pancreas. The results of our combined study on cats and dogs suggest the possibility that oleate releases VIP from the gut and that this peptide may play a physiological role in the stimulation of pancreatic secretion.