Renaissance of cytochemical localization of membrane ATPases in the myocardium.
ATPases of cardiac cells are known to be among the most important enzymes to maintain the fluxes of vital cations by hydrolysis of the terminal high-energy phosphate of ATP. Biochemically the activities of Ca(2+)-pump ATPase, Ca2+/Mg(2+)-ecto ATPase, Na+,K(+)-ATPase and Mg(2+)-ATPase are determined in homogenates and isolated membranes as well as in myofibrillar and mitochondrial fractions of various purities. Such techniques permit estimation of enzyme activities in vitro under optimal conditions without precise enzyme topography. On the other hand, cytochemical methods demonstrate enzyme activity in situ, but not under optimal conditions. Until recently several cytochemical methods have been employed for each enzyme in order to protect its specific activity and precise localization but the results are difficult to interpret. To obtain more consistent data from biochemical and cytochemical point of view, we modified cytochemical methods in which unified conditions for each ATPase were used. The fixative solution (1% paraformaldehyde-0.2% glutaraldehyde in 0.1 M Tris Base buffer, pH 7.4), the same cationic concentrations of basic components in the incubation medium (0.1 M Tris Base, 2 mM Pb(NO2)3, 5 mM MgSO4, 5 mM ATP) and selective stimulators or inhibitors were employed. The results reveal improved localization of Ca(2+)-pump ATPase, Na(+)-K+ ATPase and Ca2+/Mg(2+)-ecto ATPase in the cardiac membrane.
D000251:Adenosine Triphosphatases / Q000032:analysis* / Q000037:antagonists & inhibitors; D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase / Q000032:analysis / Q000037:antagonists & inhibitors; D000252:Calcium-Transporting ATPases / Q000032:analysis / Q000037:antagonists & inhibitors; D002462:Cell Membrane / Q000201:enzymology; D004533:Egtazic Acid / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008854:Microscopy, Electron; D009206:Myocardium / Q000201:enzymology* / Q000648:ultrastructure; D015388:Organelles / Q000201:enzymology; D051381:Rats; D017208:Rats, Wistar; D000254:Sodium-Potassium-Exchanging ATPase / Q000032:analysis; D013729:Terpenes / Q000494:pharmacology; D019284:Thapsigargin
D004791:Enzyme Inhibitors; D013729:Terpenes; D004533:Egtazic Acid; D019284:Thapsigargin; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases; D000254:Sodium-Potassium-Exchanging ATPase