Mutant analysis suggests that cyclic GMP mediates the cyclic AMP-induced Ca2+ uptake in Dictyostelium

1991 ◽  
Vol 99 (1) ◽  
pp. 187-191
Author(s):  
S. Menz ◽  
J. Bumann ◽  
E. Jaworski ◽  
D. Malchow
Keyword(s):  
Cyclic Amp ◽  
Mutant Strain ◽  
Cyclic Gmp ◽  
Parental Strain ◽  
Dependent Manner ◽  
Heavy Chains ◽  
Signal Relay ◽  
Sensitive Electrode ◽  
Dose Dependent ◽  
Cyclic Amp Synthesis

Previous work has shown that streamer F (stmF) mutants of Dictyostelium discoideum exhibit prolonged chemotactic elongation in aggregation fields. The mutants carry an altered structural gene for cyclic GMP phosphodiesterase resulting in low activities of this enzyme. Chemotactic stimulation by cyclic AMP causes a rapid transient increase in the cyclic GMP concentration followed by association of myosin heavy chains with the cytoskeleton. Both events persist several times longer in stmF mutants than in the parental strain, indicating that the change in association of myosin with the cytoskeleton is transmitted directly or indirectly by cyclic GMP. We measured the cyclic AMP-induced Ca2+ uptake with a Ca(2+)-sensitive electrode and found that Ca2+ uptake was prolonged in stmF mutants but not in the parental strain. The G alpha 2 mutant strain HC33 (fgdA), devoid of InsP3 release and receptor/guanylate cyclase coupling, lacked Ca2+ uptake. However, the latter response and cyclic GMP formation were normal in the signal-relay mutant strain agip 53 where cyclic AMP-stimulated cyclic AMP synthesis is absent. LiCl, which inhibits InsP3 formation in Dictyostelium, blocked Ca2+ uptake in a dose-dependent manner. The data indicate that the receptor-mediated Ca2+ uptake depends on the InsP3 pathway and is regulated by cyclic GMP. The rate of Ca2+ uptake was correlated in time with the association of myosin with the cytoskeleton, suggesting that Ca2+ uptake is involved in the motility response of the cells.

Spike-shaped oscillations in the absence of measurable changes in cyclic AMP concentration in a mutant of Dictyostelium discoideum

1987 ◽  
Vol 87 (5) ◽  
pp. 723-730
Author(s):  
B. Wurster ◽  
R. Mohn
Keyword(s):  
Light Scattering ◽  
Cyclic Amp ◽  
Cell Surface ◽  
Dictyostelium Discoideum ◽  
Cyclic Gmp ◽  
Parent Strain ◽  
Reaction System ◽  
Cell Suspensions ◽  
Deficient Mutant ◽  
Cyclic Amp Synthesis

Periodic activities of Dictyostelium discoideum cells involve two types of oscillations, spike-shaped and sinusoidal. Spike-shaped oscillations are accompanied by the periodic synthesis and release of cyclic AMP, and cyclic AMP-activated cyclic AMP synthesis is believed to control these oscillations. Experiments described here call into question the importance of cyclic AMP in spike-shaped oscillations. Cell suspensions of strain agip43, an aggregation-deficient mutant of D. discoideum, displayed spike-shaped oscillations in light scattering with period lengths about 1.5 times larger than those of the parent strain. These oscillations were not accompanied by measurable oscillations of cyclic AMP and cyclic GMP. Applied cyclic AMP pulses elicited increases of two- to threefold in the cyclic AMP level and increases of seven- to ninefold in the cyclic GMP concentration. Cyclic AMP additions caused phase shifts in the oscillations of agip43 cells, suggesting that cyclic AMP receptors at the cell surface communicate with the oscillator. We interpret these results in terms of an oscillator not based on cyclic AMP. This oscillator should be coupled to the reaction system involving cyclic AMP synthesis and release. The latter can operate in an oscillatory manner in the parent strain Ax2 but not in mutant agip43.

