Jiayuan Ren

Calcitonin Gene-Related Peptide Modulates Acid-Mediated Regulation of Somatostatin and Gastrin Release From Rat Antrum

BACKGROUND & AIMS Acid has been shown to stimulate calcitonin gene-related peptide (CGRP) release from peripheral sensory afferent nerve endings in the stomach. The aim of this study was to determine whether endogenous CGRP was involved, by a neurocrine mechanism, in acid-mediated stimulation of somatostatin and inhibition of gastrin release. METHODS A two-compartment sleeve of antral mucosal/submucosal tissue was perfused to determine sensory nerve and endocrine cell responses to luminal acid. CGRP receptor antagonist, CGRP8-37, was used to inhibit the actions of endogenously released CGRP. RESULTS Perfusion of the antral sleeve lumen with media of increasing hydrogen ion concentration caused pH-dependent increases in CGRP and somatostatin release and decrease in gastrin release. CGRP8-37 inhibited significantly basal somatostatin (-36%) and stimulated basal gastrin (+65%) release (P < 0.02). Furthermore, CGRP8-37 administration prevented luminal acid-mediated inhibition of gastrin release and stimulation of somatostatin release. These results indicate that CGRP8-37 prevented acid-mediated feedback inhibition of gastrin release and acid-induced feedforward somatostatin release. CONCLUSION These results suggest that CGRP plays an important role in the response of antral D and G cells to luminal acid and that local effector action of endogenous CGRP participates in regulation of antral regulatory peptide secretion.

Calcitonin gene-related peptide mediates capsaicin-induced neuroendocrine responses in rat antrum

BACKGROUND Calcitonin gene-related peptide (CGRP) is a 37-amino acid peptide localized to primary sensory afferent nerves in the rat stomach. The actions of CGRP in regulating antral neuroendocrine function were examined in vitro through the use of capsaicin, an agent capable of evoking neuropeptide release from peripheral sensory nerve endings. These results were compared with the effects of exogenous CGRP and CGRP antagonist, CGRP8-37. METHODS Rat antral mucosal/submucosal fragments were incubated in either static or dynamic perifusion experiments. Media were assayed for gastrin, somatostatin, CGRP, and acetylcholine. RESULTS Capsaicin, like exogenous CGRP, stimulated antral somatostatin release and inhibited both gastrin release and acetylcholine discharge. Low dose capsaicin (1 x 10(-5) mol/L) caused significant stimulation of CGRP release: 33 +/- 0.2 vs. 14 +/- 1 pg/mL protein; P < 0.001. Tetrodotoxin blocked capsaicin-induced inhibition of acetylcholine release and prevented partially capsaicin-mediated stimulation of CGRP release. The CGRP receptor antagonist CGRP8-37 prevented capsaicin-induced D-cell stimulation and inhibition of G-cell secretion and cholinergic discharge. CONCLUSIONS The effects of capsaicin-induced changes in antral D- and G-cell secretion and acetylcholine discharge are due primarily to release of CGRP. Antral CGRP release from primary sensory afferent nerve terminals may act as a local effector substance to regulate antral neuroendocrine function.

Presynaptic muscarinic receptors modulate acetylcholine release from rat antral mucosal/submucosal nerves.

The purpose of the present studies was to determine whether autoinhibition of acetylcholine release could be demonstratedin vitro from mucosal/submucosal neurons in rat antrum. Rat antral mucosal/submucosal tissues preloaded with [3H]choline were perifused and [3H]acetylcholine release measured under basal and stimulated conditions. Carbachol inhibited both spontaneous and evoked (electrical field stimulation, KCl) acetylcholine release from rat antral tissues: 1×10−5M carbachol inhibited basal [3H]ACh release maximally to −38.2±3.1% (P<0.001 vs control). The nonselective muscarinic antagonist atropine enhanced both basal and stimulated acetylcholine release and abolished carbachol-induced inhibition of acetylcholine release. Pirenzepine, a muscarinic M1 receptor antagonist, inhibited acetylcholine release and did not alter carbachol-induced inhibition of acetylcholine release. In conclusion, acetylcholine release from rat antral mucosal/submucosal neurons is regulated negatively by a presynaptic feedback mechanism involving M2 and/or M3 receptors, while presynaptic M1 receptors facilitate release of neurotransmitter.

Involvement of capsaicin-sensitive sensory neurons in stress-induced gastroduodenal mucosal injury in rats.

