Differential control of somatostatin messenger RNA in rat gastric corpus and antrum. Role of acid, food, and capsaicin-sensitive afferent neurons.

Somatostatin messenger RNA in the antrum and corpus of rat stomach was quantified by Northern and slot blotting using a probe generated by the polymerase chain reaction. Fasting for 48 h enhanced the abundance of somatostatin mRNA in the pyloric antral region, but not in the acid-secreting region of the stomach. In fasted rats, somatostatin mRNA in antrum, but not corpus, was decreased by inhibition of acid secretion with omeprazole. In contrast, in rats treated with capsaicin to lesion small diameter afferents there was a significant decrease in somatostatin mRNA abundance in the corpus but not antrum. The effects of capsaicin cannot be attributed to nonspecific changes in gastric endocrine cell gene expression, since the abundance of histidine decarboxylase mRNA (which is a functionally regulated marker for a different gastric endocrine cell type) did not change with capsaicin. Gastric capsaicin-sensitive afferents are rich in calcitonin gene-related peptide, and in rats with antibodies to this peptide there was reduced corpus somatostatin mRNA. Moreover, infusion of calcitonin gene-related peptide in control rats produced a significant increase in somatostatin mRNA in the gastric corpus. The results indicate that somatostatin mRNA abundance is controlled by the gastric luminal contents and the extrinsic afferent innervation, but the relative importance of these factors differs in antrum and corpus: luminal contents are relatively more important in antrum and primary afferents using calcitonin gene-related peptide in the corpus.

The intrinsic and vagal extrinsic innervation of the rat stomach contains nitric oxide synthase.

Nitric oxide synthase (NOS) associated with the intrinsic and vagal extrinsic innervation of the rat gastric corpus was studied in both control and neonatally capsaicin-treated animals. Nerve cell bodies and fibres of both the myenteric and sub-mucosal plexi of the gastric corpus was found to contain NOS, but were distinct from those containing VIP. NOS-positive fibres were seen innervating the circular smooth muscle layer. Stain accumulated both proximal and distal to a vagal ligature. Staining of the nucleus of the solitary tract (NTS) was associated with specific sub-nuclei. Neonatal capsaicin treatment did not alter the staining in the NTS, vagus nerve or stomach. The data presented here support the idea that NO is a non-adrenergic non-cholinergic transmitter associated with gastric function.

The effect of ondansetron on gastric emptying in the conscious rat.

The 5-HT3 receptor antagonist ondansetron (GR38032F) enhanced the action of a protein-rich solution in delaying gastric emptying in the conscious gastric fistula rat, but had no effect on the emptying of isotonic or hypertonic saline, acid or FOY-305 which delays emptying by release of cholecystokinin (CCK). The specific CCK-A antagonist (L-364,718) increased gastric emptying of protein-rich meals. L364,718 also increased emptying in the presence of ondansetron. They indicate that protein-rich meals release both CCK and 5-hydroxytryptamine which act in different ways to control gastric motility.

Immunoneutralization suggests that calcitonin gene related peptide regulates gastric emptying in the rat.

The role of calcitonin gene related peptide (CGRP) in controlling gastric emptying was examined in conscious gastric fistula rats. Rats were immunized with CGRP conjugated to thyroglobulin to produce circulating antibodies for neutralization of the endogenous peptide; control rats received carrier thyroglobulin alone. In the latter acid, protein and hypertonic solutions delayed gastric emptying; in rats with CGRP antibodies the action of acid and hyperosmolal solutions, but not protein, was reversed. The data are compatible with the idea that acid and hyperosmolal solutions release CGRP at the peripheral terminals of visceral afferents in the upper gastrointestinal tract which in turn modifies motility to slow gastric emptying.

The role of calcitonin gene-related peptide in gastric mucosal protection in the rat.

The presence of circulating antibodies to calcitonin gene-related peptide (CGRP) enhanced the damaging effect of ethanol on the rat gastric mucosa. Taken together with previous experimental and morphological data the results suggest that CGRP released from the peripheral terminals of visceral afferent fibres plays a role in mediating gastric mucosal defence mechanisms.

The role of cholecystokinin in inhibition of gastric emptying by peptone in the rat.

