Ca2+ extrusion in aged smooth muscle cells.
We investigated the effects of aging in Ca(2+) extrusion mechanisms in smooth muscle bladder cells from 4 and 20-24-month-old guinea pigs using fluorescence microscopy and fura-2. Cells were challenged with a pulse of KCl immediately before perfusion with a Ca(2+) free solution containing no inhibitors (control, untreated cells) or inhibitors of plasma membrane Ca(2+) pump (PMCA, 1mM La(3+)), Na(+)/Ca(2+) exchanger (NCX, 1 microM SEA0400) or the sarcoendoplasmic Ca(2+) pump (SERCA, 1 microM thapsigargin). Treatment of young adult cells with the inhibitors allowed estimating a relative contribution of 55% for NCX, 27% for PMCA and 31% for SERCA. Combination of two inhibitors at the same time showed the presence of interaction between extrusion mechanisms. In aged cells the [Ca(2+)](i) extrusion was impaired due to decrease of PMCA activity, as revealed by the loss of effect of La(3+), and to inhibitory interactions between NCX and SERCA activities, indicated by acceleration of decay in response to their respective inhibitors. In conclusion, in smooth muscle cells aging decreases the overall Ca(2+) extrusion activity and modifies the interactions between the activities of the main Ca(2+) removing mechanisms.
Gomez-Pinilla
Pedro J
PJ
Department of Physiology, University of Extremadura, Campus Universitario, Fac Veterinary, 10071 Caceres, Spain.
Pozo
Maria J
MJ
Baba
Akemishi
A
Matsuda
Toshio
T
Camello
Pedro J
PJ
D000818:Animals; D001692:Biological Transport; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels / Q000378:metabolism; D016922:Cellular Senescence; D016257:Fura-2; D006168:Guinea Pigs; D008856:Microscopy, Fluorescence; D032389:Myocytes, Smooth Muscle / Q000378:metabolism / Q000502:physiology*; D001743:Urinary Bladder / Q000166:cytology*
D002121:Calcium Channel Blockers; D015220:Calcium Channels; D002118:Calcium; D016257:Fura-2