Biomedical subjects

G H Greeley

Publications and source records attributed to G H Greeley.

At least 37 records · Page 2Linked to original sources

PubMed1998

Bioactivity of intraduodenally and intravenously infused fragments of luminal cholecystokinin releasing factor (LCRF).

A luminal cholecystokinin releasing factor (LCRF), has been purified from intestinal secretion and found to have a mass of 8136 daltons. The amino-terminal 41 residues have been sequenced. Previous studies showed that intraduodenal infusion of the synthetic amino-terminal 35 amino acid peptide, LCRF1-35 significantly stimulated pancreatic protein and fluid secretion in conscious rats, but the peptide did not stimulate amylase release from isolated, dispersed pancreatic acini. In the present study, several fragments of LCRF were synthesized and tested for CCK-releasing activity (pancreatic protein secretion) to determine whether shorter fragments of LCRF exhibit the characteristic biological activity of native LCRF and synthetic LCRF1-35. Compounds tested were LCRF1-41, LCRF1-35, LCRF1-65 and LCRF11-25. Of the fragments shorter than LCRF1-35, only LCRF11-25 but not LCRF1-6 had significant CCK releasing activity. LCRF1-41 was equivalent to LCRF1-35 in potency and efficacy. Intravenous and intraduodenal infusion of LCRF1-35 elicited nearly identical dose-response curves.

Animals↗

PubMed1998

Novel expression and regulation of gastrin gene in human ovarian cancer cell line, SW626.

Gastrin-secreting tumors have been identified in ectopic locations including the ovary; the mechanisms regulating gastrin gene expression, its distribution, and signaling pathways in these ectopic tissues are not known. The purpose of our present study was to determine: (1) whether the gastrin gene and peptide could be detected in ovarian cancer cell lines, (2) if functional gastrin releasing peptide receptors (GRP-R) are present, and (3) whether gastrin gene expression is altered by GRP. Five ovarian cancer cell lines (SW626, OVCA 420, OVCA 429, OVCA 432, and OVCA 433) were analyzed. We identified gastrin gene and peptide expression in the SW626 cell line but not in the OVCA lines. SW626 cells express a functional GRP-R that is correctly coupled to the Ca2+ signaling pathway. Treatment of SW626 cells with bombesin, the amphibian equivalent of GRP, inhibited expression of the gastrin gene in a time- and dose-dependent fashion. The SW626 ovarian cancer cell line will provide a useful model to further define regulation and expression of both the gastrin gene and peptide in ectopic (nongastrointestinal) tissues.

Blotting, Northern↗

PubMed1997

Interaction of nicotine and a H2-receptor antagonist, famotidine, on gastrin and chromogranin A expression.

The purpose of this study is to examine the effect of nicotine on famotidine-induced hypergastrinemia in the rat. In addition, the effects of nicotine on gene expression for gastrin and chromogranin A (CGA) in the stomach were examined. Famotidine treatment alone (20 mg/kg. 2 x/day for 14 days) increased serum gastrin levels significantly (P < 0.05) but not antral levels of gastrin mRNA and peptide. Nicotine treatment (12 mg/kg/d) alone did not affect serum gastrin levels; however, nicotine potentiated the hypergastrinemic action of famotidine. The hypergastrinemic action of nicotine was not mediated by a downregulation of stomach somatostatin (SRIF) since stomach SRIF mRNA levels were unaffected by nicotine treatment. Administration of nicotine and famotidine also upregulated stomach CGA gene expression (i.e., mRNA and protein levels) significantly.

Animals↗

PubMed1997

Caloric restriction causes secretagogue specific changes of gastric acid secretion in rats.

The purpose of this study was to examine the effects of short-term caloric restriction (CR) for 4, 8 and 16 weeks on gastric acid secretion in rats. CR rats fed 60% of normal food intake for 4, 8 or 16 weeks and then prepared with gastric fistulas. Histamine- and carbachol-stimulated gastric acid secretion were significantly (P < 0.05) decreased after more than 4 weeks and 8 weeks of caloric restriction, respectively. In contrast, gastrin-stimulated acid secretion was unaffected by CR. The 1-h-integrated acid output to a submaximal dose of gastrin (40 micrograms.kg-1) was significantly higher than that of histamine (5 mg.kg-1) after 8 weeks of CR (63 +/- 13 and 27 +/- 4 microEq.h-1, respectively). Gastrin treatment (5 micrograms.kg-1.h-1) of CR rats restored the gastric acid responses to both histamine and carbachol. These results suggest that CR can selectively decrease the gastric acid responses to both histamine and carbachol by depletion of the endogenous tissue stores of gastrin. More importantly, these results indicate that under an in vivo gastrin-diminished condition, histamine is not the final secretagogue for gastric acid secretion.

