Receptors involved in helodermin action on rat pancreatic acini

1986 ◽  
Vol 251 (5) ◽  
pp. G602-G610 ◽  
Author(s):  
J. P. Dehaye ◽  
J. Winand ◽  
C. Damien ◽  
F. Gomez ◽  
P. Poloczek ◽  
...  
Keyword(s):  
Protein Phosphorylation ◽  
Amylase Secretion ◽  
High Concentration ◽  
Amylase Release ◽  
Pancreatic Acini ◽  
Heloderma Suspectum ◽  
Low Concentrations ◽  
High Concentrations ◽  
Camp Levels ◽  
Camp Elevation

Helodermin is a new peptide isolated from the venom of Heloderma suspectum. Its effects on rat pancreatic acini were compared with those of secretin and vasoactive intestinal peptide (VIP). Four classes of receptors with decreasing affinity for secretin (S1, S2, S3, and S4) were first delineated. Occupancy of S1 and S2 by secretin was responsible for a biphasic adenosine 3',5'-cyclic monophosphate (cAMP) response. S3 was VIP preferring so that the VIP-induced increase in cAMP could be inhibited by VIP-(10 --28). S2 and S3 allowed cAMP elevation, protein phosphorylation, weak secretory effects, and potentiation of cholecystokinin octapeptide (CCK-8) when occupied by secretin and VIP, respectively. A more efficient exocytosis was observed with secretin interacting with low-affinity receptors S4. Helodermin increased cAMP levels 14-fold, this increase being inhibited by VIP-(10–28). Low concentrations of helodermin stimulated amylase secretion twofold and potentiated secretion by CCK-8. High concentrations of helodermin stimulated secretion another 2.6-fold. Helodermin bound to the four secretin receptors with a weak selectivity. At low concentration, helodermin stimulated cAMP elevation, protein phosphorylation, amylase release, and potentiation of CCK-8 through S3, whereas at high concentration it stimulated secretion via S4.

Intracellular mediators of bombesin action on rat pancreatic acinar cells

1991 ◽  
Vol 260 (6) ◽  
pp. G858-G864 ◽  
Author(s):  
T. Matozaki ◽  
W. Y. Zhu ◽  
Y. Tsunoda ◽  
B. Goke ◽  
J. A. Williams
Keyword(s):  
Time Course ◽  
Pancreatic Acinar Cells ◽  
Amylase Secretion ◽  
Inhibitory Effects ◽  
Pancreatic Acini ◽  
Low Concentrations ◽  
Intracellular Messengers ◽  
Early Peak ◽  
High Concentrations ◽  
Intracellular Mediators

The effects of bombesin on physiological responses (amylase secretion, protein synthesis) and intracellular mediators [inositol 1,4,5-trisphosphate (1,4,5–IP3), [Ca2+]i, and diacylglycerol] were studied in isolated rat pancreatic acini and compared with the actions of cholecystokinin (CCK). Bombesin stimulated amylase secretion to the same extent as CCK. However, it failed to reproduce the inhibition of amylase secretion by high concentrations of CCK and likewise did not inhibit incorporation of [3H]leucine into protein in contrast to high concentrations of CCK. Low concentrations of bombesin (1–100 pM) induced repetitive oscillations in [Ca2+]i, whereas higher concentrations of bombesin (1–10 nM) induced a large transient increase in [Ca2+]i followed by a small sustained plateau. Bombesin (1–100 nM) induced an early peak of 1,4,5–IP3 at 5–15 s but was without measurable effect at lower concentrations. These effects on [Ca2+]i and 1,4,5–IP3 were similar to those seen with CCK except that bombesin was approximately 10-fold less potent than CCK. Bombesin induced an increase in acinar 1,2-diacylglycerol with a biphasic time course similar to CCK. However, the magnitude of the response to bombesin was much smaller than the response to CCK. The results suggest that bombesin receptors initiate similar intracellular messengers as does CCK. However, CCK induces a larger increase of diacylglycerol and probably an as yet unidentified messenger responsible for its inhibitory effects.

