Y2 receptors for peptide YY and neuropeptide Y on dispersed chief cells from guinea pig stomach

1992 ◽  
Vol 262 (4) ◽  
pp. G756-G762
Author(s):  
G. Singh ◽  
L. Singh ◽  
J. P. Raufman
Keyword(s):  
Guinea Pig ◽  
Neuropeptide Y ◽  
Peptide Yy ◽  
Camp Production ◽  
Chief Cells ◽  
High Affinity ◽  
Cell Internalization ◽  
Pepsinogen Secretion ◽  
Guinea Pig Stomach ◽  
Dissociation Constant Kd

Peptide YY (PYY) and neuropeptide Y (NPY) inhibit agonist-induced adenosine 3',5'-cyclic monophosphate (cAMP) production and pepsinogen secretion from chief cells. We used radiolabeled PYY and NPY to characterize receptors on chief cells from guinea pig stomach. Binding of 125I-labeled PYY was rapid (70% maximal within 10 min) and specific (not inhibited by secretin, vasoactive intestinal peptide, cholecystokinin, carbachol, prostaglandin E2, forskolin, or cholera toxin). Measurement of the ability of PYY to inhibit binding of 125I-PYY indicated the presence of 1.8 x 10(3) high-affinity [dissociation constant (Kd) = 1.7 nM] and 5.1 x 10(4) low-affinity (Kd = 83.3 nM) sites/cell. Internalization of bound 125I-PYY was suggested by slow and incomplete dissociation in the presence of unlabeled PYY (50% after 2 h) and was examined further by measuring residual binding after washing with acetic acid (pH 2.5), glycine (pH 10.5), or trypsin. After 30 min at 37 degrees C, internalization of radioligand was evidenced by the failure of washing with these solutions to remove 50-65% of bound radioactivity. At 4 degrees C, internalization of 125I-PYY was nearly abolished. Binding of 125I-PYY and 125I-NPY was inhibited by NPY-(13-36) but not by [Leu31,Pro34]NPY indicating that these are Y2 receptors. In guinea pig chief cells, PYY and NPY modulate cAMP-mediated pepsinogen secretion by interacting with specific high-affinity Y2 receptors.

Actions of peptide YY and neuropeptide Y on chief cells from guinea pig stomach

1991 ◽  
Vol 260 (6) ◽  
pp. G820-G826
Author(s):  
J. P. Raufman ◽  
L. Singh
Keyword(s):  
Guinea Pig ◽  
Neuropeptide Y ◽  
G Proteins ◽  
Peptide Yy ◽  
Chief Cell ◽  
Inhibitory Effects ◽  
Camp Content ◽  
Chief Cells ◽  
Pepsinogen Secretion ◽  
Guinea Pig Stomach

Peptide YY (PYY), found in intestinal endocrine cells, and neuropeptide Y (NPY), a structural analogue of PYY found in neurons, inhibit gastric, pancreatic, and intestinal fluid and electrolyte secretion. We examined the effects of these peptides on dispersed chief cells from guinea pig stomach. PYY and NPY, but not pancreatic polypeptide, starting at nanomolar concentrations, caused a 40–50% inhibition of secretin-, vasoactive intestinal polypeptide-, prostaglandin E2-, and forskolin-induced increases in chief cell adenosine 3',5'-cyclic monophosphate (cAMP) content and pepsinogen secretion. These inhibitory peptides did not alter pepsinogen secretion caused by cholecystokinin, carbamylcholine, A23187, 8-bromo-cAMP, or a phorbol ester. The inhibitory effects of PYY on chief cell cAMP production occurred within 30 s, were independent of phosphodiesterase activity, and did not affect the actions of cholera toxin. However, the inhibitory effects of PYY were abolished when chief cells were preincubated with pertussis toxin, an agent that uncouples inhibitory guanine nucleotide binding (G) proteins from their receptors. In gastric chief cells, PYY and NPY attenuate the stimulatory effects of secretagogues whose actions are mediated by changes in cellular levels of cAMP. PYY-induced attenuation of chief cell adenylate cyclase activity appears to involve activation of inhibitory G proteins.