Animals↗

PubMed1997

Distribution of prohormone convertase-6 expression in the gastrointestinal tract and effects of a fasting-refeeding regimen and a high-fat diet on ileal prohormone convertase-6 expression.

Proforms of gastrointestinal peptide hormones and neuropeptides are processed, in part, by prohormone convertases (PCs) to mature, biologically active peptides. The purpose of this study was to characterize the mRNA levels of PC-6 isoforms, PC-6A and PC-6B, in the gastrointestinal tract and pancreas of the rat, and to investigate the effects of a fasting and refeeding regimen, and a high-fat diet on ileal PC-6A expression. PC-6A mRNA is expressed throughout the entire gastrointestinal tract with the highest levels in the small intestine. Multiple-sized transcripts are present. PC-6B mRNA is expressed in the antrum and fundus of the stomach, in the small intestine, and colon. Ileal PC-6A mRNA expression increases significantly with fasting and then declines with refeeding toward control levels. Increased dietary fat increases PC-6A mRNA levels in the ileum. Since PC-6 is found throughout the entire gastrointestinal tract, it is likely that PC-6 participates in the processing of proforms of gastrointestinal peptides. The two isoforms of PC-6 have different patterns of distribution in the gastrointestinal tract and pancreas, suggesting that they process proforms of different gut peptides.

Animals↗

PubMed1997

Distribution and localization of a novel cholecystokinin-releasing factor in the rat gastrointestinal tract.

The purpose of this study was to examine the distribution and localization of an intestinal cholecystokinin (CCK)-releasing factor, called luminal CCK-releasing factor (LCRF), in the gastrointestinal tract and pancreas of the rat. RIA analysis indicates that LCRF immunoreactivity is found throughout the gut including the pancreas, stomach, duodenum, jejunum, ileum, and colon with the highest levels in the small intestine. Immunohistochemistry analysis shows LCRF immunoreactivity staining in intestinal villi, Brunner's glands of the duodenum, the duodenal myenteric plexus, gastric pits, pancreatic ductules, and pancreatic islets. These results indicate potential sources for secretagogue-stimulated release of luminal LCRF and support the hypothesis that LCRF is secreted into the intestinal lumen to stimulate CCK release from mucosal CCK cells.

Animals↗

PubMed1997

Effect of carboxypeptidase E deficiency on progastrin processing and gastrin messenger ribonucleic acid expression in mice with the fat mutation.

Proforms of gastrointestinal peptides are cleaved at paired basic residues into intermediate forms. Paired basic residues at the C-terminal then are excised by carboxypeptidases before the peptide is amidated. An obese mouse, called Cpe(fat)/Cpe(fat), has a missense mutation in carboxypeptidase E (CPE) with no pancreatic CPE activity and a reduced processing of pancreatic proinsulin to insulin. The purpose of this study was 1) to look for the presence of CPE in the antrum of the stomach, duodenum, and colon in the Cpe(fat)/Cpe(fat) mouse; 2) to determine whether CPE is involved in the processing of progastrin (Pro-G) to its carboxyl-terminal amidated form; and 3) to determine whether a decrease in amidated gastrin results in an up-regulation of stomach gastrin messenger RNA (mRNA) levels. In Cpe(fat)/Cpe(fat) mice, CPE activity was absent in the antrum and colon. In Cpe(fat)/Cpe(fat) mice, amidated gastrin levels were reduced significantly. Levels of the precursor for amidated gastrin (gastrin-Gly-Arg-Arg) were markedly elevated. Gastrin mRNA levels were increased approximately 2-fold over the levels in Cpe(fat)/Cpe(fat) mice. These results indicate that CPE is needed for processing progastrin to gastrin in the stomach and that amidated gastrin exerts an inhibitory feedback effect on gastrin mRNA levels.