Action of theophylline on secretagogue-stimulated amylase release from dispersed pancreatic acini

1980 ◽  
Vol 239 (4) ◽  
pp. G324-G333
Author(s):  
L. Y. Korman ◽  
M. D. Walker ◽  
J. D. Gardner
Keyword(s):  
Guinea Pig ◽  
Vasoactive Intestinal Peptide ◽  
Enzyme Secretion ◽  
Amylase Release ◽  
Pancreatic Acini ◽  
Modes Of Action ◽  
Low Concentrations ◽  
High Concentrations ◽  
Stimulation Of

In dispersed acini from guinea pig pancreas, theophylline did not alter basal amylase release, but had three functionally distinct modes of action on the stimulation of amylase release caused by various secretagogues. 1) At relatively low concentrations (0.1-1.0 mM), theophylline augmented the increase in enzyme secretion caused by vasoactive intestinal peptide, secretin, or 8-bromoadenosine 3',5'-monophosphate, but did not alter the increase in amylase release caused by other secretagogues. 2) At intermediate concentrations (1-10 mM), theophylline selectively altered the increase in enzyme secretion caused by carbamylcholine, but did not alter the effects of cholecystokinin or bombesin, secretagogues whose modes of action are similar to that of cabamylcholine. 3) At high concentrations (greater than 10 mM), theophylline inhibited the increase in enzyme secretion caused by all secretagogues tested.

Calcium dependence of proteinase-activated receptor 2 and cholecystokinin- mediated amylase secretion from pancreatic acini

2005 ◽  
Vol 289 (4) ◽  
pp. G686-G695 ◽  
Author(s):  
Anupriya Sharma ◽  
Xiaohong Tao ◽  
Arun Gopal ◽  
Brooke Ligon ◽  
Michael L. Steer ◽  
...  
Keyword(s):  
Phospholipase C ◽  
Digestive Enzymes ◽  
Extracellular Calcium ◽  
Amylase Secretion ◽  
Free Medium ◽  
Pancreatic Acini ◽  
Low Concentrations ◽  
Different Types ◽  
High Concentrations ◽  
Compare And Contrast

Pancreatic acini secrete digestive enzymes in response to a variety of secretagogues including CCK and agonists acting via proteinase-activated receptor-2 (PAR2). We employed the CCK analog caerulein and the PAR2-activating peptide SLIGRL-NH2 to compare and contrast Ca2+ changes and amylase secretion triggered by CCK receptor and PAR2 stimulation. We found that secretion stimulated by both agonists is dependent on a rise in cytoplasmic Ca2+ concentration ([Ca2+]i) and that this rise in [Ca2+]i reflects both the release of Ca2+ from intracellular stores and accelerated Ca2+ influx. Both agonists, at low concentrations, elicit oscillatory [Ca2+]i changes, and both trigger a peak plateau [Ca2+]i change at high concentrations. Although the two agonists elicit similar rates of amylase secretion, the rise in [Ca2+]i elicited by caerulein is greater than that elicited by SLIGRL-NH2. In Ca2+-free medium, the rise in [Ca2+]i elicited by SLIGRL-NH2 is prevented by the prior addition of a supramaximally stimulating concentration of caerulein, but the reverse is not true; the rise elicited by caerulein is neither prevented nor reduced by prior addition of SLIGRL-NH2. Both the oscillatory and the peak plateau [Ca2+]i changes that follow PAR2 stimulation are prevented by the phospholipase C (PLC) inhibitor U73122, but U73122 prevents only the oscillatory [Ca2+]i changes triggered by caerulein. We conclude that 1) both PAR2 and CCK stimulation trigger amylase secretion that is dependent on a rise in [Ca2+]i and that [Ca2+]i rise reflects release of calcium from intracellular stores as well as accelerated influx of extracellular calcium; 2) PLC mediates both the oscillatory and the peak plateau rise in [Ca2+]i elicited by PAR2 but only the oscillatory rise in [Ca2+]i elicited by CCK stimulation; and 3) the rate of amylase secretion elicited by agonists acting via different types of receptors may not correlate with the magnitude of the [Ca2+]i rise triggered by those different types of secretagogue.

