cAMP signalling mechanisms with aging in the Ceratitis capitata brain.

Aging has been associated with alterations in protein phosphorylation. This study was undertaken to examine eventual changes in cAMP-dependent protein kinase (PKA) activity and enzyme regulatory subunit levels from the dipterous Ceratitis capitata brain with postmaturational aging and senescence. PKA activity was determined in cytosolic and membrane fractions of the C. capitata brain during the adult stage of the insect lifespan. PKA activity markedly increased at the first stages of the life of the fly both in cytosol and in membranes. A lower peak of PKA activity was evident both in particulate and cytosolic fractions in the terminal phase of the life of the fly. Thus, PKA activity was significantly higher in the brain of mature flies when compared to the brain of aged flies. It is possible that increases in cAMP-dependent protein phosphorylation levels characterize the terminal aging process in the insect nervous tissue. On the other hand, levels of regulatory (R) subunit were also measured in membranes and cytosol by immunoblotting. Cytosolic regulatory subunit levels were more elevated near the terminal phase of life, whereas in membranes, regulatory subunit levels decrease in senescence in parallel with particulate PKA activity. The increased R subunit level in cytosol may reflect a cellular response mechanisms to down-regulate the kinase system in aged flies.

Levels and activity of brain protein kinase C alpha and zeta during the aging of the medfly.

Brain protein kinase C (PKC) activity, as well as PKC alpha and PKC zeta levels detected by immunoblotting, were monitored during the lifespan of the Mediterranean fruit-fly Ceratitis capitata. PKC activity increased in the particulate fraction during the last stages of the life of C. capitata. Immunoblotting studies with an anti-PKC alpha antibody also demonstrated increased enzyme levels in the particulate fraction. Cytosolic levels of PKC zeta decreased in the terminal phase of the lifespan of the fly, whereas levels of membrane-bound PKC zeta increased at that stage. Results thus indicate that during C. capitata final phase of life a translocation of PKC alpha and PKC zeta to the particulate fraction occurs, and therefore both kinases could be involved in the terminal process of this fruit-fly.

Adenylyl cyclase system is affected differently by endurance physical training in heart and adipose tissue.

Adaptive changes in the beta-adrenergic adenylyl cyclase (AC) system in response to endurance training were studied in heart and adipose tissue. Training was performed by making male Wistar rats run on a motor-driven treadmill. The changes following exercise training were opposite in the two tissue studied. The density of beta-adrenergic receptors in left ventricular membranes of trained rats showed a marked decrease. Comparison of AC activities in cardiac membranes prepared from trained and sedentary rats revealed a depressing effect of endurance training on: 1. the beta-adrenergic stimulatory pathway and the inhibition of AC via receptor; 2. the Gs component and the Gs-adenylyl cyclase coupling, as shown by the response of adenylyl cyclase to GppNHp and NaF; and 3. the enzyme catalytic activity in the presence of Mn2+ or forskolin. The levels of Gsalpha subunits in the left ventricle, as measured in terms of ADP-ribosylated and immunologically reactive proteins, were decreased by endurance exercise, whereas immunodetectable levels of Gialpha2 increased in the membranes of trained myocardium. In contrast to the diminished sensitivity that characterizes the behavior of the cardiac beta-adrenergic-AC system, endurance physical training increased sensitivity of this signal transduction system in adipose tissue. Thus, the density of beta-ARs as well as AC activity and the beta-adrenergic stimulatory pathway were increased in adipose membranes of trained rats compared with the corresponding sedentary controls. In addition, the levels of Gsalpha subunits were higher in the adipose plasma membranes of trained rats. However, immunodetectable levels of Gi1alpha and Gi3alpha increased with training, whereas the amount of Gi2alpha decreased in membranes of trained rats. In conclusion, the present study shows that chronic exercise is associated with a tissue-specific adaptation of the beta-adrenergic AC system.

Phosphatidylcholine-phospholipase C mediates the induction of nerve growth factor in cultured glial cells.

Addition of phosphatidylcholine-hydrolyzing phospholipase C (PC-PLC) to cultured glial cells increased the levels of nerve growth factor (NGF) mRNA and the amount of cell-secreted NGF. The effect of PC-PLC was 2.5 times higher than that elicited by 4 beta-phorbol 12 beta-myristate 13 alpha-acetate. In cells in which protein kinase C (PKC) was fully inhibited or downregulated, the effect of PC-PLC was reduced-though still evident-and similar to that exerted by sphingosine. Results thus indicate that PC-PLC induces the synthesis of NGF by glial cells by a PKC-dependent and PKC-independent mechanisms.