Konoplytska O L, Sytnyanska N P
Ukr Biokhim Zh. 1975 Nov-Dec;47(6):769-75.
The optimal conditions are selected for electron-cytochemical detection of the ATPase activity in nuclei of the skeletal muscles of rabbits and nuclei of Vicia faba L. meristem. It is shown that the previous fixation of nuclei in the rabbit skeletal muscle for 10 min in a mixture of the buffer solutions of 4% glutaric dialdehyde and 4% neutral formalin (1:1) causes a decrease in their ATPase activity by 78% in the medium containing Mg2+ and by 34% - in the medium containing Ca2+; in nuclei of horse bean seedlings meristem it lowers respectively by 28 and 16%. Ions of lead in a concentration of 0.4 mM evoke a decrease in the ATPase activity in the medium containing Mg2+, in nuclei of the rabbit skeletal muscles by 35% and in nuclei of horse bean meristem by 15% in the medium containing Ca2+. The vaule of the residual activity is sufficient for detection of the product of ATP enzymic hydrolysis reaction by activity is sufficient for detection of the product of ATP enzymic hydrolysis reaction by the method of electronic cytochemistry. An increase in the Pb2+ concentration higher than 2.8 mM evokes nonenzymic hydrolysis of ATP. The ATPase activity under the electron-cytochemical study is found within the range of pH 6.3-8.5. The product of reaction forms most intensively at pH 7.2-7.5 in the medium with both Mg2+ and Ca2+.
选择了用于兔骨骼肌细胞核和蚕豆分生组织细胞核中ATP酶活性电子细胞化学检测的最佳条件。结果表明，兔骨骼肌细胞核预先在4%戊二醛和4%中性福尔马林（1:1）的缓冲液混合物中固定10分钟，在含Mg2+的培养基中其ATP酶活性降低78%，在含Ca2+的培养基中降低34%；在蚕豆幼苗分生组织细胞核中分别降低28%和16%。浓度为0.4 mM的铅离子在含Mg2+的培养基中使兔骨骼肌细胞核的ATP酶活性降低35%，在含Ca2+的培养基中使蚕豆分生组织细胞核的ATP酶活性降低15%。残余活性值足以通过电子细胞化学方法检测ATP酶水解反应的产物。Pb2+浓度高于2.8 mM时会引发ATP的非酶水解。电子细胞化学研究中ATP酶活性在pH 6.3 - 8.5范围内被发现。反应产物在含Mg2+和Ca2+的培养基中于pH 7.2 - 7.5时形成最为强烈。