Animals↗

PubMed1997

Comparison of somatostatin and pancreastatin on secretion of gastrin, pancreatic polypeptide, and peptide YY.

The purpose of this study was to compare the effects of pancreastatin (PST) (400 pmol/kg/hr) and somatostatin (SRIF) (400 pmol/kg/hr) on food-induced release of gastrin, pancreatic polypeptide (PP), and peptide YY (PYY) in conscious dogs. The present findings indicate that SRIF is more potent than PST on the inhibition of food-induced release of PP; that SRIF and PST do not influence food-induced release of gastrin; and that PST cannot inhibit food-induced release of PYY, whereas SRIF inhibits PYY release in a potent fashion.

Animals↗

PubMed1996

Influence of nicotine on gastrin and peptide YY in the rat.

The objective of this study was to examine the effects of nicotine and high-fat diets on gastrin and peptide YY (PYY) homeostasis in the rat. Antral levels of gastrin mRNA and peptide and ileal and colonic levels of PYY mRNA and peptide were examined. Serum levels of gastrin in response to food were also measured. Control rats were ad-lib fed or pair-fed according to the daily food intake of nicotine-treated rats. The results of this study indicate that nicotine treatment and fat diets can influence gastrin and PYY gene expression in the gastrointestinal tract.

Animals↗

PubMed1996

Short-term caloric restriction augments age-related decreases in gastrin content and release.

Aging is associated with significant structural and functional changes in the gastrointestinal tract. Gastrin, a hormone produced by G cells in the antrum of the stomach, stimulates proliferation of gastric mucosa; its synthesis appears to decrease with age. Life-long restriction of caloric intake is the only experimental manipulation that has been shown to retard aging processes in rats. The purpose of this study was to examine the effect of short-term caloric restriction (CR) on the production and release of the hormone gastrin with aging. Aging causes a fall in both fasting plasma levels of gastrin and antral content of gastrin in Fischer 344 rats; short-term CR appears to augment this age-related decrease. Steady state levels of antral gastrin mRNA were decreased with aging, and short-term CR resulted in an augmented decrease in aged, but not in young rats. Our findings indicate that gastrin release, synthesis and gene expression decrease with age. Restriction of the caloric intake for a short period (i.e. 8 weeks) augments this age-related decrease in antral gastrin and fasting plasma levels. Short-term CR appears to decrease the production of gastrin at the level of gene expression.

Aging↗

PubMed1996

Growth hormone upregulates gastrin and peptide YY gene expression.

The purpose of these studies was to examine the effects of excess growth hormone (GH) on gastrin and peptide YY (PYY) gene expression. Transgenic mice with the bovine GH gene linked to a mouse metallothionein I promoter were used as a model of chronic GH excess. Antral gastrin mRNA and peptide levels were elevated significantly (P < 0.05) in GH transgenic mice compared with wild type littermates. Ileal PYY mRNA and ileal and colonic PYY levels were significantly elevated in GH transgenic mice compared with wild type littermates. The elevations in gastrin and PYY gene expression in GH transgenic mice were independent of food intake. Serum concentrations of gastrin and PYY were also elevated in GH transgenic mice. Immunohistochemical analysis showed that the density of PYY-containing cells in the colon of GH transgenic mice and wild type littermates did not differ. In addition, the mRNA and protein levels of chromogranin A, a marker of endocrine cells, were not increased in the colon of GH transgenic mice. Together, these data indicate that GH, insulin-like growth factor I, or both can upregulate gastrointestinal gastrin and PYY gene expression directly.

Animals↗

PubMed1996

Glycine-extended gastrin potentiates gastrin-stimulated gastric acid secretion in rats.