Cyclic nucleotides, calcium, bicarbonate, and protein secretion in canine pancreatic juice in response to cholecystokinin–pancreozymin

1979 ◽  
Vol 57 (6) ◽  
pp. 541-546 ◽  
Author(s):  
H. L. Cailla ◽  
H. Sarles ◽  
M. V. Singer
Keyword(s):  
Cyclic Amp ◽  
Pancreatic Juice ◽  
Cyclic Gmp ◽  
Cyclic Nucleotides ◽  
Parallel Increase ◽  
Calcium Bicarbonate ◽  
Strong Linear Correlation ◽  
Protein Output ◽  
Dose Dependent ◽  
Stimulation Of

The secretion of cyclic AMP, cyclic GMP, protein, calcium, and bicarbonate in the pancreatic juice of three nonanesthetized dogs with chronic gastric and duodenal Thomas cannulae has been studied. Intravenous infusions of increasing doses of cholecystokinin–pancreozymin (CCK) (1.5, 3, 6, 12, 24 Crick Harper-Raper (CHR) U kg−1 h−1) were administered together with a continuous submaximal dose of secretin (1 clinical unit (CU) kg−1 h−1). Doubling CCK doses every 45 min induced a parallel increase in the output of both cyclic nucleotides. Cyclic AMP output peaked at between 15 and 30 min for 3 and 6 U kg−1 h−1 of CCK and later for 12 and 24 U kg−1 h−1 of CCK whereas cyclic GMP output increased more constantly. Calcium output followed a pattern similar to that of cyclic GMP secretion. Flow rate and protein output attained their peaks at between 30 and 45 min. A strong linear correlation was found between the quantities of cyclic AMP, cyclic GMP, and the quantities of protein secreted in response to each CCK dose. This study demonstrates the presence of cyclic GMP in the canine pancreatic juice and the dose-dependent stimulation of the secretion of cyclic GMP and cyclic AMP by CCK in the presence of secretin.

Mode of Action of the Hypertrehalosaemic Peptides from the American Cockroach

1985 ◽  
Vol 40 (9-10) ◽  
pp. 670-676 ◽  
Author(s):  
Gerd Gäde
Keyword(s):  
Cyclic Amp ◽  
Adenylate Cyclase ◽  
Mode Of Action ◽  
Glycogen Phosphorylase ◽  
Fat Body ◽  
American Cockroach ◽  
Dependent Manner ◽  
Low Concentrations ◽  
First Time ◽  
Dose Dependent

Abstract Although crude extracts of cockroach (Periplaneta amencana) corpora cardiaca have been shown previously to affect the activity of adenylate cyclase and phosphorylase, we demonstrate in the present study for the first time that low concentrations (0.5 to 5 pmol) of the synthetic myoactive peptides. M I and M II, also affect these systems; these myoactive peptides are identical to the hypertrehalosaemic hormones I and II, and cause an increase in the concentration of the second messenger cyclic AMP in the fat body.In addition, both octapeptides activate fat body glycogen phosphorylase and promote breakdown of fat body glycogen. Both peptides increase the levels to haemolymph carbohydrate in a dose-dependent manner.

Biochemical parameters regulating forward motility initiation in vitro in goat immature epididymal spermatozoa

1998 ◽  
Vol 10 (4) ◽  
pp. 299 ◽  
Author(s):  
Bijay S. Jaiswal ◽  
Gopal C. Majumder
Keyword(s):  
Cyclic Amp ◽  
Sperm Motility ◽  
Biochemical Parameters ◽  
Vital Role ◽  
Dependent Manner ◽  
Alkaline Ph ◽  
Heat Stable ◽  
Heat Stable Protein ◽  
Dose Dependent

An investigation was carried out to analyse the biochemical parameters influencing forward motility (FM) initiation in vitro in the goat caput-epididymal immature spermatozoa. Forward motility was induced in approximately 55% of caput-sperm upon incubation in an alkaline (pH 8.0) modified Ringer’s solution containing theophylline (30 mM) (an inhibitor of cyclic AMP phosphodiesterase), dialysed epi-didymal plasma (EP) and bicarbonate. Both EP and bicarbonate induced sperm motility in a dose-dependent manner, and at saturating doses EP (0.6 mg protein mL–1) and bicarbonate (25 mM) induced FM in approx-imately 38% and 44% of the cells, respectively. The motility-promoting efficacy of EP was attributed to a heat-stable protein termed ‘forward motility protein’ (FMP). Bicarbonate served as an initiator as well as a stabilizer of FM and its action was not dependent on FMP. FMP can induce FM in the caput-sperm, but it is not essential for sperm motility initiation. Alteration of the medium pH from 6.60 to 8.00 caused a marked increase in the EP or bicarbonate-dependent sperm FM initiation, as well as intrasperm pH. At the physio-logical pH, bicarbonate served as a much more potent motility activator than FMP, although both the motility promoters showed maximal efficacy at alkaline pH (~7.8). EP as well as bicarbonate elevated the intrasperm cyclic AMP level. Unlike EP, bicarbonate is capable of increasing intrasperm pH. The intrasperm pH increased from 6.54 0.02 to 6.77 0.03 during sperm transit from caput to cauda. The data are con-sistent with the view that FMP activates sperm forward motility by enhancing the intrasperm cyclic AMP level and that extracellular bicarbonate and pH play a vital role in the initiation of sperm FM during the epi-didymal transit.

