A B S T R A C T The possibility has been explored that decreases of adenylate cyclase may explain diminished hormone sensitivity of adipose tissue with aging. Isolated cells were prepared from epididymal fat pads of rats 1-, 2-, 6-, 12-, and 24-mo-old, fixed in OsO, and sized and counted with a Coulter apparatus. Adenylate cyclase was assayed in cell membranes (ghosts) using [ra-P] 6 and 24 mo per cell. Glucagon stimulated adenylate cyclase 4.5-fold relative to basal in the 1-mo-old rat, but its effect then rapidly decreased and was absent by 12 mo. The fourfold stimulation by ACTH noted in the 1-mo-old animals decreased gradually with age but was still twice basal at 24 mo. Since no significant change of cell size occurred after 6 mo, diminished hormone sensitivity with senescence cannot be related to cell size. Similar age-related patterns of hormonal activation were evoked by 5'-guanylyl-imidodiphosphate [GMP-P(NH)P], a nucleotide analogue which increased both basal-and hormone-activated enzyme at all ages studied. Dose-response curves to hormones, fluoride, and GMP-P(NH)P were not affected by age. High Mg++ (50 mM) in the presence of GMP-P-(NH) P stimulated adenylate cyclase to levels greater than with fluoride, but a similar loss of activity with aging was still observed. Loss of hormone receptors may partially explain the age-related decreases of glucagon and ACTH-sensitive adenylate cyclase, but decreased basal-, epinephrine-, fluoride-, and GMP-P-(NH)P-stimulated responses suggest loss of the catalytic component of the adenylate cyclase enzyme complex in the aging fat cell membranes.
INTRODUCTIONIn the past, most studies of aging and hormones have dealt with effects of hormones on the processes of aging or, more often, with effects of aging on hormone secretion and metabolism (1). More recently, however, effects of aging on hormone responsiveness or sensitivity have come under investigation. A number of postmaturational changes of this type have now been identified. In view of the variety of mechanisms involved in the actions of hormones of different types, it is not surprising that a mixed picture has emerged (2). For the many hormones whose mechanism of action involves activation of adenylate cyclase, relatively little information on age relationships is as yet available. To extend our information in this area, and with the hope of being able eventually to relate the findings to physiologic events, we have undertaken studies of adenylate cyclases in a variety of animal and human tissues.