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KMID : 0311119780190020001
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Yonsei Medical Journal
1978 Volume.19 No. 2 p.1 ~ p.10
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Electron Microscopy on Activity and Localization of Glucose-6-phosphatase in Liver Cells
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Shin, Tai Sun
Chung, In Hyuk/Kim, Soo Sung
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Abstract
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It is interesting and in important to study histochemical changes of glucose-6-phosphatase(G-6-Pase) activity by electron microscopy in order to promote the knowledge needed for diagnosis and prognosis in such liver diseases as von Gierke´s disease, hepatoma and various other hepatocellular alterations of different origins. Since we had not accomplished the electron microscopic demonstration of G-6-Pase, although light microscopic studies on changes of the enzyme activity were done in this laboratory, this investigation was planned to obtain a satisfactory technique for ultrastructural demonstration of the enzyme activity.
Unfixed frozen sections(80 u thick) of mouse liver were washed for 2~3 minutes in a 0.4 M sucrose solution (pH6.8) containing 4 mM lead nitrate and then incubated for 15~20 minutes at 32~37℃ in several different media to which 0.4M sucrose solution was added: A) a modification of the original Chiquoine medium, B) the first modification of the Wachstein-Meisel medium C(the second modification; the 2% lead nitrate solution was reduced in amount to 1.5 ㎖ instead of 3.0 ㎖ in the medium-B). After incubation, these sections were fixed in 1% osmic acid containing sucrose, followed by embedding in Epon, ultrathin-section, mounting and staining with uranyl acetate and/or lead nitrate. By incubating the sections in the medium (B or C), satisfactory preparations were obtainable for its electron microscopic demonstration. The granular deposits of reaction products were found characteristically on the membranous component of the rough-and smooth-surfaced endoplasmic reticulum and unclear envelope.
Occasional deposits were observed within cisternae or vesicles, in the nucleus, and immediately adjacent to the cisternal membrane and glycogen areas.
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