Release Of Gastrin-releasing Peptide Research Articles

Immediate Release of Gastrin-Releasing Peptide Mediates Delayed Radiation-Induced Pulmonary Fibrosis

Radiation-induced pulmonary fibrosis (RTPF) is a progressive, serious condition in many subjects treated for thoracic malignancies or after accidental nuclear exposure. No biomarker exists for identifying the irradiated subjects most susceptible to pulmonary fibrosis (PF). Previously, we determined that gastrin-releasing peptide (GRP) was elevated within days after birth in newborns exposed to hyperoxia who later developed chronic lung disease. The goal of the current study was to test whether radiation (RT) exposure triggers GRP release in mice and whether this contributes to RTPF invivo. We determined urine GRP levels and lung GRP immunostaining in mice 0 to 24 after post-thoracic RT (15 Gy). Urine GRP levels were significantly elevated between 24 hours post-RT; GRP-blocking monoclonal antibody 2A11, given minutes post-RT, abrogated urine GRP levels by 6 to 12 hours and also altered phosphoprotein signaling pathways at 24 hours post-RT. Strong extracellular GRP immunostaining was observed in lung at 6 hours post-RT. Mice given one dose of GRP monoclonal antibody 2A11 24 hours post-RT had significantly reduced myofibroblast accumulation and collagen deposition 15 weeks later, indicating protection against lung fibrosis. Therefore, elevation of urine GRP could be predictive of RTPF development. In addition, transient GRP blockade could mitigate PF in normal lung after therapeutic or accidental RT exposure.

Activation of gastrin-releasing peptide receptors at the infralimbic cortex elicits gastrin-releasing peptide release at the basolateral amygdala: Implications for conditioned fear

The basolateral amygdala (BLA) and infralimbic (IL) cortex share strong reciprocal interconnections and are key structures in conditioned fear circuitry. Gastrin-releasing peptide (GRP) or its receptor antagonists can modulate the conditioned fear response when exogenously administered at either of these sites, and increased release of GRP at the BLA occurs in response to conditioned fear recall. The present study sought to determine whether a functional pathway utilizing GRP exists between the IL cortex and BLA and whether this pathway is also influenced by amygdala corticotropin-releasing factor (CRF) release. To this end, we assessed the effects of intra-IL cortex injection of GRP or GRP co-administered with a receptor antagonist, RC-3095, on the downstream release of GRP and/or CRF at the BLA. Results showed that microinjection of GRP at the IL cortex increased the release of GRP, but not CRF, at the BLA, an effect blocked by co-administration of RC-3095. Administration of RC-3095 into the IL cortex on its own, however, also elicited the release of GRP (but not CRF) at the BLA. These findings suggest that a functional pathway utilizing GRP (among other factors) exists between the IL cortex and BLA that may be relevant to conditioned fear, but that GRP and CRF do not interact within this circuitry. Moreover, the finding that the release profile of GRP was similar following administration of either GRP or its receptor antagonist, lends support to the view that RC-3095 has partial agonist properties. Together these findings provide further evidence for the involvement of GRP in fear and anxiety-related disorders.

Roles of light and serotonin in the regulation of gastrin‐releasing peptide and arginine vasopressin output in the hamster SCN circadian clock

Daily timing of the mammalian circadian clock of the suprachiasmatic nucleus (SCN) is regulated by photic input from the retina via the retinohypothalamic tract. This signaling is mediated by glutamate, which activates SCN retinorecipient units communicating to pacemaker cells in part through the release of gastrin-releasing peptide (GRP). Efferent signaling from the SCN involves another SCN-containing peptide, arginine vasopressin (AVP). Little is known regarding the mechanisms regulating these peptides, as literature on in vivo peptide release in the SCN is sparse. Here, microdialysis-radioimmunoassay procedures were used to characterize mechanisms controlling GRP and AVP release in the hamster SCN. In animals housed under a 14/10-h light-dark cycle both peptides exhibited daily fluctuations of release, with levels increasing during the morning to peak around midday. Under constant darkness, this pattern persisted for AVP, but rhythmicity was altered for GRP, characterized by a broad plateau throughout the subjective night and early subjective day. Neuronal release of the peptides was confirmed by their suppression with reverse-microdialysis perfusion of calcium blockers and stimulation with depolarizing agents. Reverse-microdialysis perfusion with the 5-HT(1A,7) agonist 8-OH-DPAT ((±)-8-hydroxydipropylaminotetralin hydrobromide) during the day significantly suppressed GRP but had little effect on AVP. Also, perfusion with the glutamate agonist NMDA, or exposure to light at night, increased GRP but did not affect AVP. These analyses reveal distinct daily rhythms of SCN peptidergic activity, with GRP but not AVP release attenuated by serotonergic activation that inhibits photic phase-resetting, and activated by glutamatergic and photic stimulation that mediate this phase-resetting.