Glutathione depletion inhibits amylase release in guinea pig pancreatic acini

1983 ◽  
Vol 244 (3) ◽  
pp. G273-G277
Author(s):  
W. F. Stenson ◽  
E. Lobos ◽  
H. J. Wedner
Keyword(s):  
Guinea Pig ◽  
Reduced Glutathione ◽  
Glutathione Depletion ◽  
Amylase Secretion ◽  
Ionophore A23187 ◽  
Amylase Release ◽  
Pancreatic Acini ◽  
Stimulus Secretion Coupling ◽  
Dose Dependent ◽  
Higher Doses

Isolated guinea pig pancreatic acini were specifically depleted of glutathione by treatment with 2-cyclohexene-1-one (2-CHX-1). Untreated acini contained 4.3 +/- 0.6 micrograms of glutathione per milligram protein. Incubation with 1 mM 2-CHX-1 for 5 min at 37 degrees C depleted glutathione to 17% of control values; 5 mM 2-CHX-1 depleted glutathione to less than 4% of control values. Incubation with 2-CHX-1 also impaired the ability of the isolated acini to secrete amylase in response to stimulation with carbachol and the ionophore A23187. The depletion of glutathione and the inhibition of amylase secretion by 2-CHX-1 were both dose dependent and time dependent. Incubation of acini with 2 mM 2-CHX-1 for 15 min at 37 degrees C reduced glutathione levels to 6.6% of control and reduced carbachol-stimulated amylase release to 63% of control. Higher doses of 2-CHX-1 or longer incubations resulted in greater depletion of glutathione and greater inhibition of carbachol-induced amylase release. These data indicate that specific depletion of glutathione impairs the ability of isolated acini to secrete amylase in response to physiological and pharmacologic stimuli and suggest that glutathione has a role in stimulus-secretion coupling in the exocrine pancreas.

scholarly journals Inhibitory effect of high leucine concentration on α-amylase secretion by pancreatic acinar cells: possible key factor of proteasome

2018 ◽  
Vol 38 (6) ◽  
Author(s):  
Long Guo ◽  
Baolong Liu ◽  
Chen Zheng ◽  
Hanxun Bai ◽  
Hao Ren ◽  
...  
Keyword(s):  
Amylase Activity ◽  
Acinar Cells ◽  
Inhibitory Effect ◽  
Proteasome Activity ◽  
Pancreatic Acinar Cells ◽  
Amylase Secretion ◽  
High Concentration ◽  
Amylase Release ◽  
Cholecystokinin Receptor ◽  
Key Factor

The present study aimed to investigate whether leucine affects the pancreatic exocrine by controlling the antisecretory factor (AF) and cholecystokinin receptor (CCKR) expression as well as the proteasome activity in pancreatic acinar cells of dairy calves. The pancreatic acinar cells were isolated from newborn Holstein bull calves and cultured using the Dulbecco’s modified Eagle’s medium/nutrient mixture F12 Ham’s liquid (DMEM/F12). There were six treatments of leucine dosage including 0 (control), 0.23, 0.45, 1.35, 4.05, and 12.15 mM, respectively. After culture for 3 h, the samples were collected for subsequent analysis. As the leucine concentration increased from 0 to 1.35 mM, the α-amylase activity in media decreased significantly (P<0.05), while further increase in leucine concentration did not show any decrease in α-amylase activity. Addition of leucine inhibited (P<0.05) the expression of AF and CCKR, and decreased the activity of proteasome (P<0.05) by 76%, 63%, 24%, 7%, and 9%, respectively. Correlation analysis results showed α-amylase secretion was negatively correlated with leucine concentration (P<0.01), and positively correlated with proteasome activity (P<0.01) and the expression of CCK1R (P<0.01) and AF (P<0.05). The biggest regression coefficient was showed between α-amylase activity and proteasome (0.7699, P<0.001). After inhibition of proteasome by MG-132, low dosage leucine decreased (P<0.05) the activity of proteasome and α-amylase, as well as the expression of CCK1R. In conclusion, we demonstrated that the high-concentration leucine induced decrease in α-amylase release was mainly by decreasing proteasome activity.

