Influence of ionic composition, buffering agent, and pH on the histochemical demonstration of myofibrillar actomyosin ATPase.
The influence of the composition of the preincubation medium on the histochemical demonstration of myofibrillar actomyosin ATPase, including a variety of carboxylic acid and non-carboxylic acid buffering compounds and neutral salts, was studied. In inorganic salt-free systems the rate of the activation of type I fibers and inactivation of type II fibers was accelerated when the carboxylic acids had longer chain length or multiple carboxyl groups. Of these factors, the number of carboxyl groups was dominant with a 100 mM citrate buffer producing a sharp differentiation between fiber types. In contrast, the time course of the response was exceptionally long in an acetate buffer. The time course of the ATPase reaction was also modified by other buffers at pH 4.60. The most notable were an ascorbate - glycine buffer system which produced little or no deviation from the alkaline preincubation staining pattern after prolonged preincubation and a pyrophosphate system which produced a rapid change. Neutral salts in the preincubation medium accelerated the time course of the inactivation - activation process with the order for the halogen salts of K+ being F- less than Cl- less than Br- less than I-, which is a progression by molecular weight. The only sequence for cations on the myofibrillar actomyosin ATPase was Li+ less than Na+ less than K+. The response to salts was concentration dependent. An interaction existed between buffering compound, type of salt, and pH. These experiments demonstrate that the histochemical differentiation of fiber types by the myofibrillar actomyosin ATPase reaction depends upon a modification of some component(s) of the myofibrillar complex that can be influenced by a number of factors.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D002021:Buffers; D005260:Female; D006651:Histocytochemistry / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007477:Ions; D008297:Male; D009210:Myofibrils / Q000201:enzymology*; D009994:Osmolar Concentration; D051381:Rats; D012492:Salts
D002021:Buffers; D007477:Ions; D012492:Salts; D000251:Adenosine Triphosphatases