Actions of Helodermatidae venom peptides and mammalian glucagon-like peptides on gastric chief cells

1993 ◽  
Vol 265 (1) ◽  
pp. G118-G125 ◽  
Author(s):  
A. Rai ◽  
G. Singh ◽  
R. Raffaniello ◽  
J. Eng ◽  
J. P. Raufman
Keyword(s):  
Guinea Pig ◽  
Chief Cell ◽  
Chief Cells ◽  
Venom Peptides ◽  
High Affinity ◽  
Threefold Increase ◽  
Pepsinogen Secretion ◽  
Glucagon Like Peptide ◽  
Glucagon Like Peptide 1 ◽  
Guinea Pig Stomach

The actions of peptides (helospectin I, helodermin, exendin-3, exendin-4) that have been isolated from the venoms of Helodermatidae lizards were examined using dispersed chief cells from guinea pig stomach. These actions were compared with those of mammalian glucagon-like peptides, particularly truncated glucagon-like peptide 1 (TGLP-1), a peptide that shares 53% homology with exendin-4. The Helodermatidae venom peptides and TGLP-1 caused a two- to threefold increase in chief cell adenosine 3',5'-cyclic monophosphate and pepsinogen secretion. Exendin-3 and exendin-4 were 100 times more potent than helospectin I and helodermin and 10 times more potent than TGLP-1. Helospectin I and helodermin, but not exendin-4 or TGLP-1, inhibited the binding of 125I-labeled vasoactive intestinal peptide (VIP) and 125I-secretin to dispersed chief cells. The actions of exendin-3, exendin-4, and TGLP-1, but not those of helospectin I, helodermin, VIP, or secretin, were progressively inhibited by increasing concentrations of an exendin-receptor antagonist, exendin-(9-39)-NH2. These data indicate that in gastric chief cells, whereas the actions of helospectin I and helodermin are mediated by interaction with high-affinity secretin (low-affinity VIP) receptors, the actions of exendin-3, exendin-4, and TGLP-1 are mediated by interaction with exendin receptors.

Chief cells possess a receptor with high affinity for PACAP and VIP that stimulates pepsinogen release

1992 ◽  
Vol 263 (6) ◽  
pp. G901-G907 ◽  
Author(s):  
C. P. Felley ◽  
J. M. Qian ◽  
S. Mantey ◽  
T. Pradhan ◽  
R. T. Jensen
Keyword(s):  
Cell Function ◽  
Second Phase ◽  
Response Curves ◽  
Chief Cells ◽  
Amino Acid Peptide ◽  
High Affinity ◽  
Common Receptor ◽  
Pepsinogen Secretion ◽  
High Affinity Receptor ◽  
Guinea Pig Stomach

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a 38-amino acid peptide of the secretin-vasoactive intestinal peptide (VIP) family. To investigate whether PACAP alters chief cell function, we prepared isolated chief cells (> 90% pure) from guinea pig stomach. PACAP-38, PACAP-27, VIP, and secretin all caused a threefold increase in pepsinogen release. The dose-response curves of PACAP-38, PACAP-27, and VIP were biphasic, whereas with secretin it was not. The first phase comprised 40% of maximal release, and each of the three peptides (PACAP-38, PACAP-27, and VIP) were equipotent (EC50 0.1-0.3 nM). For the second phase, comprising 60% of maximal release, the relative potencies were PACAP-38 > PACAP-27 = VIP. 125I-labeled secretin, 125I-VIP, and 125I-PACAP-27 all demonstrated saturable binding to chief cells. Binding of both 125I-PACAP-27 and 125I-VIP was inhibited completely and with similar potencies by PACAP-38, PACAP-27, and VIP. Secretin had a > 500-fold lower affinity than PACAP-38 for displacing both 125I-PACAP-27 and 125I-VIP. With 125I-secretin, secretin was the most potent, and was 197 times more potent than PACAP-38, which was 6-8 times more potent than both PACAP-27 and VIP. We conclude that both PACAP-38 and PACAP-27 stimulate pepsinogen secretion from dispersed chief cells. In contrast to a number of other tissues, no evidence for a high-affinity receptor that interacted only with PACAP was found. PACAP and VIP interact with equal high affinity with a common receptor and with low affinity with the secretin receptor.

Ability of prostaglandin to reduce ethanol injury to dispersed chief cells from guinea pig stomach

1989 ◽  
Vol 256 (4) ◽  
pp. G704-G714 ◽  
Author(s):  
J. A. Cherner ◽  
L. Naik ◽  
A. Tarnawski ◽  
T. Brzozowski ◽  
J. Stachura ◽  
...  
Keyword(s):  
Lactate Dehydrogenase ◽  
Guinea Pig ◽  
Cell Injury ◽  
Protective Action ◽  
Control Cell ◽  
Ethanol Exposure ◽  
Fast Green ◽  
Chief Cells ◽  
Pepsinogen Secretion ◽  
Guinea Pig Stomach

