MacGregor R R, Hamilton J W, Cohn D V
Endocrinology. 1975 Jul;97(1):178-88. doi: 10.1210/endo-97-1-178.
Bovine parathyroid gland slices were incubated in Krebs' buffer with [3-H] leucine in order to assess the biosynthesis and secretion of parathyroid hormone (PTH). After incubation the particulate structures of the tissue were extracted with sodium deoxycholate to yield primarily newly synthesized (radioactive) PTH in the extract and preexisting granule PTH in the residue. In pulse-chase experiments radioactive PTH entered the granule fraction at a time when total tissue radioactive hormone was declining, which indicates that some newly-synthesized PTH was packaged into these granules. The specific radioactivity of the PTH in the incubation medium was from 20-fold to more than 80-fold greater than that of the granule PTH of the tissue, but was similar in magnitude to that of the tissue's newly synthesized hormone. Secretion of newly synthesized PTH was greater during incubation in buffer with 1.25 mM Ca than it was at 2.5 mM Ca which indicated that the hormone release was subject to physiological control. The PTH content of the tissue granule fraction was lower following incubation at 1.25 mM Ca than at 2.5 mM Ca. The possibility was excluded that the higher specific radioactivity of PT1 of incubation medium compared to that of the secretory granule fraction resulted from a diffusion gradient of [3-H] leucine into the tissue slices. These data indicate that a major portion of the newly synthesized PTH was secreted without prior equilibration with the hormone in the pool of secretory granules.
将牛甲状旁腺切片置于含有[3-H]亮氨酸的 Krebs 缓冲液中孵育,以评估甲状旁腺激素(PTH)的生物合成和分泌。孵育后,用脱氧胆酸钠提取组织的颗粒结构,提取物中主要产生新合成的(放射性)PTH,残渣中则含有预先存在的颗粒状 PTH。在脉冲追踪实验中,当总组织放射性激素下降时,放射性 PTH 进入颗粒部分,这表明一些新合成的 PTH 被包装到这些颗粒中。孵育培养基中 PTH 的比放射性比组织颗粒状 PTH 的比放射性高 20 倍至 80 倍以上,但与组织新合成激素的比放射性大小相似。在含有 1.25 mM 钙的缓冲液中孵育期间,新合成 PTH 的分泌量大于在 2.5 mM 钙时的分泌量,这表明激素释放受生理控制。在 1.25 mM 钙条件下孵育后,组织颗粒部分的 PTH 含量低于在 2.5 mM 钙时的含量。排除了孵育培养基中 PTH 的比放射性高于分泌颗粒部分的比放射性是由于[3-H]亮氨酸向组织切片扩散梯度所致的可能性。这些数据表明,新合成的 PTH 的大部分在没有与分泌颗粒池中的激素预先平衡的情况下就被分泌了。