Influence of dopamine on cyclic nucleotide enzymes in bovine retinal membrane fractions

1993 ◽  
Vol 10 (6) ◽  
pp. 991-996 ◽  
Author(s):  
Ari Sitaramayya ◽  
Lorraine Lombardi ◽  
Alexander Margulis
Keyword(s):  
Cyclic Amp ◽  
Cyclic Gmp ◽  
Cyclic Nucleotide ◽  
D2 Receptors ◽  
D1 Receptors ◽  
Metabolizing Enzymes ◽  
D2 Agonist ◽  
Hydrolysis Of ◽  
Cyclic Amp Synthesis

AbstractDopamine is a major neurotransmitter and neuromodulator in vertebrate retina. Although its pharmacological and physiological actions are well understood, the biochemical mechanisms of its signal transduction are less clear. Acting via D1 receptors, dopamine was shown to increase cyclic AMP levels in intact retina and to activate adenylate cyclase in retinal homogenates. The action via activation of D2 receptors is controversial: it was reported to decrease cyclic AMP levels in intact retina but inhibition of cyclase could not be demonstrated in retinal homogenates; also it was reported to activate rod outer segment cyclic GMP phosphodiesterase in vitro but did not decrease cyclic GMP levels in aspartate-treated retinas. We made an attempt to fractionate bovine retinal membranes and to investigate the effects of dopamine, via Dl and D2 receptors, on the synthesis and hydrolysis of cyclic AMP and cyclic GMP. Activation of cyclic AMP synthesis was noted in all fractions, but no effects were evident on cyclic nucleotide hydrolysis or cyclic GMP synthesis in any fraction. Also, D2 agonist did not inhibit cyclic AMP synthesis. These observations suggest that D2 receptors may not be directly coupled to cyclic nucleotide metabolizing enzymes in bovine retina.

scholarly journals Calcium rather than cyclic AMP is an intracellular messenger of parathyroid hormone action on glycogen metabolism in isolated rat hepatocytes

1989 ◽  
Vol 258 (3) ◽  
pp. 889-894 ◽  
Author(s):  
T Mine ◽  
I Kojima ◽  
E Ogata
Keyword(s):  
Parathyroid Hormone ◽  
Cyclic Amp ◽  
Glucose Production ◽  
Rat Hepatocytes ◽  
Free Calcium ◽  
Dependent Manner ◽  
Isolated Rat Hepatocytes ◽  
Glucose Output ◽  
Dose Dependent ◽  
Isolated Rat

The synthetic 1-34 fragment of human parathyroid hormone (1-34hPTH) stimulated glucose production in isolated rat hepatocytes. The effect of 1-34hPTH was dose-dependent and 10(10) M-1-34 hPTH elicited the maximum glucose output, which was approx. 80% of that by glucagon. Although 1-34hPTH induced a small increase in cyclic AMP production at concentrations higher than 10(-9) M, 10(-10) M-1-34hPTH induced the maximum glucose output without significant elevation of cyclic AMP. This is in contrast to the action of forskolin, which increased glucose output to the same extent as 10(-10) M-1-34hPTH by causing a 2-fold elevation of cyclic AMP. In addition to increasing cyclic AMP, 1-34hPTH caused an increase in cytoplasmic free calcium concentration ([Ca2+]c). When the effect of 1-34hPTH on [Ca2+]c was studied in aequorin-loaded cells, low concentrations of 1-34hPTH increased [Ca2+]c: the 1-34hPTH effect on [Ca2+]c was detected at as low as 10(-12) M and increased in a dose-dependent manner. 1-34hPTH increased [Ca2+]c even in the presence of 1 microM extracellular calcium, suggesting that PTH mobilizes calcium from an intracellular pool. In line with these observations, 1-34hPTH increased the production of inositol trisphosphate. These results suggest that: (1) PTH activates both cyclic AMP and calcium messenger systems and (2) PTH stimulates glycogenolysis mainly via the calcium messenger system.