Amylase secretion by isolated pancreatic acini after acute cholecystokinin treatment in vivo

1984 ◽  
Vol 246 (4) ◽  
pp. G419-G425 ◽  
Author(s):  
M. Otsuki ◽  
Y. Okabayashi ◽  
A. Ohki ◽  
S. R. Hootman ◽  
S. Baba ◽  
...  
Keyword(s):  
Binding Sites ◽  
Amylase Secretion ◽  
Response Curves ◽  
Amylase Release ◽  
Pancreatic Acini ◽  
Cholecystokinin Receptor ◽  
Isolated Pancreatic Acini ◽  
And Control ◽  
Quinuclidinyl Benzilate

A single dose of synthetic cholecystokinin octapeptide (CCK8, 5 micrograms/kg) in a depot carrier was injected subcutaneously into rats 2 and 14 h before the removal of the pancreas and the preparation of isolated pancreatic acini. CCK8 treatment induced no significant change in body weight or total amount of pancreatic DNA, but pancreatic weight, total pancreatic protein and amylase, and the concentration of amylase and total protein relative to DNA were significantly decreased. In acini prepared from CCK8-pretreated rats, responsiveness to maximal and supramaximal concentrations of CCK8 was significantly increased, irrespective of whether the amount of amylase released was expressed relative to DNA or calculated as a percentage of the acinar content. The dose-response curves for CCK8 were similarly shaped in both CCK8-pretreated and control rats but shifted threefold toward higher concentrations of CCK8 2 or 14 h after CCK8 treatment. Specific 125I-CCK binding was significantly increased only for high-affinity binding sites. Although these observations suggest that alterations in pancreatic amylase release could be due to changes at the cholecystokinin receptor, the secretory responsiveness to maximal and supramaximal concentrations of carbachol was also increased without any change in the sensitivity. Moreover, in contrast to the cholecystokinin receptor, there was no change in the number of muscarinic receptors or in their affinity for either agonists or antagonists measured with [3H]quinuclidinyl benzilate.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Caerulein and carbamoylcholine stimulate pancreatic amylase release at resting cytosolic free Ca2+

1986 ◽  
Vol 235 (1) ◽  
pp. 139-143 ◽  
Author(s):  
R Bruzzone ◽  
T Pozzan ◽  
C B Wollheim
Keyword(s):  
Fold Increase ◽  
Free Calcium ◽  
Amylase Secretion ◽  
Rapid Phase ◽  
Secretory Rate ◽  
Amylase Release ◽  
Pancreatic Acini ◽  
Significant Stimulation ◽  
Phase 0 ◽  
Stimulation Of