The purpose of this study was to examine whether an intermediate form of amidated gastrin, glycine-extended gastrin (Gly-G), can stimulate gastric acid secretion in conscious rats prepared with gastric fistulas. Intravenous administration of Gly-G (20 nmol.kg-1.h-1) alone for 2 h did not stimulate gastric acid secretion; however, administration of Gly-G (20 nmol.kg-1.h-1) in combination with a bolus administration of gastrin (9.5 nmol/kg) potentiated acid secretion significantly. Gastric acid secretion in response to gastrin alone and gastrin plus Gly-G (2 nmol.kg-1.h-1) was 109.1 +/- 21.6 and 170.1 +/- 27.7 mueq.kg-1.h-1, respectively (P < 0.05). Gastric acid secretion in response to gastrin alone and gastrin plus Gly-G (20 nmol.kg-1.h-1) was 84.8 +/- 17.5 and 164.1 +/- 29.3 mueq.kg-1.h-1, respectively (P < 0.05). Intravenous administration of Gly-G (20 nmol.kg-1.h-1) failed to increase histamine (1 mg/kg)-stimulated acid output. These results demonstrate that Gly-G can selectively potentiate the stimulatory effect of gastrin on acid secretion in rats and that the unprocessed form of gastrin, Gly-G, can exert a biological effect in the stomach.

Animals↗

PubMed1996

Regulation of peptide YY homeostasis by gastric acid and gastrin.

Peptide YY (PYY) is a gut hormone localized primarily in the distal bowel. Because circulating PYY inhibits gastric acid secretion, we investigated the effects of gastric acid secretion and gastrin on gene expression and secretion of PYY. In conscious dogs, PYY release in response to oral food was inhibited (P < 0.05) by pharmacologic inhibition of gastric acid secretion (omeprazole, famotidine). In rats, omeprazole treatment resulted in a significant elevation in serum gastrin concentrations and a simultaneous decrease in PYY messenger RNA (mRNA) and peptide levels in the colon; administration of a gastrin receptor antagonist (L365, 260) prevented the inhibitory actions of omeprazole on colonic PYY mRNA levels. In athymic-nude mice, implantation of a human gastrinoma resulted in an elevation of serum gastrin concentrations and a concomitant depression of colonic PYY mRNA levels. We conclude that endogenous gastric acid secretion up-regulates PYY release and PYY mRNA expression. Circulating gastrin acts to down-regulate PYY release and PYY mRNA expression. This study provides evidence that foregut functions (i.e., gastric acid secretion and gastrin release) exert control over an antiacid signal (e.g. PYY release) emanating from the hindgut.

Animals↗

PubMed1995

Downregulation of prohormone convertase-1 by a phorbol ester.

Acute TPA treatment (1h, 100nM) of a human pancreatic carcinoid cell line (BON) depletes cell contents of chromogranin A (CGA) and pancreastatin (PST), a peptide derived posttranslationally from CGA. Despite removal of TPA, BON cells continue to release CGA in an unregulated fashion whereas PST secretion is reduced substantially. TPA treatment also reduced prohormone convertase-1 (PC-1) protein and increased PC-1 mRNA levels. Together, these findings indicate that the TPA-induced switch from a regulated to unregulated pattern of CGA secretion is accompanied by a decrease in the processing of CGA to PST and a decrease in the active form of a processing enzyme potentially involved in processing CGA to a smaller peptide, PST.

Aspartic Acid Endopeptidases↗

PubMed1995

Intestinal peptide YY: ontogeny of gene expression in rat bowel and trophic actions on rat and mouse bowel.

The purpose of this study was twofold: 1) to characterize the profile of colonic peptide YY (PYY) gene expression in rats and 2) to examine for potential trophic effects of PYY on the intestine in rats and mice. Expression of PYY mRNA (analyzed by Northern blotting and in situ hybridization) and PYY (analyzed by high-performance liquid chromatography and radioimmunoassay) was detected initially at day 17 of gestation in colonic extracts of Sprague-Dawley and Fischer rats. Expression of colonic PYY mRNA increased until 7 days of age and remained at its highest level (approximately twofold greater than the adult level) through the end of the nursing period. After weaning (21 days of age), PYY mRNA levels declined quickly to adult levels. Colonic PYY concentrations followed, in a coordinated manner, with some temporal delay after birth, the increase and decrease of its mRNA. Administration of PYY increased the weight and DNA content of the duodenum significantly in nursing rats and adult mice. In mice, PYY treatment also increased weight and DNA content of the ileum and colon. The trophic effects of PYY were dose related, peptide specific, and independent of species and sex. From these findings, we hypothesize that PYY plays an important role in intestinal development and dietary adaptation.

Aging↗

PubMed1994

Age-dependent increase of neurotensin expression in the ileum.