scholarly journals Cross-talk between glucagon- and adenosine-mediated signalling systems in rat hepatocytes: effects on cyclic AMP-phosphodiesterase activity

1995 ◽  
Vol 312 (3) ◽  
pp. 763-767 ◽  
Author(s):  
M Robles-Flores ◽  
G Allende ◽  
E Piña ◽  
J A García-Sáinz
Keyword(s):  
Cyclic Amp ◽  
Pertussis Toxin ◽  
Plasma Membranes ◽  
Rat Hepatocytes ◽  
Dependent Manner ◽  
Phosphodiesterase Activity ◽  
Cyclic Amp Phosphodiesterase ◽  
Cyclic Amp Accumulation ◽  
A1 Adenosine Receptors ◽  
Dose Dependent

The effect of adenosine analogues on glucagon-stimulated cyclic AMP accumulation in rat hepatocytes was explored. N6-Cyclopentyladenosine (CPA), 5′-N-ethylcarboxamidoadenosine and N6-(R-phenylisopropyl)adenosine inhibited in a dose-dependent manner the cyclic AMP accumulation induced by glucagon. This effect seems to be mediated through A1 adenosine receptors. Pertussis toxin completely abolished the effect of CPA on glucagon-stimulated cyclic AMP accumulation in whole cells which suggested that a pertussis-toxin-sensitive G-protein was involved. On the other hand, this action of adenosine analogues on glucagon-induced cyclic AMP accumulation was reverted by the selective low-Km cyclic AMP-phosphodiesterase inhibitor Ro 20-1724. Analysis of cyclic AMP-phosphodiesterase activity in purified hepatocyte plasma membranes showed that glucagon in the presence of GTP inhibited basal PDE activity by 45% and that CPA reverted this inhibition in dose-dependent manner. In membranes derived from pertussis-toxin-treated rats, we observed no inhibition of cyclic AMP-phosphodiesterase activity by glucagon in the absence or presence of CPA. Our results indicate that in hepatocyte plasma membranes, stimulation of adenylate cyclase activity and inhibition of a low-Km cyclic AMP phosphodiesterase activity are co-ordinately regulated by glucagon, and that A1 adenosine receptors can inhibit glucagon-stimulated cyclic AMP accumulation by blocking glucagon's effect on phosphodiesterase activity.

scholarly journals Changes in adenosine receptors during differentiation of 3T3-F442A cells to adipocytes

1988 ◽  
Vol 249 (2) ◽  
pp. 377-381 ◽  
Author(s):  
K Ravid ◽  
J M Lowenstein
Keyword(s):  
Cyclic Amp ◽  
Adenylate Cyclase ◽  
Adenosine Receptors ◽  
Positive Response ◽  
Inhibitory Effect ◽  
Dependent Manner ◽  
Similar Extent ◽  
Phenylisopropyl Adenosine ◽  
Dose Dependent ◽  
Basal Concentration

Incubation of undifferentiated 3T3-F442A cells (preadipocytes) with 5′-N-ethylcarboxamidoadenosine (NECA) increases intracellular cyclic AMP in a dose-dependent manner. The effect of NECA is antagonized by 8-phenyltheophylline, but potentiated by 4-(3-butoxy-4-methoxybenzyl)-2-imidazolidine, an inhibitor of cyclic AMP phosphodiesterase. Incubation of preadipocytes with (-)-N6-(R-phenylisopropyl)adenosine (PIA) has no inhibitory effect on the basal concentration of cyclic AMP or on the stimulation of adenylate cyclase by isoprenaline or forskolin. Micromolar concentrations of PIA increase intracellular cyclic AMP, but with a lower potency than NECA. Similar findings are obtained with the non-differentiating cell line 3T3-C2. Thus preadipocyte 3T3-F442A cells and 3T3-C2 cells appear to express only stimulatory adenosine receptors. For some time after 3T3-F442A cells have differentiated to adipocytes, micromolar concentrations of NECA and PIA continue to increase cyclic AMP to a similar extent to that in preadipocytes, whereas nanomolar concentrations of PIA decrease the stimulatory effects of isoprenaline and forskolin on adenylate cyclase by 50%. However, several days after differentiation, the adipocytes gradually lose the major part of their positive response to NECA and reach a steady response to NECA 10 days after differentiation. The inhibition of adenylate cyclase caused by PIA remains constant for at least 2 weeks after differentiation. With membranes derived from the cells, the effects of NECA and PIA depend on GTP. These results indicate that, during the differentiation of 3T3-F442A cells to adipocytes, new inhibitory adenosine receptors are expressed, whereas the stimulatory receptors become attenuated.

