Localization of glucose-6-phosphatase activity in the rat cardiac muscle by cobalt-based cytochemistry.
The histocytochemical method for the detection of glucose-6-phosphatase activity has been studied in the rat heart tissue by using cobalt-capture technique. The incubation medium had the following ingredients: cobalt chloride, PIPES-NaOH buffer pH = 6.7, glucose-6-phosphate disodium salt and phenylalanine. Cobalt was revealed by using a solution of 1% ammonium sulfide and was fixed by osmium tetroxide. The most effective concentration of cobalt chloride was 50 mM. With a concentration lower than 30 mM, the reaction products were not determined. With a higher concentration, a small non-specific precipitation was visualized. Ultracytochemically, a strong enzyme reaction was present in the nuclear envelopes of cardiac myocytes, endotheliocytes and in the sarcoplasmic reticulum. These results imply that the method used is valid and can be recommended for studying the localization of glucose-6-phosphatase activity.
Zinchuk
V S
VS
Department of Pathology, State Medical University, Kiev, Ukraine.
D000818:Animals; D002467:Cell Nucleus / Q000201:enzymology / Q000648:ultrastructure; D003035:Cobalt / Q000737:chemistry*; D005952:Glucose-6-Phosphatase / Q000378:metabolism*; D006651:Histocytochemistry; D008297:Male; D009132:Muscles / Q000201:enzymology / Q000648:ultrastructure; D009206:Myocardium / Q000201:enzymology* / Q000648:ultrastructure; D051381:Rats; D017208:Rats, Wistar; D012519:Sarcoplasmic Reticulum / Q000201:enzymology / Q000648:ultrastructure; D016707:Tissue Fixation
D003035:Cobalt; D005952:Glucose-6-Phosphatase