Effects of corticosterone on corticotrophin‐releasing hormone and gastrin‐releasing peptide release in response to an aversive stimulus in two regions of the forebrain (central nucleus of the amygdala and prefrontal cortex)

Previous research has shown that chronic corticosterone treatment increases the expression of corticotrophin-releasing hormone (CRH) mRNA at the central nucleus of the amygdala (CeA). Like CRH, gastrin-releasing peptide (GRP) appears to be involved in mediation of the stress response and is released at the CeA during exposure to an acute stressor. Using in-vivo microdialysis, this study examined the effects of corticosterone treatment on the release of CRH and GRP in response to an airpuff challenge at two forebrain regions, the CeA and medial prefrontal cortex. Adrenally intact rats were treated with corticosterone by systemic implants over a 14-day period prior to microdialysis probe insertion. We found that, at both regions, the airpuff-induced CRH and GRP release were enhanced in the corticosterone pellet-implanted rats as compared with the release observed in the vehicle-implanted control rats. These findings suggest that chronic corticosterone exposure potentiates the stressor-elicited release of CRH and GRP. As cortisol dysregulation has frequently been reported in people with psychiatric conditions, such as anxiety disorders or depression, a better understanding of the glucocorticoid-mediating regulation of CRH and GRP may provide insight into the underlying neurochemical mechanisms involved in both adaptive fear-type responses and maladaptive responses leading to pathology.

Gastrin-releasing peptide is a modulatory neurotransmitter of the descending phase of the peristaltic reflex.

The physiological role of gastrin-releasing peptide (GRP) and of its cognate receptors in regulating the intestinal peristaltic reflex was examined in a three-compartment flat-sheet preparation of rat colon. Mucosal stimulation applied to the central compartment at high, but not low levels of intensity, induced GRP release in the caudad compartment where descending relaxation was measured, but not into the ascending compartment where ascending contraction was measured or into the central compartment where the stimuli were applied. The selective GRP (BB(2)) receptor antagonist, [D-Phe(6),des-Met(14)]bombesin(6-14), inhibited descending relaxation and VIP release in the caudad compartment induced by high but not by low levels of stimulation applied to the mucosa in the central compartment. The selective neuromedin B (BB(1)) receptor antagonist, BIM-23127, had no effect on descending relaxation or VIP release. Neither the BB(1) nor the BB(2) antagonist had any effect on ascending contraction or substance P release in the orad compartment. Consistent with the effects of the antagonists on the peristaltic reflex, the BB(2) antagonist but not the BB(1) antagonist decreased the velocity of propulsion of artificial fecal pellets through isolated segments of guinea pig distal colon. The results indicate that GRP is selectively released from myenteric neurons in descending pathways during the peristaltic reflex and that it acts via BB(2) receptors to augment the descending phase of the peristaltic reflex and propulsion.

Ultrastructural localization of gastrin-releasing peptide (GRP) in the uterine gland of cow.

