scholarly journals A study on sensing and adaptation in Dictyostelium discoideum: guanosine 3',5'-phosphate accumulation and light-scattering responses.

1983 ◽  
Vol 96 (6) ◽  
pp. 1566-1570 ◽  
Author(s):  
B Wurster ◽  
U Butz

Cells of Dictyostelium discoideum respond to extracellular cyclic AMP with marked changes in intracellular cyclic GMP levels and light scattering. In this work, defined temporal increases in cyclic AMP were produced by the continuous addition of cyclic AMP to agitated suspensions of cells; concomitant hydrolysis of cyclic AMP by the cells subsequently established a constant, steady state concentration. The cells responded to the initial increase in extracellular cyclic AMP with a rapid increase in the intracellular cyclic GMP concentration and a rapid decrease in light scattering. At cyclic AMP input rates of 0.5-5 nM X s-1, the fast reactions of cyclic GMP and light scattering had already relaxed while the cyclic AMP concentration in the cell suspension was still increasing. The cells responded to constant concentrations of cyclic AMP with constant elevated cyclic GMP concentrations and constant decreased levels of light scattering. Our results are consistent with the existence of two types of perception systems, one of which adapts to constant stimuli and one of which does not adapt.

1987 ◽  
Vol 87 (5) ◽  
pp. 723-730
Author(s):  
B. Wurster ◽  
R. Mohn

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.


FEBS Letters ◽  
1977 ◽  
Vol 79 (2) ◽  
pp. 331-336 ◽  
Author(s):  
José M. Mato ◽  
Peter J.M. Van Haastert ◽  
Frans A. Krens ◽  
Els H. Rhunsburger ◽  
Fred C.P.M. Dobbe ◽  
...  

1993 ◽  
Vol 10 (6) ◽  
pp. 991-996 ◽  
Author(s):  
Ari Sitaramayya ◽  
Lorraine Lombardi ◽  
Alexander Margulis

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.


1974 ◽  
Vol 18 (6) ◽  
pp. 509-515 ◽  
Author(s):  
G. Chader ◽  
R. Fletcher ◽  
M. Johnson ◽  
R. Bensinger

1982 ◽  
Vol 152 (1) ◽  
pp. 232-238
Author(s):  
P J van Haastert ◽  
F J Pasveer ◽  
R C van der Meer ◽  
P R van der Heijden ◽  
H van Walsum ◽  
...  

Chemotactic stimulation of vegetative or aggregative Dictyostelium discoideum cells induced a transient elevation of cyclic GMP levels. The addition of chemoattractants to postvegetative cells by pulsing induced phosphodiesterase activity. The following lines of evidence suggest a messenger function for cyclic GMP in the induction of phosphodiesterase: (i) Folic acid and cyclic AMP increased cyclic GMP levels and induced phosphodiesterase activity. (ii) Cyclic AMP induced both cyclic GMP accumulation and phosphodiesterase activity by binding to a rate receptor. (iii) The effects of chemical modification of cyclic AMP or folic acid on cyclic GMP accumulation and phosphodiesterase induction were closely correlated. (iv) A close correlation existed between the increase of cyclic GMP levels and the amount of phosphodiesterase induced, independent of the type of chemoattractant by which this cyclic GMP accumulation was produced. (v) Computer simulation of cyclic GMP binding to intracellular cyclic GMP-binding proteins indicates that half-maximal occupation by cyclic GMP required the same chemoattractant concentration as did half-maximal phosphodiesterase induction.


1987 ◽  
Vol 88 (3) ◽  
pp. 379-388
Author(s):  
M.B. Coukell ◽  
A.M. Cameron

Previous studies have suggested that cyclic GMP and/or Ca2+ might function as second messengers in the induction by exogenous cyclic AMP of the cyclic AMP phosphodiesterase (PD) in Dictyostelium discoideum. To assess further the role of Ca2+ in PD induction we examined the effect on this process of a number of putative Ca2+-channel blockers. At relatively low micromolar concentrations, TMB-8, nicardipine, nifedipine, diltiazem and verapamil all altered cell shape and inhibited PD induction in a similar dose-dependent fashion. Concentrations of these drugs that abolished PD induction had no effect on cell viability; however, higher concentrations reduced viability and caused cell lysis. All effects of these compounds on the cells were antagonized at least partially by 5–10 mM-Ca2+. Other cations tested were considerably less effective. Like the organic inhibitors, La3+ also altered cell shape, inhibited PD induction and reduced cell viability at elevated concentrations, but its effect on the cells appeared to be more complex. Inhibition of PD induction by the organic antagonists could not be attributed solely to an impaired uptake of extracellular Ca2+, a reduction of ATP pools in the cells or a direct effect on calmodulin. Concentrations of TMB-8 that inhibited PD induction had little effect on the cyclic GMP response. Therefore, this compound did not inhibit PD induction indirectly by blocking cyclic GMP synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)


1986 ◽  
Vol 234 (2) ◽  
pp. 325-334 ◽  
Author(s):  
N J Pyne ◽  
M E Cooper ◽  
M D Houslay

Two enzymes displaying cyclic GMP-stimulated cyclic AMP phosphodiesterase activity were purified from rat liver to apparent homogeneity: a ‘particulate enzyme’ found as an integral membrane protein associated with the plasma membrane, and a ‘soluble’ enzyme found in the cytosol. The physical properties of these enzymes were very similar, being dimers of Mr 134,000, composed in each instance of two subunits of Mr = 66,000-67,000. Both enzymes showed similar kinetics for cyclic AMP hydrolysis. They are both high-affinity enzymes, with kinetic constants for the particulate enzyme of Km = 34 microM and Vmax. = 4.0 units/mg of protein and for the cytosolic enzyme Km = 40 microM and Vmax. = 4.8 units/mg of protein. In both instances hydrolysis of cyclic AMP appeared to show apparent positive co-operativity, with Hill coefficients (happ.) of 1.5 and 1.6 for the particulate and cytosolic enzymes respectively. However, in the presence of 2 microM-cyclic GMP, the hydrolysis of cyclic AMP obeyed Michaelis kinetics (happ. = 1) for both enzymes. The addition of micromolar concentrations of cyclic GMP had little effect on the Vmax. for cyclic AMP hydrolysis, but lowered the Km for cyclic AMP hydrolysis to around 20 microM in both cases. However, at low cyclic AMP substrate concentrations, cyclic GMP was a more potent activator of the particulate enzyme than was the soluble enzyme. The activity of these enzymes could be selectively inhibited by cis-16-palmitoleic acid and by arachidonic acid. In each instance, however, the hydrolysis of cyclic AMP became markedly more sensitive to such inhibition when low concentrations of cyclic GMP were present. Tryptic peptide maps of iodinated preparations of these two purified enzyme species showed that there was considerable homology between these two enzyme forms.


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