It is clear that the intestinal hormone cholecystokinin (CCK) inhibits gastric emptying, but doubts remain about the physiological significance of this action. Evaluation of the apparently conflicting data is complicated by the fact that little is known of the duration of action of CCK-releasing meals in delaying emptying. We have studied this issue by following the emptying of the second of two successive liquid test meals instilled into the stomach in conscious gastric fistula rats. Prior administration of peptone, but not saline, delayed the emptying of subsequently administered saline and delayed still further the emptying of subsequently administered peptone. The action of isotonic peptone lasted about 10 min from the initial instillation into the stomach. Radioimmunoassay of plasma CCK indicated a significant increase 5 min after intragastric peptone, and a still further rise occurred 5 min after administration of the second of two consecutive peptone meals; 21 min after the first meal, plasma CCK had returned to basal levels. Intravenous infusion of CCK in a dose that matched the inhibition of gastric emptying caused by peptone gave plasma concentrations about 35% higher than those seen 5 min after the second of two consecutive peptone meals. It is concluded that a liquid test meal of peptone delays gastric emptying in part through release of CCK and that the response lasts 10 min or less. The relatively short duration of action of endogenous CCK released by a single protein-rich meal in the rat should be kept in mind in interpreting the significance of studies on the physiology of CCK.

Efferent pathways in the reflex control of gastric emptying in rats.

Previous studies have established that acid, hypertonic, or protein-rich liquid test meals delay gastric emptying by reflex pathways involving the extrinsic innervation of the gut. To characterize the efferent pathways involved in these reflexes, we have studied the emptying of liquid test meals in control rats and in rats after celiac ganglionectomy, pyloroplasty, and treatment with guanethidine or 6-hydroxydopamine, and in rats with circulating vasoactive intestinal polypeptide (VIP) antibodies. The results suggest that the action of hypertonic solutions on gastric emptying requires an intact celiac ganglion, that acid requires an intact pylorus, and that the action of protein-rich meals is suppressed by VIP antibodies. Sympathetic adrenergic neurons do not apparently mediate the gastric emptying of any of these solutions. The results suggest that there are at least three different reflexes by which the different components of a mixed meal might control gastric emptying. The results are also consistent with the idea that vagovagal reflexes mediate the action of protein-rich solutions on gastric emptying in rats.

Food stimulation of histidine decarboxylase messenger RNA abundance in rat gastric fundus.

1. Histidine decarboxylase in the enterochromaffin-like cells of the gastric corpus mucosa converts histidine to histamine which in turn stimulates gastric acid secretion. The control of histidine decarboxylase activity is poorly understood. We have examined how fasting and refeeding influence the abundance of the messenger RNA encoding histidine decarboxylase in the gastric corpus of the rat. 2. The polymerase chain reaction was used to generate a probe for detection of histidine decarboxylase messenger RNA in Northern and slot blots of total RNA from the gastric corpus of rats fasted for up to 48 h, or fasted and then refed. A gastrin monoclonal antibody was used to neutralize the action of endogenous gastrin. 3. Fasting progressively reduced histidine decarboxylase messenger RNA abundance by 3- to 4-fold after 48 h. Refeeding induced a rapid increase in histidine decarboxylase messenger RNA abundance which was detectable after 30 min. 4. There was a significant correlation between histidine decarboxylase messenger RNA abundance and plasma gastrin. Administration of gastrin antibody inhibited the increase in histidine decarboxylase activity after 6 h refeeding, but not after refeeding for 30 min. 5. The results suggest that histamine-mediated changes in postprandial acid secretion depend on control of histidine decarboxylase mRNA levels, and that gastrin regulates production of this enzyme in the rat over periods of a few hours.

Gastrin cell responses to acidification of the achlorhydric rat stomach.

In the rat, gastrin cells are normally exposed to the stimulatory effects of food and the inhibitory influences of acid in the gastric lumen. We have studied the effects of intragastric acid on gastrin cell function in animals in which the tonic inhibitory action of acid was removed by prior treatment with the proton pump blocker omeprazole. In fasted rats with gastric fistula treated with omeprazole, instillation of acid into the stomach produced a prompt decrease in plasma gastrin, but gastrin mRNA abundance showed a modest transient increase over a period of 2 h and thereafter no change; there was also a transient increase in tissue concentrations of the gastrin precursor progastrin that was compatible with increased gastrin synthesis. Concentrations of tissue gastrins, in general, increased after acid instillation, which can be attributed to continued synthesis in the presence of suppressed gastrin release. In rats fed ad libitum, a single dose of omeprazole (which produces achlorhydria for 24-30 h) produced an increase in plasma gastrin that peaked after 24 h and declined to control levels over the following 48 h; in contrast, gastrin mRNA abundance peaked 48 h after omeprazole before declining to control levels. The results indicate that whereas gastrin release might be promptly inhibited by intragastric acid, the changes in gastrin mRNA abundance are much slower: achlorhydria increases gastrin mRNA within 24 h, but acid takes longer to depress gastrin mRNA abundance. Over periods of a few hours, gastrin release and synthesis need not, therefore, change in parallel.