Gastrin-releasing peptide (GRP) is thought to act mainly as a neurotransmitter and localized almost exclusively to neurons and neuroendocrine cells. Recently, the localization of GRP in mammalian uterus and placenta has been demonstrated. Moreover, the exocrine manner of GRP release was deduced in ewes from the distribution of GRP on the uterine gland cells and its secretion as well as in the circulation. However, these reports have been examined at light-microscopic level. The present study was designed to make clear the localization of GRP in the uterine gland cells of nonpregnant and pregnant cows using an avidin-biotin-peroxidase complex (ABC) method at light-microscopic level and a pre-embedding immunogold with silver enhancement method at electron-microscopic level. The light-microscopic observation showed positive staining for GRP immunoreactivity in the supranuclear region and in the secreted materials of the uterine gland cells. At the electron-microscopic level, the supranuclear secretory granules and the secreted materials on the surface of the cell were labeled with immunogold particles representing GRP immunoreactivity in the uterine gland cells of nonpregnant and pregnant cows. Western blotting analysis showed a larger molecular form of GRP in the endometrial tissues taken from nonpregnant and pregnant cows. The present results revealed the localization of GRP in the uterine gland cells at light- and electron-microscopic levels and suggested the release of GRP from the cell into the lumen of the gland by exocrine manner.

Role of GRPergic neurons in secretin-evoked exocrine secretion in isolated rat pancreas.

Effects of intrapancreatic gastrin-releasing peptide (GRP)-containing neurons on secretin-induced pancreatic secretion were investigated in the totally isolated perfused rat pancreas. Electrical field stimulation (EFS) increased secretin (12 pM)-induced pancreatic secretions of fluid and amylase. EFS induced a twofold increase in GRP concentration in portal effluent, which was completely inhibited by tetrodotoxin but not modified by atropine. An anti-GRP antiserum inhibited the EFS-enhanced secretin-induced secretions of fluid and amylase by 12 and 43%, respectively, whereas a simultaneous infusion of the antiserum and atropine completely abolished them. Exogenous GRP dose-dependently increased the secretin-induced pancreatic secretion with an additive effect on fluid secretion and a potentiating effect on amylase secretion, which was not affected by atropine. In conclusion, excitation by EFS of GRPergic neurons in the isolated rat pancreas results in the release of GRP, which exerts an additive effect on fluid secretion and a potentiating effect on amylase secretion stimulated by secretin. The release and action of GRP in the rat pancreas are independent of cholinergic tone.

Evidence that endogenous GRP in rat stomach mediates omeprazole-induced hypergastrinemia.

To investigate the physiological role of endogenous gastrin-releasing peptide (GRP) in regulating the release of gastrin, we evaluated the response of intragastric pH, gastrin, and GRP after omeprazole treatment in rats. A significant elevation of the plasma level of GRP (P < 0.01) and a significant reduction of the antral content of GRP (P < 0.05) were observed after the administration of 100 mg/kg omeprazole. The antral content of GRP was significantly decreased 12 h after omeprazole administration and thereafter gradually returned to control levels. Peak values for intragastric pH and plasma GRP were observed 3 h after omeprazole administration and before the peak of serum gastrin. The bombesin antagonist [D-Phe6]-bombesin-(6,13)-methyl ester significantly inhibited gastrin release after omeprazole treatment (P < 0.05). These observations indicate that omeprazole-induced inhibition of acid secretion stimulates the release of GRP and suggest that the secretion of GRP induced by omeprazole may stimulate the secretion of gastrin, at least in the early phase.

Pretransplant clinicopathological correlation in end-stage primary pulmonary hypertension

The aim of the study was to see if there was any correlation between the histopathology, ultrastructure, pulmonary endocrinology and clinical manifestations of end-stage primary pulmonary hypertension. Twenty patients undergoing heart-lung transplantation for the disease were studied. The nature and duration of symptoms and signs, results of haematological, electrocardiographic, radiographic, echocardiographic and haemodynamic studies, and the response of patients to vasodilators were compared with data from histopathological and ultrastructural study of lungs removed at transplantation. Length of clinical history and clinical evidence of severe disease were not necessarily associated with advanced histopathology, nor did the presence of small, contracted muscular pulmonary arteries imply responsiveness to vasodilators. Numbers of gastrin-releasing peptide-containing pulmonary endocrine cells were greater in lungs in which there was activity of myofibroblasts in pulmonary arterial vessels, and correlated negatively with mean pulmonary artery pressure and pulmonary artery systolic pressure. Whereas the prognosis of primary pulmonary hypertension cannot as yet be defined by other than its clinical manifestations, intimal proliferation as well as vasoconstriction may be important in its pathogenesis. The release of gastrin-releasing peptide from pulmonary endocrine cells may possibly be involved in this process.

