ABSTRACT
The activity of adenylate deaminase, adenylate phosphatase and adenosine deaminase, as well as the endogenous content of adenine nucleotides, was examined in the heart of ageing chickens. In new-born (1-day-old) and young (20-day-old) chickens, AMP degradation in the heart seems to proceed preferentially through deamination, while in adult (1-year-old) through dephosphorylation. Compared with the adult heart, a 2-year-old one exhibits a decline of AMP catabolism. The total adenine nucleotide content and the concentration of ATP are higher in adult and aged chicken hearts, than in new-born and young ones. Adaptive mechanisms might occur in the heart of ageing chickens to ensure an adequate availability of adenine nucleotides.
Subject(s)
Adenosine Monophosphate/metabolism , Aging/metabolism , Chickens/metabolism , Myocardium/metabolism , 5'-Nucleotidase/metabolism , Adenosine Deaminase/metabolism , Adenosine Diphosphate/metabolism , Adenosine Triphosphate/metabolism , Animals , Animals, Newborn , Magnesium Chloride/pharmacologyABSTRACT
The influence of [Mg2+] on the basal or stimulated activity of adenylate cyclase from the hearts of young (1 month old) and aged (24 months old) rats has been investigated in vitro. The basal activity of cardiac adenylate cyclase, and its responsiveness to stimulatory or inhibitory effectors, declined with age. This is probably due to alterations at the catalytic moiety of the signal transduction system, such as an impairment in the affinity of the catalytic moiety for ATP and a lower capacity of the catalytic moiety to bind activated stimulatory (Gs) or inhibitory (Gi) guanine nucleotide binding proteins. Compared to the enzyme from the heart of aged rats, unstimulated adenylate cyclase from the heart of young rats was more sensitive to an increase in [Mg2+] in the incubation mixture, as shown by a greater increase in basal activity and in the affinity of the enzyme for ATP. An increase in [Mg2+] counteracted the inhibitory effect of spermine on adenylate cyclase more effectively in young rats than in aged rats. On the other hand, an increase in [Mg2+] facilitated the stimulation of adenylate cyclase by Gpp(NH)p, isoproterenol and forskolin more in aged rats than in young rats. GDP beta S prevented the positive effect of high [Mg2+] on the stimulation of adenylate cyclase by forskolin, suggesting that an increased [Mg2+] favors the activation of Gs or the formation of functional complexes between the catalytic moiety and Gs. We suggest that aging leads to a higher requirement for Mg2+ at the allosteric site on the catalytic moiety whose occupancy is essential for the full expression of stimulated activity.
Subject(s)
Adenylyl Cyclases/metabolism , Aging/metabolism , Magnesium/physiology , Myocardium/enzymology , Adenosine Triphosphate/metabolism , Adenylyl Cyclase Inhibitors , Animals , GTP-Binding Proteins/physiology , In Vitro Techniques , Male , Rats , Rats, Wistar , Signal Transduction/physiologyABSTRACT
The metabolic and functional activity of the heart closely depends on cAMP and therefore on the integrity of adenylate cyclase (AC) system. Alterations of this signal transduction system might be co-responsible for the impairment of cardiac performance observed with aging. Evidence is here provided that basal activity of cardiac membrane-bound (48,000 x g) AC significantly declines with the age of the rat (1, 12, 24 month-old). This is accompanied with the decrease of cAMP content, which leads to the fall of cAMP/cGMP molar ratio a possible final determinant of cardiac performance. Kinetic analyses indicate that aging is associated with a net increase of the Km of a cardiac AC, while the Vmax is unaffected. Besides, the response in vitro of AC from 24-month-old heart to the inhibitor spermine or a different stimulants, such as Gpp (NH) p, isoproterenol, PGE1 or forskolin, is significantly lower than that of AC from 1 month-old one. The suggestion is made that aging causes an impairment in the capability of the catalytic moiety of cardiac AC to make functional complexes with activated guanine nucleotide binding proteins.
Subject(s)
Adenylyl Cyclases/metabolism , Aging/metabolism , Myocardium/enzymology , Adenylyl Cyclases/analysis , Adenylyl Cyclases/drug effects , Aging/drug effects , Animals , Cyclic AMP/analysis , Cyclic GMP/analysis , Male , Myocardium/chemistry , Rats , Rats, WistarABSTRACT
The exposure of quiescent cultures of cardiac cells to 1 microM spermine for 2 hours leads to an increase of the content of intracellular polyamines and to a 40% decrease of basal adenylate cyclase activity. The response of adenylate cyclase to stimulation by PGE1 is reduced by about 50% after spermine treatment. The effects of the amine on adenylate cyclase are completely prevented by pretreating the cells with pertussis toxin which blocks the activation of the inhibitory guanine binding protein (Gi). In vitro experiments with adenylate cyclase from cells pre-treated with pertussis toxin show that spermine fails to reduce basal enzyme activity and to counteract the stimulation by PGE1 or forskolin. Cholera toxin, which blocks the deactivation of the stimulatory protein (Gs), does not influence the effects of spermine either in vivo or in vitro. The results suggest that spermine acts through the activation of Gi. This hypothesis is supported by the fact that, in vitro, the inhibition of stimulated adenylate cyclase by the amine is synergistic with that of a stable analog of GDP, GDP beta S, which causes deactivation of Gs.