The pathogenesis of stress-induced gastroduodenal mucosal injury is complex and incompletely understood. The aim of this investigation was to examine the involvement of gastric and duodenal capsaicin-sensitive neurons in mucosal damage associated with water-restraint stress (WRS) in rats. Following WRS, gastroduodenal mucosal injury was quantitated by macroscopic and microscopic methods. Calcitonin gene-related peptide (CGRP) content was measured by radioimmunoassay. WRS-induced mucosal erosive injury in the stomach and duodenum (40.9 ± 4.2 and 5.1 ± 0.6 mm2, respectively) was reduced significantly (by 88% and 67%, respectively) by acute intragastric capsaicin administration prior to WRS. In contrast, sensory denervation by chronic capsaicin significantly increased the area of gastric injury and duodenal damage. WRS alone caused a significant reduction (by 52% and −35%, respectively) in gastric and duodenal CGRP content, which was prevented by acute capsaicin treatment. The data suggest that gastric and duodenal sensory neurons and CGRP are involved in the pathogenesis of stress-induced mucosal injury to the stomach and duodenum.

Effects of calcitonin gene-related peptide on somatostatin and gastrin gene expression in rat antrum.

The ability of exogenous calcitonin gene-related peptide (CGRP) to regulate gastric somatostatin and gastrin messenger RNA was studied in vitro in rat antral mucosal/submucosal tissues. Somatostatin and gastrin mRNA were quantified by Northern and dot blot hybridization and regulatory peptides were measured by radioimmunoassay. Incubation of antral tissues in the presence of CGRP [1 x 10(-7) M] for 60 min resulted in a reciprocal increase in somatostatin and a decrease in gastrin release: 214.7+/-28.5 vs. control of 81.7+/-5.9 pg somatostatin per gram of tissue and 2.2+/-0.3 vs. control of 5.5+/-0.7 ng gastrin per gram of tissue (P < 0.001). CGRP caused parallel changes in somatostatin and gastrin mRNA levels: somatostatin mRNA increased by 212% from 0.40+/-0.02 to 1.25+/-0.09 absorbance units (AU) (P < 0.001) and gastrin mRNA decreased by 73% from 0.55+/-0.08 to 0.15+/-0.02 AU (P < 0.001). Somatostatin monoclonal antibody prevented CGRP-mediated inhibition of both gastrin release and gastrin mRNA levels. In conclusion, CGRP is capable of modulating both the secretion and gene expression of regulatory peptides from antral G and D cells. Somatostatin immunoneutralization studies suggest that the actions of CGRP on gastrin release and gene expression are indirect and mediated through the paracrine influences of somatostatin.

Mechanisms of proton-induced stimulation of CGRP release from rat antrum.

Mechanisms of acid-evoked CGRP release from gastric afferent nerves were investigated in rat antral mucosal/submucosal tissues. Low pH (pH 4.0, 5.0 and 6.0) stimulated antral CGRP release significantly and dose-dependently from rat antral fragments. Removal of extracellular calcium from the incubation medium resulted in significant inhibition (59%, P < 0.001) of acid (pH 4.0)-stimulated CGRP release. Conotoxin (1 x 10(-7) M), the selective blocker of N-type calcium channels, also significantly inhibited proton (pH 4.0)-induced CGRP release to values that were 74% below net stimulated levels. Neither nifedipine (1 x 10(-6) M), the L-type Ca(2+)-channel antagonist, nor indomethacin (1 x 10(-5) M), inhibitor of prostaglandin synthesis, altered acid-induced CGRP release. In contrast, ruthenium red (1 x 10(-5) M), capsaicin antagonist, almost completely prevented acid (pH 4.0)-stimulated CGRP release. Capsazepine (1 x 10(-4) M), a specific capsaicin receptor antagonist, also completely abolished acid-induced CGRP release. In conclusion, the results of these studies indicate that hydrogen ions are capable of evoking CGRP release from peripheral sensory neurons in rat antral mucosal/submucosal tissues. Proton-evoked CGRP release requires extracellular calcium and involves N-type calcium channels. Furthermore, acid appears to exert a capsaicin-like effect to evoke sensory neuropeptide release that is sensitive to capsazepine and ruthenium red. These data suggest that proton-induced antral CGRP release represents a direct action of hydrogen ions on mucosal/submucosal sensory dendritic nerve endings to effect local release of neuropeptide.