scholarly journals Dibutyryladenosine 3′:5′-cyclic monophosphate-mediated changes in rat cells involve macromolecular alterations in vinblastine-precipitable proteins

1978 ◽  
Vol 174 (3) ◽  
pp. 1071-1074
Author(s):  
A T Meza ◽  
M Rieber
Keyword(s):  
Molecular Weight ◽  
Cyclic Amp ◽  
Cell Cultures ◽  
Cyclic Nucleotide ◽  
Dibutyryl Cyclic Amp ◽  
Cyclic Monophosphate ◽  
Three Components

ts-NT3-KR rat cell cultures show the loss of three components in the molecular-weight region 200,000–250,000 when exposed to dibutyryl cyclic AMP, under conditions of both restriction and expression of the transformed phenotype. Vinblastine is able to precipitate preferentially from control cultures the species that are decreased by exposure to the cyclic nucleotide. Serum-starved cultures exposed to dibutyryl cyclic AMP reveal differences in their vinblastine precipitates, depending on whether the expression of the transformation phenotype is restricted or not.

scholarly journals Influence of glucagon, 6-N,2′-O-dibutyryladenosine 3′:5′-cyclic monophosphate and triamcinolone on the arginine synthetase system in perinatal rat liver

1972 ◽  
Vol 126 (1) ◽  
pp. 89-98 ◽  
Author(s):  
A L. Schwartz
Keyword(s):  
Cyclic Amp ◽  
Rat Liver ◽  
Cyclic Nucleotide ◽  
Time Course ◽  
Fold Increase ◽  
Dibutyryl Cyclic Amp ◽  
System Activity ◽  
Cyclic Monophosphate ◽  
Foetal Rat ◽  
Postnatal Rats

1. The administration of triamcinolone (19–190μg/animal) to postnatal rats increased the arginine synthetase system activity 1.2–2.5-fold above control values 24h after exposure to the hormone. Cortisol (hydrocortisone), however, increased the arginine synthetase system activity only when larger (190μg/animal) or repeated daily doses were given. Glucagon (100μg/animal) stimulated arginine synthetase system activity only after the second postnatal day. None of these agents increased the activity in 19.5–21.5-day foetuses after intrauterine administration. 2. The viability of foetal rat liver explants maintained in organ culture for up to 54h was validated both by ultramicroscopic examination and by incorporation of radioactive leucine and orotic acid. 3. In organ cultures of foetal rat liver explants (18.5 days to term), triamcinolone (20μg/ml of medium) evoked a 2.8–4.3-fold increase after 24h of incubation. This increase was completely inhibited by actinomycin D (25μg/ml) or cycloheximide (10μg/ml). Cortisol (5–50μg/ml) or glucagon (0.067–67μg/ml) also increased the arginine synthetase system activity above the respective control values, but there was no increase in activity with insulin (0.05–0.25i.u./ml). 4. Maximum concentrations of glucagon (67μg/ml), dibutyryl cyclic AMP (6-N,2′-O-dibutyryladenosine 3′:5′-cyclic monophosphate) (0.1mm) and triamcinolone (20μg/ml) incubated for 24h with foetal rat liver explants each produced between a two-and three-fold increase in the activity of the arginine synthetase system. Combinations of maximum amounts of glucagon and the cyclic nucleotide did not produce a greater effect than either agent alone. However, the combination of dibutyryl cyclic AMP with triamcinolone appeared to produce somewhat less than additive effects. 5. The effects of the cyclic nucleotide and triamcinolone were evident after 12h of incubation and increased steadily throughout the 24h of observation. This time-course of increased enzyme activity is very much slower than that reported for the induction of other enzymes in explant cultures of foetal rat liver.

Cyclic Nucleotide Levels during Spontaneous Uterine Contractions

1974 ◽  
Vol 52 (3) ◽  
pp. 763-767 ◽  
Author(s):  
Jack Diamond ◽  
Diane K. Hartle
Keyword(s):  
Cyclic Amp ◽  
Cyclic Gmp ◽  
Cyclic Nucleotide ◽  
Spontaneous Contraction ◽  
Rat Uterus ◽  
Cyclic Monophosphate ◽  
Tissue Levels ◽  
Uterine Contractions ◽  
Spontaneous Contractions ◽  
Isolated Rat