To determine whether prostaglandin exerts a direct action on individual gastric epithelial cells that protects them from ethanol-induced injury, dispersed chief cells from guinea pig stomach were pretreated with 16,16-dimethyl-prostaglandin E2 (dmPGE2) or placebo before incubation with ethanol or control. Cell injury was assessed in terms of exclusion of Fast Green dye, release of lactate dehydrogenase, alterations of ultrastructure, and pepsinogen secretion stimulated by a variety of secretagogues. Of chief cells 60 +/- 2% were stained by Fast Green if incubated with 10% ethanol for 1 h after pretreatment with placebo, whereas only 38 +/- 1% of cells showed Fast Green staining when pretreated with 2.6 microM dmPGE2 before ethanol exposure. Similarly, 63 +/- 2% of cellular lactate dehydrogenase was released from chief cells pretreated with placebo compared with 36 +/- 4% of lactate dehydrogenase released from cells pretreated with 2.6 microM dmPGE2 (P less than 0.01). The prostaglandin's protective effect persisted throughout a 6-h incubation with ethanol. Scanning and transmission electron micrographs demonstrated disintegration of chief cells pretreated with placebo before ethanol exposure, whereas ultrastructural architecture was relatively preserved among chief cells pretreated with dmPGE2. Preincubation with 8 or 10% ethanol inhibited the subsequent stimulation of pepsinogen secretion caused by carbachol, cholecystokinin, A23187, 12-O-tetradecanoylphorbol 13-acetate, forskolin, or 8-bromoadenosine 3',5'-cyclic monophosphate. Pretreatment with dmPGE2 did not reduce the ethanol-induced inhibition of secretion stimulated by any of these secretagogues. These data indicate that dmPGE2 significantly reduces ethanol-induced damage to dispersed chief cells in terms of alterations of membrane permeability and ultrastructure but does not prevent the ethanol-induced impairment of pepsinogen secretion. These findings provide evidence that dmPGE2 exerts a direct but limited protective action on the gastric chief cell, independent of vascular, paracrine, or neural actions.

Actions of vasoactive intestinal peptide and secretin on chief cells prepared from guinea pig stomach

1986 ◽  
Vol 251 (1) ◽  
pp. G96-G102 ◽  
Author(s):  
V. E. Sutliff ◽  
J. P. Raufman ◽  
R. T. Jensen ◽  
J. D. Gardner
Keyword(s):  
Guinea Pig ◽  
Vasoactive Intestinal Peptide ◽  
Binding Sites ◽  
Response Curve ◽  
Close Correlation ◽  
The Other ◽  
Agonist Activity ◽  
Chief Cells ◽  
Pepsinogen Secretion ◽  
Guinea Pig Stomach

Vasoactive intestinal peptide and secretin increased cellular cAMP and pepsinogen secretion in dispersed chief cells from guinea pig gastric mucosa. With each peptide there was a close correlation between the dose-response curve for changes in cellular cAMP and that for changes in pepsinogen secretion. Vasoactive intestinal peptide-(10–28) and secretin-(5–27) had no agonist activity and antagonized the actions of vasoactive intestinal peptide and secretin on cellular cAMP and pepsinogen secretion. Studies of binding of 125I-vasoactive intestinal peptide and of 125I-secretin indicated that gastric chief cells possess four classes of binding sites for vasoactive intestinal peptide and secretin and that occupation of two of these classes of binding sites correlates with the abilities of vasoactive intestinal peptide and secretin to increase cellular cAMP and pepsinogen secretion. What function, if any, is mediated by occupation by the other two classes of binding sites remains to be determined.

Pepsinogen secretion from dispersed chief cells from guinea pig stomach

1984 ◽  
Vol 247 (1) ◽  
pp. G95-G104 ◽  
Author(s):  
J. P. Raufman ◽  
V. E. Sutliff ◽  
D. K. Kasbekar ◽  
R. T. Jensen ◽  
J. D. Gardner
Keyword(s):  
Guinea Pig ◽  
Incubation Temperature ◽  
Suitable Model ◽  
Cellular Mechanisms ◽  
Gastric Glands ◽  
Chief Cells ◽  
Pepsinogen Secretion ◽  
Mucosal Cells ◽  
Percoll Density Gradient ◽  
Guinea Pig Stomach

In the present study we examined the actions of various secretagogues on pepsinogen secretion from freshly dispersed chief cells prepared from guinea pig stomach. Chief cells were obtained by preparing dispersed gastric glands, subjecting the glands to mechanical disruption in the presence of EGTA, and fractionating the resulting mucosal cells on a Percoll density gradient. Chief cells constituted 90% of the final cell suspension and cell viability was 99%. In these cells, pepsinogen secretion was stimulated by agents whose actions are probably mediated by calcium: carbachol, cholecystokinin, and A23187. Pepsinogen secretion was also stimulated by agents whose actions are probably mediated by cAMP: secretin, vasoactive intestinal peptide, and 8-bromo-cAMP. Reducing the incubation temperature from 37 degrees to 4 degrees C or adding carbonyl cyanide m-chlorophenylhydrazone abolished secretagogue-induced pepsinogen secretion. These results indicate that freshly dispersed chief cells from guinea pig stomach are responsive to secretagogues and provide a suitable model for investigating cellular mechanisms of secretagogue-induced pepsinogen secretion.