Cytosolic free calcium concentrations ([Ca2+]i) and amylase secretion were measured in isolated rat pancreatic acini loaded with the intracellularly trapped fluorescent indicator quin2. Both caerulein and carbamoylcholine caused a rapid increase in [Ca2+]i, with a maximal 3-fold increase at 10(-9) M-caerulein and 10(-4) M-carbamoylcholine. However, caerulein (10(-12) M and 10(-11) M) as well as carbamoylcholine (10(-7) M) caused a significant stimulation of amylase release, while not inducing any detectable rise in [Ca2+]i. Changes in [Ca2+]i after addition of either secretagogue were transient and did not last more than 2-3 min. By contrast, when amylase secretion was monitored as a function of time, two distinct secretory phases could be observed upon addition of either carbamoylcholine (10(-5) M) or caerulein (10(-10) M). An initial, rapid phase (0-5 min) which caused a 6-7-fold increase above basal, followed by a sustained (5-30 min), but less marked, secretory rate (2-3-fold above basal). Addition of atropine (10(-4) M) 5 min after carbamoylcholine (10(-5) M) (i.e. after termination of the rise in [Ca2+]i and of the first secretory phase) did not cause any significant change in [Ca2+]i, while significantly inhibiting amylase secretion from 5 to 30 min to the same rate observed in the absence of the secretagogue. These results show that caerulein and carbamoylcholine, two agents thought to activate secretion mainly through mobilization of Ca2+ from intracellular stores, are capable of eliciting amylase secretion independently of a concomitant rise in [Ca2+]i. Furthermore, with both secretagogues the rise in [Ca2+]i, when observed, was only transient, while the stimulation of amylase release was sustained.

scholarly journals Modulation of Radula Opener Muscles in Aplysia

1999 ◽  
Vol 82 (3) ◽  
pp. 1339-1351 ◽  
Author(s):  
Colin G. Evans ◽  
Ferdinand S. Vilim ◽  
Orna Harish ◽  
Irving Kupfermann ◽  
Klaudiusz R. Weiss ◽  
...  
Keyword(s):  
Motor Neuron ◽  
Relaxation Rate ◽  
Muscle Relaxation ◽  
Muscle Fibers ◽  
Low Concentrations ◽  
Muscle Complex ◽  
Myogenic Activity ◽  
High Concentrations ◽  
Camp Levels ◽  
Rest Length

We observed fibers immunoreactive (IR) to serotonin (5-HT), the myomodulins (MMs), and FMRFamide on the I7-I10 complex in the marine mollusk Aplysia californica. The I7–I10 muscle complex, which produces radula opening, is innervated primarily by one motor neuron, B48. B48 is MM-IR and synthesizes authentic MMA. When B48 is stimulated in a physiological manner, cAMP levels are increased in opener muscles. cAMP increases also are seen when the MMs are applied to opener muscles but are not seen with application of the B48 primary neurotransmitter acetylcholine (ACh). Possible physiological sources of 5-HT and FMRFamide are discussed. When modulators are applied to resting opener muscles, changes in membrane potential are observed. Specifically, 5-HT, MMB, and low concentrations of MMA all depolarize muscle fibers. This depolarization is generally not sufficient to elicit myogenic activity in the absence of neural activity under “rest” conditions. However, if opener muscles are stretched beyond rest length, stretch- and modulator-induced depolarizations can summate and elicit contractions. This only occurs, however, if “depolarizing” modulators are applied alone. Thus other modulators (i.e., FMRFamide and high concentrations of MMA) hyperpolarize opener muscle fibers and can prevent depolarizing modulators from eliciting myogenic activity. All modulators tested affected parameters of motor neuron-elicited contractions of opener muscles. MMB and 5-HT increased contraction size over the range of concentrations tested, whereas MMA potentiated contractions when it was applied at lower concentrations but decreased contraction size at higher concentrations. FMRFamide decreased contraction size at all concentrations and did not affect relaxation rate. Additionally, the MMs and 5-HT increased muscle relaxation rate, decreased contraction latency, and decreased the rate at which tension was developed during motor neuron-elicited muscle contractions. Thus these modulators dramatically affect the ability of opener muscles to follow activity in the opener motor neuron B48. The possible physiological significance of these findings is discussed.