Tissue levels of adenosine 3′,5′-cyclic monophosphate (cyclic AMP) and guanosine 3′,5′-cyclic monophosphate (cyclic GMP) were determined in uterine muscles frozen at various points during spontaneous contraction–relaxation cycles. No significant changes in cyclic nucleotide levels were detected at any of the stages of contraction studied. Exposure of uterine segments to 1 μM isoproterenol resulted in an eightfold increase in cyclic AMP levels but no change in cyclic GMP, whereas exposure to 2 mM theophylline resulted in a doubling of cyclic GMP levels and a 42% increase in cyclic AMP content. Thus, the methods used were capable of detecting changes in cyclic nucleotide levels when they did occur. It was concluded that changes in cyclic nucleotide levels do not play a role in the initiation or regulation of spontaneous contractions of isolated rat uterus.

scholarly journals The regulation of growth hormone secretion from the isolated rat anterior pituitary in vitro. The role of adenosine 3′:5′-cyclic monophosphate

1971 ◽  
Vol 124 (4) ◽  
pp. 815-826 ◽  
Author(s):  
R. B. Lockhart Ewart ◽  
K. W. Taylor
Keyword(s):  
Growth Hormone ◽  
Cyclic Amp ◽  
Early Phase ◽  
Late Phase ◽  
Hormone Secretion ◽  
Growth Hormone Secretion ◽  
Hormone Release ◽  
Dibutyryl Cyclic Amp ◽  
Cyclic Monophosphate

1. The release of growth hormone from isolated fragments of rat anterior pituitary tissue incubated in vitro was studied by employing a double-antibody radioimmunoassay. 2. In the absence of added stimuli, two phases of hormone release could be distinguished, an early phase of 2h duration and a subsequent late phase. In the early phase, hormone release was rapid but could be significantly decreased by calcium depletion and by 2,4-dinitrophenol whereas the rate of release in the late phase was uninfluenced by these incubation conditions. These results have been interpreted as indicating the existence of a secretory component in the early phase of release. 3. In subsequent experiments, the effects of various agents on the rate of hormone output during the late phase of incubation were investigated. Hormone release was increased by theophylline and by dibutyryl cyclic AMP (N6-2′-O-dibutyryl-adenosine 3′:5′-cyclic monophosphate), the response to both of these agents being related to the concentration of the stimulant employed. 4. The stimulation of growth hormone output by theophylline was significantly decreased by calcium deprivation and by 2,4-dinitrophenol. The response to dibutyryl cyclic AMP was diminished by 2,4-dinitrophenol, iodoacetate and 2-deoxyglucose but not by malonate or colchicine. 5. Arginine, β-hydroxybutyrate, albumin-bound palmitate and variation in the glucose concentration of the incubation medium over a wide range were without any statistically significant effect on the rate of hormone release from either control pituitary fragments or those subject to secretory stimulation by dibutyryl cyclic AMP. 6. It is suggested that the regulation of growth hormone secretion is mediated by cyclic AMP (adenosine 3′:5′-cyclic monophosphate). The secretion observed in response to cyclic AMP requires the presence of ionized calcium and a source of metabolic energy but is independent of pituitary protein synthesis de novo. The integrity of the glycolytic pathway of glucose metabolism appears to be essential for cyclic AMP-stimulated growth hormone secretion to occur.

scholarly journals Stimulation by 6-N,2′-O-dibutyryladenosine 3′:5′-cyclic monophosphate and glucocorticoids of phosphoenolpyruvate carboxykinase in the isolated perfused rat liver (Short Communication)

1974 ◽  
Vol 142 (3) ◽  
pp. 691-693 ◽  
Author(s):  
Wieland B. Huttner ◽  
Wilhelm Krone ◽  
Hans J. Seitz ◽  
Wolfgang Tarnowski
Keyword(s):  
Cyclic Amp ◽  
Cyclic Nucleotide ◽  
Time Course ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Direct Action ◽  
Isolated Perfused Rat Liver ◽  
Dibutyryl Cyclic Amp ◽  
Low Protein ◽  
Additive Increase

Dibutyryl cyclic AMP stimulated the activity of phosphoenolpyruvate carboxykinase in perfused livers of rats, fed on a low-protein diet, linearly over a 6h period. The enzyme activity was also significantly elevated by dexamethasone, the effect being considerably lower than that of the cyclic nucleotide. Since the time-course of phosphoenolpyruvate carboxykinase activity in response to dibutyryl cyclic AMP resembled that observed after dibutyryl cyclic AMP injection into intact animals, it is suggested that induction of the enzyme in vivo is due to a direct action of the cyclic nucleotide on the liver. Combined administration of dibutyryl cyclic AMP and glucocorticoids did not lead to an additive increase of liver phosphoenolpyruvate carboxykinase activity, either in vivo or in the perfused organ.