Characterization of cholinergic receptors mediating pepsinogen secretion from chief cells

1989 ◽  
Vol 257 (2) ◽  
pp. G226-G234 ◽  
Author(s):  
V. E. Sutliff ◽  
S. Rattan ◽  
J. D. Gardner ◽  
R. T. Jensen
Keyword(s):  
Binding Sites ◽  
Binding Capacity ◽  
Close Correlation ◽  
Cholinergic Receptor ◽  
Cholinergic Receptors ◽  
Chief Cells ◽  
High Affinity ◽  
Receptor Sites ◽  
Pepsinogen Secretion ◽  
Guinea Pig Stomach

By use of a highly enriched preparation of chief cells (greater than 90% pure) prepared from guinea pig stomach, the cholinergic receptors regulating pepsinogen secretion were studied. Each of five different muscarinic cholinergic agonists, but not the nicotinic cholinergic agonist nicotine, stimulated pepsinogen release. Carbamylcholine-stimulated pepsinogen release was inhibited by each of seven different cholinergic receptor antagonists with relative potencies of N-methylscopolamine greater than scopolamine = 4-diphenylacetoxy-N-methylpiperidine methiodide = atropine much greater than pirenzepine greater than AF-DX-116 much greater than tetraethylammonium. Binding of [N-methyl-3H]scopolamine ([3H]NMS) was time and temperature dependent, reversible, saturable, and specific. Analysis of [3H]NMS binding demonstrated a Kd of 1.3 nM for NMS with a binding capacity of 61 fmol/mg protein or 5,920 sites/chief cell. With the agonists carbamylcholine, acetylcholine, or muscarine, the receptor population could be divided into two classes of receptor sites: a class with high affinity (Kd, 12-53 microM) representing 73% of the binding sites and a class with low affinity (Kd, 2-5 mM) representing 27% of the binding sites. Each of the antagonists had equal affinity for both receptor populations. There was a close correlation between the ability of antagonists to inhibit [3H]NMS binding and carbamylcholine-stimulated pepsinogen release. Each agonist was 29- to 63-fold more potent at stimulating pepsinogen release than interacting with high-affinity receptors and 2,000- to 11,000-fold more potent than interacting with low-affinity receptors, suggesting spare receptors. Studies with the alkylating agent, propylbenzilylcholine mustard, demonstrated that there was a receptor reserve of 50-80%.(ABSTRACT TRUNCATED AT 250 WORDS)

Functionally distinct receptors for cholecystokinin and gastrin on dispersed chief cells from guinea pig stomach

1988 ◽  
Vol 254 (2) ◽  
pp. G151-G155 ◽  
Author(s):  
J. A. Cherner ◽  
V. E. Sutliff ◽  
D. M. Grybowski ◽  
R. T. Jensen ◽  
J. D. Gardner
Keyword(s):  
Amino Acids ◽  
Guinea Pig ◽  
Receptor Antagonists ◽  
Chief Cells ◽  
Pepsinogen Secretion ◽  
Guinea Pig Stomach ◽  
Cck Receptor Antagonists ◽  
Stimulation Of

Caerulein, gastrin, and C-terminal fragments of cholecystokinin (CCK) varying in length from eight (CCK-8) to four (CCK-4) amino acids stimulate pepsinogen secretion from dispersed chief cells prepared from guinea pig stomach. C-terminal fragments of CCK containing fewer than four amino acids, even when tested at concentrations as high as 3 mM, do not stimulate pepsinogen secretion. The efficacies of gastrin and the various CCK-related peptides, coupled with the pattern of action of CCK receptor antagonists, indicate that chief cells from guinea pig stomach possess two functionally distinct classes of receptors, C-receptors and G-receptors. The C-receptors can be occupied by caerulein, CCK-8, CCK-7, des(SO3)CCK-8, or des(SO3)CCK-7, and occupation of C-receptors causes full stimulation of pepsinogen secretion. G-receptors can be occupied by gastrin I, gastrin II, CCK-6, CCK-5, or CCK-4, and occupation of G-receptors causes stimulation of pepsinogen secretion that is 60% of maximal.

Sign in / Sign up

Export Citation Format

Share Document