The proliferative activity of gut mucosa is increased with aging. Neurotensin (NT), a tridecapeptide localized to the distal small bowel, stimulates growth of gut mucosa; the effects of aging on gut NT are not known. Young (4-mo-old), adult (1-yr-old), and aged (2-yr-old) male Fischer 344 rats were killed; the ileal mucosa was scraped, weighed, and extracted for measurement of NT mRNA by slot-blot and Northern hybridization; and the relative NT transcription rate was determined by a nuclear run-on assay. In addition, NT tissue content and plasma levels were determined by radioimmunoassay, and full-thickness sections of ileum were examined for age-dependent alterations of NT endocrine cell (N-cell) number using immunohistochemical staining. Slot-blot and Northern-blot analyses showed that the steady-state levels of NT mRNA were increased threefold in the adult group and eightfold in the aged rats. In addition, NT peptide content and plasma levels were significantly increased in the aged group. The dramatic increases in the abundance of NT mRNA were not associated with increases in either NT transcription or N-cell number. In summary, we have demonstrated an age-dependent increase in the constitutive levels of ileal NT mRNA that appears not to be due to concomitant increases in transcription, suggesting that NT mRNA is increased by mechanisms involving mainly a posttranscriptional process. In addition, we have shown corresponding increases in the levels of both tissue and plasma NT with aging, indicating that expression of NT does not remain the same throughout life but actually increases with aging in the rat.

Aging↗

PubMed1993

Neural regulation of peptide YY secretion.

The purpose of these experiments was to investigate the neural control of peptide YY (PYY) secretion. The effects of various pharmacological manipulations and vagotomy on peptide YY (PYY) secretion was examined in dogs. Atropine, hexamethonium and atropine plus hexamethonium treatment blocked food-induced release of PYY significantly. Integrated release of PYY in response to food alone and in combination with atropine, hexamethonium and atropine plus hexamethonium were 8.8 +/- 2.2, -1.1 +/- 2.3, -2.7 +/- 2.2 and -3.2 +/- 3.1 (ng (0-150) min/ml), respectively. beta-Adrenergic blockade with propranolol or depletion of nerve terminal stores of catecholamines with reserpine did not affect food-stimulated release of PYY. Truncal vagotomy resulted in significant elevations of basal and food-induced release of PYY. IV administration of bethanechol, a cholinergic agonist, and electrical stimulation of the vagus nerve resulted in release of PYY. Together, these data suggest that food-stimulated PYY secretion is dependent on ganglionic transmission and an atropine-blockable postganglionic parasympathetic pathway; and that PYY release is inhibited tonically, probably through a vagal cholinergic mechanism. Adrenergic pathways do not participate in food-stimulated PYY release; however, electrical stimulation of the splanchnic nerves increased basal levels of PYY, suggesting that the sympathetic nervous system affects release of PYY.

Animals↗

PubMed1993

Effects of aging on gastrin and somatostatin secretion from isolated perfused rat stomach.

We have examined the release of gastrin and somatostatin from the isolated perfused stomach of rats of three different age groups (4 months, 12 months, and 24 months old) in response to bombesin and carbachol. The basal release of gastrin was diminished in 24-month-old rats. Basal somatostatin release showed an age-related decrease. Bombesin (10(-10) and 10(-9) M) and carbachol (10(-8) and 10(-7) M) stimulated gastrin release in each age group. The integrated release of gastrin in response to bombesin (10(-10) and 10(-9) M) or carbachol (10(-8) M) did not differ among the three age groups, although integrated gastrin release in response to carbachol (10(-7) M) decreased in 24-month-old rats. Bombesin-stimulated release of somatostatin decreased in 12- and 24-month-old rats. Carbachol inhibited release of somatostatin in each age group. Compared with 4-month-old rats, the inhibition of somatostatin release by carbachol was less in 24-month-old rats at 10(-8) and 10(-7) M, and less in 12-month-old rats at 10(-7) M. The decreased basal gastrin secretion and well-preserved gastrin response were further confirmed in conscious aged rats tested by means of oral gavage with 10% peptone. Our findings indicate that gastrin response to the stimuli is well preserved with aging, whereas the response of somatostatin diminishes in an age-related manner. Aging has different effects on the release of gastrin and somatostatin from the rat stomach.

Aging↗