scholarly journals Phosphoproteins associated with cyclic nucleotide stimulation of ciliary motility in Paramecium

1990 ◽  
Vol 95 (2) ◽  
pp. 219-230
Author(s):  
N.M. Bonini ◽  
D.L. Nelson
Keyword(s):  
Molecular Weight ◽  
Protein Kinase ◽  
Cyclic Amp ◽  
Cyclic Gmp ◽  
Cyclic Nucleotide ◽  
Protein Kinase Activity ◽  
Dependent Protein Kinase ◽  
Permeabilized Cells ◽  
Ciliary Motility ◽  
Dependent Protein

Permeabilized, MgATP-reactivated cells of Paramecium (models) respond to cyclic AMP and cyclic GMP by increasing forward swimming speed. In association with the motile response, cyclic AMP and 8-bromo-cyclic GMP (8-Br-cyclic GMP) stimulated protein phosphorylation. Cyclic AMP addition to permeabilized cells reproducibly stimulated the phosphorylation of 10 proteins, ranging in molecular weight from 15 to 110K (K = 10(3) Mr). 8-Br-cyclic GMP, which selectively activates the cyclic GMP-dependent protein kinase of Paramecium, stimulated the phosphorylation of a subset of the proteins phosphorylated by cyclic AMP. Ca2+ addition caused backward swimming and stimulated the phosphorylation of four substrates, including a 25K target that may also be phosphorylated in response to cyclic nucleotide addition. Ba2+ and Sr2+ also induced backward swimming, but did not cause detectable phosphorylation. To identify ciliary targets of cyclic nucleotide-dependent protein kinase activity, permeabilized cells were deciliated following reactivation of motility with Mg-[gamma-32P]ATP in the presence or absence of cyclic nucleotide. Soluble proteins of the deciliation supernatant were enriched in 15 cyclic AMP-stimulated phosphoproteins, ranging in molecular weight from 15 to 95K. Most of the ciliary substrates were axonemal and could be released by high salt solution. A 29K protein that copurified in sucrose gradients with the 22S dynein, and a high molecular weight protein (greater than 300K) in the 19 S region were phosphorylated when cyclic AMP was added to permeabilized, motile cells. These data suggest that regulation of ciliary motility by cyclic AMP may include phosphorylation of dynein-associated proteins.

scholarly journals REGULATION OF PHOSPHOENOLPYRUVATE CARBOXYKINASE AND TYROSINE TRANSAMINASE IN HEPATOMA CELL CULTURES

1974 ◽  
Vol 60 (1) ◽  
pp. 181-191 ◽  
Author(s):  
Han van Rijn ◽  
Marinus M. Bevers ◽  
Roeland van Wijk ◽  
Wesley D. Wicks
Keyword(s):  
Growth Rate ◽  
Cyclic Amp ◽  
Dna Synthesis ◽  
Cell Cultures ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Hepatoma Cell ◽  
Transaminase Activity ◽  
Dibutyryl Cyclic Amp ◽  
Tyrosine Transaminase ◽  
Htc Cells

The ability of N6, O2'-dibutyryl cyclic AMP (DBcAMP) to regulate a number of metabolic events in four lines of cultured rat hepatomas has been examined. Although dexamethasone induces tyrosine transaminase in all four lines, DBcAMP induces this enzyme normally only in H35 cells. A slight increase in transaminase activity was seen with MH1C1 cells and HTC cells, but no effect was detectable in RLC cells. In contrast, phosphoenolpyruvate carboxykinase activity is increased by both agents in H35 and MH1C1 cells, but neither had any effect in HTC or RLC cells. DBcAMP caused a rapid inhibition of the growth rate and DNA synthesis and an increase in protein content in both H35 and MH1C1 cells but not in HTC or RLC cells. The effect of DBcAMP on DNA synthesis in MH1C1 cells could be reversed by deoxycytidine as is also the case with H35 cells. The resistance of HTC and RLC cells to DBcAMP was not due to reduced uptake or deacylation as judged by studies with [3H]DBcAMP. The cyclic nucleotide appears to enter the cells by passive diffusion as the intracellular concentration approaches that in the medium within 30–60 min. Possible explanations for the differential responses observed are discussed.

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