Gastric emptying in rats: role of afferent neurons and cholecystokinin.

Peptone, acid, and hyperosmolal saline delay gastric emptying in conscious gastric fistula rats. We have now studied the emptying of these solutions in animals pretreated with capsaicin to lesion small diameter primary afferents and in rats with both a gastric and duodenal cannula. In capsaicin-treated rats, hyperosmolal saline did not significantly inhibit gastric emptying, whereas the inhibitory action of acid and peptone was reversed but not abolished. In control rats, the action of peptone was inhibited by the selective cholecystokinin antagonist L364,718, but in capsaicin-treated rats, L364,718 enhanced the action of peptone in delaying gastric emptying. In rats with a duodenal cannula approximately 5 cm from the pylorus, intragastric peptone or hyperosmolal solutions only delayed emptying when the duodenal cannula was closed; in contrast, intragastric acid inhibited gastric emptying when the duodenal cannula was open or closed. The results suggest 1) that all three test meals delay emptying by mechanisms depending at least in part on afferent neurons; 2) peptone delays emptying by at least two mechanisms: one is mediated by cholecystokinin A-type receptors and afferent neurons, and the other requires neither these receptors nor small diameter afferents; and 3) acid, but not peptone or hyperosmolal saline, regulates emptying by an action localized to the stomach or proximal duodenum. The results suggest that there are several different reflex pathways by which liquid test meals act to delay gastric emptying.

Control of gastric corpus chromogranin A messenger RNA abundance in the rat.

Enterochromaffin-like cells in the corpus mucosa of the stomach produce histamine in response to gastrin; chromogranin A (CGA) is often used as a morphological marker for these cells, but its functional significance in the gastric mucosa is largely unknown. We have examined whether CGA mRNA abundance in the rat corpus is controlled by endogenous gastrin. In rats fasted for up to 48 h, there was a progressive decline in plasma gastrin and CGA mRNA; refeeding of fasted rats produced a prompt increase in plasma gastrin and an increase in CGA mRNA that was significant after 4 h. Treatment of fasted rats with omeprazole to inhibit acid secretion increased plasma gastrin and CGA mRNA levels. The increased CGA mRNA associated with omeprazole or refeeding was reversed by treatment of rats with the gastrin/cholecystokinin B antagonist CI-988 and gastrin antibody, respectively. The results suggest that CGA production in enterochromaffin-like cells of the rat stomach is part of the functional response of these cells to circulating gastrin.

Ranitidine bismuth citrate and ranitidine do not affect gastric emptying of a radio-labelled liquid meal.

Ranitidine bismuth citrate, a new chemical entity which is a salt complex of ranitidine and bismuth citrate, is being developed for the treatment of relapse of benign gastric and duodenal ulcer and eradication of Helicobacter pylori. The aim of the present study was to establish whether ranitidine bismuth citrate (800 mg) or ranitidine hydrochloride (300 mg) have any effect on gastric emptying of a liquid meal using gamma scintigraphy. On three separate occasions, each of twelve subjects received a single oral tablet of 800 mg ranitidine bismuth citrate, 300 mg ranitidine hydrochloride or placebo in random order. Thirty minutes after dosing each subject was given 375 ml of 99mTc-DTPA (diethylene triaminepentaacetic acid) labelled Clinifeed-ISO. The primary endpoint was the time to 50% gastric emptying (t50%). The proportion of the meal remaining was summarised by weighted mean proportion of the meal remaining in the stomach over 0-60 min and 0-180 min, separately. No differences were observed for t50%, weighted mean 0-60 min, and weighted mean 0-180 min between any two treatments. In man, we have detected no significant effect of single oral doses of ranitidine bismuth citrate 800 mg or ranitidine hydrochloride 300 mg on the rate of gastric emptying of a liquid meal when compared with placebo.