New proglumide-analogue CCK receptor antagonists: very potent and selective for peripheral tissues

1986 ◽  
Vol 250 (6) ◽  
pp. G856-G860 ◽  
Author(s):  
C. Niederau ◽  
M. Niederau ◽  
J. A. Williams ◽  
J. H. Grendell
Keyword(s):  
Agonist Activity ◽  
Receptor Antagonists ◽  
Maximal Inhibition ◽  
Amylase Release ◽  
Peripheral Tissues ◽  
Pancreatic Acini ◽  
Cholecystokinin Receptor ◽  
Low Concentrations ◽  
Competitive Kinetics ◽  
Cck Receptor Antagonists

The present study evaluates the ability of two recently synthesized analogues of proglumide, both 4-benzamido-N,N-di-alkyl-glutaramic acid derivatives, to act as cholecystokinin receptor antagonists. Both new antagonists inhibited cholecystokinin-stimulated amylase release and, similarly, binding of 125I-cholecystokinin to isolated rat pancreatic acini. These effects displayed competitive kinetics; both antagonists showed no agonist activity and were specific in that only those secretagogues were inhibited that interact with the cholecystokinin receptor. Both antagonists also inhibited binding of 125I-cholecystokinin to mouse pancreatic membrane particles similarly to results with rat pancreatic acini. With the more potent of the two new antagonists, half-maximal inhibition of action and binding of cholecystokinin was observed with low concentrations of approximately 10(-7) M; compared with proglumide, the new antagonists were as much as 4,000 times more potent. Unlike proglumide, which inhibits binding of cholecystokinin to pancreas and brain tissue similarly, both antagonists inhibited binding of cholecystokinin to the pancreas at much lower concentrations compared with brain. The more potent of the inhibitors was 300 times more potent in inhibiting binding of cholecystokinin to pancreatic tissues compared with brain.

A new CCK analogue differentiates two functionally distinct CCK receptors in rat and mouse pancreatic acini

1989 ◽  
Vol 257 (4) ◽  
pp. G594-G600 ◽  
Author(s):  
T. Matozaki ◽  
J. Martinez ◽  
J. A. Williams
Keyword(s):  
Competitive Inhibition ◽  
Inhibitory Effect ◽  
Leucine Incorporation ◽  
Amylase Release ◽  
Pancreatic Acini ◽  
Cck Receptors ◽  
Partial Agonists ◽  
Low Concentrations ◽  
Phenylalanine Residue ◽  
Stimulation Of

Analysis of the competitive inhibition of 125I-labeled cholecystokinin octapeptide (CCK-8) binding to isolated rat or mouse pancreatic acini showed that in both species CCK-8 interacts with two different affinity sites. A newly synthesized CCK analogue modified at the COOH-terminal phenylalanine residue totally inhibited 125I-CCK binding. This interaction occurred with sites of a single affinity in rat acini but with two different affinity sites in mouse acini. When acini were incubated with increasing concentrations of CCK-8, a biphasic stimulation of amylase release was observed. By use of rat acini, the analogs stimulated amylase release but caused no inhibition at supramaximal concentrations. By contrast, in mouse pancreatic acini, analogues showed a biphasic stimulation of amylase release similar to CCK-8. Both CCK-8 and the analogue stimulated [3H]leucine incorporation into protein at low concentrations in rat pancreatic acini. Higher concentrations of CCK-8 profoundly inhibited [3H]leucine incorporation, whereas the analogue had no inhibitory effect. Moreover, the analogue at higher concentrations blocked the inhibition of [3H]leucine incorporation caused by CCK-8 but did not affect carbamylcholine-induced inhibition. In mouse acini, however, the CCK analogue inhibited [3H]leucine incorporation similar to the effect of CCK-8. The results support the concept that occupancy of distinct affinity sites or states of the CCK receptor is associated with specific biological actions. A model of the CCK receptor is proposed in which two interchangeable affinity states exist. By occupying all the receptors in only one state, the new CCK analogues serve as partial agonists of some and antagonists of other actions of CCK.

Sign in / Sign up

Export Citation Format

Share Document