CYCLIC AMP AND CYCLIC GMP ACCUMULATION BY ISOLATED HAMSTER GRANULOSA CELLS STIMULATED BY LH AND FSH

1978 ◽  
Vol 89 (1) ◽  
pp. 173-181 ◽  
Author(s):  
Anastasia Makris ◽  
Kenneth J. Ryan
Keyword(s):  
Cyclic Amp ◽  
Granulosa Cells ◽  
Dose Response ◽  
Cyclic Gmp ◽  
Response Relationship ◽  
Dose Response Relationship ◽  
Cyclic Amp Accumulation ◽  
Relationship Of ◽  
Higher Doses ◽  
Cyclic Amp Synthesis

ABSTRACT Cyclic AMP and cyclic GMP accumulation in hamster granulosa cells as a function of gonadotrophin dose (LH or FSH) and time (0–30 min) was determined. The pattern of acute cyclic AMP accumulation was similar in LH and FSH stimulated granulosa cells, except that the cells were more sensitive to FSH than LH. There was a positive dose response relationship of cyclic AMP accumulation in LH and FSH stimulated cells. LH appeared to partially inhibit FSH stimulated cyclic AMP synthesis. Cyclic GMP accumulation was distinctly different in LH and FSH stimulated cells. An inverse dose response relationship of cyclic GMP to dose LH was observed, with only the lowest dose of LH (0.005 IU/ml) stimulating cyclic GMP synthesis. FSH at 0.005 IU/ml did not stimulate cyclic GMP synthesis, but at higher doses generated cyclic GMP in a positive dose-related manner. The results suggest that specificity of hormone action in granulosa cells may be governed in part by differential on cyclic AMP and cyclic GMP in these cells.

Changes in responsiveness of rat granulosa cells to prostaglandin E2 and follicle-stimulating hormone during culture

1983 ◽  
Vol 61 (6) ◽  
pp. 608-613 ◽  
Author(s):  
A. K. Goff ◽  
D. T. Armstrong
Keyword(s):  
Cyclic Amp ◽  
Granulosa Cells ◽  
Follicle Stimulating Hormone ◽  
Prostaglandin E ◽  
Dependent Manner ◽  
Isolated Cells ◽  
Progesterone Production ◽  
Dose Dependent ◽  
Stimulating Factor ◽  
Stimulation Of

The changes in responsiveness of granulosa cells to either FSH or prostaglandin E2 (PGE2), during culture of the cells, have been examined. In freshly isolated cells, FSH and PGE2 stimulated both cyclic AMP and progesterone production in a dose–dependent manner. In these cells, FSH stimulated cyclic AMP production to a greater extent than did PGE2. After the cells had been cultured for 2 days, neither FSH nor PGE2 stimulated progesterone production to any detectable extent. In these cells the ability of FSH to stimulate cyclic AMP was decreased, and that of PGE2 was increased markedly, such that PGE2 was far more effective than FSH in stimulating cyclic AMP. After culture of the cells for a further 2 days (4 days total), the FSH stimulation of cyclic AMP returned to that seen in freshly isolated cells, whereas the stimulation by PGE2 remained elevated. The acute stimulation of progesterone production could be restored by chronic exposure of the cells to either FSH or PGE2. These results demonstrate that dramatic changes in responsiveness of granulosa cells take place during culture. The results also suggest that some stimulating factor must be present to maintain the steroidogenic capabilities of the cells. Without this, although the cells are able to produce cyclic AMP in response to FSH, they cannot produce progesterone.

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