title: Cytochemical studies on sarcolemma: Na+, K+-adenosine triphosphatase and adenylate cyclase.
abstract: Two highly lead-sensitive ATPases, Na+,K+-ATPase and adenylate cyclase, can be demonstrated cytochemically by the lead precipitation technique in briefly prefixed tissue, provided that the free Pb2+ concentration in the incubation medium is kept below 0.1 mM by a heavy metal chelator. Under conditions suitable for Na+,K+-ATPase activity precipitation of final reaction product (lead phosphate) at the sarcolemma of cardiac muscle is abolished by 0.1-1mM ouabain. In contrast, reaction product deposition at the intramuscular part of the plasma membrane and at intracellular sites is not noticeably affected by the glycoside. These findings indicate either that the sarcolemma is the exclusive location of Na+,K+-ATPase in cardiac muscle or that the presence of the enzyme at other loci is masked by active Na+,K+-independent, ouabain resistant ATPases. Under conditions favoring adenylate cyclase activity, precipitation by Pb2+ of orthophosphate derived, with the help of added cyclic nucleotide phosphodiesterase and 5'-nucleotidase, from cyclic AMP formed from adenylyl imidodiphosphate (AMP-PNP) is seen after prolonged incubation in myocardial cells along the entire course of the plasma membrane and also at the transverse tubules and is particularly intense at the tight junction regions of the intercalated disks. Ouabain has no effect on these reactions. Reaction product deposition is also observed at the sarcolemma in red skeletal muscle and at the terminal cisternae of the sarcoplasmic reticulum in white skeletal muscle, where the reaction is intensified by adrenaline. Sarcoplasmic reticulum of cardiac and of red skeletal muscle exhibits only relatively weak staining attributable to cyclic AMP formation. These observations are in agreement with the results of tissue fractionation studies according to which the plasma membrane is the chief site of adenylate cyclase in heart and in red, but not white skeletal muscle.
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D002645:Chickens; D006352:Heart Ventricles / Q000201:enzymology / Q000648:ultrastructure; D007854:Lead; D008854:Microscopy, Electron; D009206:Myocardium / Q000201:enzymology*; D010042:Ouabain / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D012508:Sarcolemma / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D012964:Sodium / Q000494:pharmacology
chemical_list: D007854:Lead; D010042:Ouabain; D012964:Sodium; D000251:Adenosine Triphosphatases; D000262:Adenylyl Cyclases; D011188:Potassium