Cardell R R, Michaels J E, Hung J T, Cardell E L
J Cell Biol. 1985 Jul;101(1):201-6. doi: 10.1083/jcb.101.1.201.
Hormonal control of hepatic glycogen and blood glucose levels is one of the major homeostatic mechanisms in mammals: glycogen is synthesized when portal glucose concentration is sufficiently elevated and degraded when glucose levels are low. We have studied initial events of hepatic glycogen synthesis by injecting the synthetic glucocorticoid dexamethasone (DEX) into adrenalectomized rats fasted overnight. Hepatic glycogen levels are very low in adrenalectomized rats, and DEX causes rapid deposition of the complex carbohydrate. Investigation of the process of glycogen deposition was performed by light and electron microscopic (EM) radioautography using [3H]galactose as a glycogen precursor. Rats injected with DEX for 2-3 h and [3H]galactose one hour before being killed displayed an increasing number of intensely labeled hepatocytes. EM radioautography revealed silver grains over small (+/- 1 micron) ovoid or round areas of the cytosome that were rich in smooth endoplasmic reticulum (SER) and contained a high concentration of small dense particles. These distinct areas or foci of SER and presumptive glycogen (SERGE) were most numerous during initial periods of glycogen synthesis. After longer exposure to DEX (4-5 h) more typical deposits of cytoplasmic glycogen were evident in the SERGE regions. Several criteria indicated that the SERGE foci contained glycogen or presumptive glycogen: resemblance of the largest dense particles to beta-glycogen particles in EM; association of 3H-carbohydrate with the foci; removal of particles and label with alpha-amylase; and positive reaction with periodic acid-chromic acid-silver methenamine. The concentration of SER in the small foci and the association of newly formed glycogen particles with elements of SER suggest a role for this organelle in the initial synthesis of glycogen.
当门静脉葡萄糖浓度充分升高时,糖原被合成;当葡萄糖水平较低时,糖原被分解。我们通过向过夜禁食的肾上腺切除大鼠注射合成糖皮质激素地塞米松(DEX),研究了肝糖原合成的起始事件。肾上腺切除大鼠的肝糖原水平非常低,而DEX会导致复合碳水化合物的快速沉积。使用[3H]半乳糖作为糖原前体,通过光镜和电镜放射自显影对糖原沉积过程进行了研究。在处死前2 - 3小时注射DEX并在处死前1小时注射[3H]半乳糖的大鼠,显示出标记强烈的肝细胞数量增加。电镜放射自显影显示,在富含滑面内质网(SER)且含有高浓度小致密颗粒的细胞质小(±1微米)卵圆形或圆形区域上有银颗粒。这些SER和假定糖原的独特区域或灶(SERGE)在糖原合成的初始阶段最为丰富。在长时间暴露于DEX(4 - 5小时)后,SERGE区域出现了更典型的细胞质糖原沉积物。几个标准表明SERGE灶含有糖原或假定糖原:电镜下最大致密颗粒与β-糖原颗粒相似;3H-碳水化合物与灶的关联;用α-淀粉酶去除颗粒和标记;以及与高碘酸-铬酸-亚甲胺银的阳性反应。小灶中SER的浓度以及新形成的糖原颗粒与SER成分的关联表明该细胞器在糖原的初始合成中起作用。