Saermark T, Jones P M, Robinson I C
Biochim Biophys Acta. 1985 Sep 25;819(1):114-8. doi: 10.1016/0005-2736(85)90201-9.
[3H]Choline and [35S]methionine injected into the guinea pig hypothalamus in vivo were incorporated into the lipids and proteins, respectively, of secretory vesicles transported to the neural lobe. Prolonged in vivo stimulation of hormone secretion by dehydration decreased the [3H]choline content of secretory vesicles, with a concomitant increase in the [3H]choline content of a membrane fraction isolated on sucrose gradients. After stimulation of neural lobes in vitro in the presence of horseradish peroxidase, this extracellular fluid marker was found in the same membrane fraction. SDS electrophoresis of membrane proteins radiolabelled by [35S]methionine in vivo demonstrated that this fraction contained at least one major protein also present in the secretory vesicle membrane. These results suggest that we have isolated a membrane fraction containing the structure(s) involve in membrane retrieval in the neurohypophysis.
体内注射到豚鼠下丘脑的[³H]胆碱和[³⁵S]甲硫氨酸分别被转运至神经叶的分泌囊泡的脂质和蛋白质中。脱水对激素分泌进行体内长期刺激会降低分泌囊泡的[³H]胆碱含量,同时蔗糖梯度分离的膜组分中的[³H]胆碱含量会增加。在辣根过氧化物酶存在的情况下对神经叶进行体外刺激后,这种细胞外液标记物出现在相同的膜组分中。体内用[³⁵S]甲硫氨酸进行放射性标记的膜蛋白的SDS电泳表明,该组分至少含有一种也存在于分泌囊泡膜中的主要蛋白质。这些结果表明,我们已经分离出了一个含有参与神经垂体膜回收结构的膜组分。