Michaels J E, Cardell R R
Department of Anatomy and Cell Biology, University of Cincinnati College of Medicine, Ohio 45267-0521.
Anat Rec. 1993 Jul;236(3):486-92. doi: 10.1002/ar.1092360309.
Hepatic glycogen synthase activity was localized in normal and adrenalectomized (ADX) rats after fasting overnight and in fasted ADX rats after injection of dexamethasone (DEX) 2-8 h prior to sacrifice to stimulate glycogen synthesis. Cryostat sections were incubated in medium containing substrate to demonstrate glycogen synthase activity as indicated by glycogen synthesized during incubation. Sections from fasted normal rats showed limited dispersed glycogen synthase activity in both periportal and centrilobular regions. In contrast, activity for glycogen synthase in hepatocytes from fasted ADX rats appeared as large aggregates in random hepatocytes throughout the lobule. Two hours after injection of DEX the reaction product appeared as aggregates in some hepatocytes, but other cells revealed dispersed enzyme activity. Glycogen synthase activity was evident in more hepatocytes after 4 h treatment with DEX and after 8 h virtually all hepatocytes contained abundant reaction product. The results suggest that synthase activity becomes concentrated in limited regions of selected hepatocytes in fasted ADX rats. DEX stimulation of glycogen synthesis for 4-8 h results in increased enzyme activity.
在过夜禁食后的正常大鼠和肾上腺切除(ADX)大鼠中,以及在处死前2 - 8小时注射地塞米松(DEX)以刺激糖原合成的禁食ADX大鼠中,对肝糖原合酶活性进行定位。将冰冻切片在含有底物的培养基中孵育,以通过孵育期间合成的糖原所指示的情况来证明糖原合酶活性。禁食正常大鼠的切片在门周和小叶中央区域均显示出有限的分散的糖原合酶活性。相比之下,禁食ADX大鼠肝细胞中的糖原合酶活性表现为整个小叶中随机分布的肝细胞内有大的聚集物。注射DEX两小时后,反应产物在一些肝细胞中表现为聚集物,但其他细胞显示出分散的酶活性。用DEX处理4小时后,更多肝细胞中糖原合酶活性明显,8小时后几乎所有肝细胞都含有丰富的反应产物。结果表明,在禁食的ADX大鼠中,合酶活性集中在选定肝细胞的有限区域。DEX刺激糖原合成4 - 8小时会导致酶活性增加。