&lt;b&gt;GASTRIN-RELEASING PEPTIDE (GRP)-CONTAINING NEURONS IN THE RAT OXYNTIC MUCOSA VVITH SPECIAL REFERENCE TO EXPERIMENTALLY INDUCED PEPTIC &lt;/b&gt;&lt;b&gt;ULCERS &lt;/b&gt;

The relation between gastrin-releasing peptide (GRP)-immunoreactive nerves and development of stomach ulcers was studied in rats. The ulcer was induced by exposure of animals to restraint plus water immersion or by administration of insulin, serotonin, aspirin, indomethacin or ethanol. Immunohistochemistry demonstrated a network of nerves exhibiting the immunoreactivity for GRP in the oxyntic mucosa of normal animals. A marked decrease in number of the GRP-immunoreactive nerves was seen to accompany with the ulcers induced by the restraint stress or by insulin. On the other hand, no appreciable change in number of the GRP-immunoreactive nerves was found in ulcers induced by indomethacin, serotonin, ethanol or aspirin. Vagotomy prevented not only the development of ulcers by stress or insulin, but also the decrease in the GRP-immunoreactive nerves. Electrical stimulation of the vagal nerve resulted in ulcer formation and also caused a marked decrease in number of the GRP-immunoreactive nerves in the oxyntic mucosa. The results suggest the possibility that depletion of GRP from nerves in the oxyntic mucosa may be involved in the development of stress or insulin ulcers and that such a release of GRP may be caused by hyperactivity of the vagal nerve.

GRP nerves in pig antrum: role of GRP in vagal control of gastrin secretion

We extracted gastrin-releasing peptide (GRP) and its C-terminal decapeptide corresponding to 6.4 and 6.8 pmol/g from pig antrum mucosa. By immunohistochemistry GRP was localized to mucosal, submucosal, and myenteric nerve fibers. A few nerve cell bodies were also identified. Using isolated perfused pig antrum with intact vagal innervation, we found concomitant, atropine-resistant release of GRP and gastrin during electrical stimulation of the vagal nerves. Intra-arterial GRP at 10(-11)-10(-10) mol/l caused up to fivefold, dose-dependent increases in gastrin secretion; higher doses were less effective and completely desensitized the gastrin cells for the lower doses. After desensitization, vagal stimulation no longer produced gastrin secretion. The substance P antagonist [D-Arg, D-Pro, D-Trp, Leu]-substance P, described as also antagonizing the actions of bombesin, decreased the gastrin response to GRP and abolished the effect of vagal stimulation. The available evidence strongly suggests that GRP nerves are responsible for the stimulatory vagal effects on gastrin secretion in the pig.

Vagal regulation of GRP, gastric somatostatin, and gastrin secretion in vitro.

The effect of electrical stimulation of the vagus nerves on the release of immunoreactive gastrin-releasing peptide (GRP), gastrin, and somatostatin was investigated using the isolated perfused rat stomach. Electrical stimulation (10 Hz, 1 ms duration, 10 V) of the peripheral end of the subdiaphragmatic vagal trunks produced a significant increase in both GRP and gastrin but a decrease in somatostatin. The infusion of atropine sulfate at a concentration of 10(-5) M augmented GRP release and reversed the decrease in somatostatin release in response to vagal stimulation to an increase above basal levels. However, the gastrin response to vagal stimulation was not affected by atropine. The infusion of hexamethonium bromide at a concentration of 10(-4) M significantly suppressed GRP release but did not affect gastrin secretion in response to vagal stimulation. On the other hand, the somatostatin response to vagal stimulation was completely abolished by hexamethonium. These findings lead us to conclude that the intramural GRP neurons might play an important role in the regulation of gastrin as well as somatostatin secretion and that somatostatin secretion may be controlled not only by a cholinergic inhibitory neuron but also by a noncholinergic, e.g., peptidergic stimulatory neuron, both of which may be regulated through preganglionic vagal fibers via nicotinic receptors. In addition, because the infusion of 10(-7) M GRP suppressed the somatostatin secretion, we suggest that either GRP should be excluded from the list of candidates for the noncholinergic stimulatory neurotransmitter for somatostatin secretion or that there are different mechanisms of action for endogenous and exogenous GRP.