Mena F, Clapp C, Aguayo D, Martínez-Escalera G
Departamento de Fisiologia, Universidad Nacional Autónoma de México, México, D.F.
Neuroendocrinology. 1989 Feb;49(2):207-14. doi: 10.1159/000125116.
We studied the basal and thyrotropin-releasing hormone (TRH)-stimulated in vitro release of newly synthesized and mature prolactin (PRL) by lactating rat pituitary fragments. Newly synthesized PRL was defined as that resulting from either in vivo pulse labelling with [3H]-leucine (injected i.v. 10 min before removing the pituitary glands for in vitro incubation); or in vitro labelled by a 5-min preincubation period of adenohypophyseal fragments. Mature PRL was defined as that labelled in vivo 8 h before incubation. Medium PRL was quantified by polyacrylamide-gel-electrophoresis densitometric and liquid scintillation techniques. Under basal conditions, both total (unlabelled) and 8-hour labelled (mature) PRLs showed an in vitro release pattern characterized by an initial period of faster and higher secretion that lasted for 30-60 min followed by a lower secretion rate that extended from 60 to 240 min of incubation. Secretion of newly synthesized PRLs was detected since the first 30 min, but did not reach maximum levels until 90-120 min of incubation. Accordingly, PRL release during the first 30-60 min of incubation consisted primarily of mature PRL, whereas secretion during the rest of incubation included the release of newly synthesized hormone. These results confirm previous data on the sequential, rather than preferential, release of different age PRLs. TRH (0.35 microM) stimulated (about 30% above control) the release of both total and mature PRLs. No effect could be detected upon the release of in vivo labelled new PRL, while the same dose of TRH strongly stimulated (about 100% above control) the release of in vitro labelled new PRL.(ABSTRACT TRUNCATED AT 250 WORDS)
我们研究了泌乳大鼠垂体片段基础状态下以及促甲状腺激素释放激素(TRH)刺激后新合成的和成熟的催乳素(PRL)的体外释放情况。新合成的PRL定义为通过[3H] - 亮氨酸体内脉冲标记(在取出垂体进行体外孵育前10分钟静脉注射)产生的PRL;或通过腺垂体片段5分钟的预孵育期进行体外标记产生的PRL。成熟的PRL定义为孵育前8小时体内标记的PRL。通过聚丙烯酰胺凝胶电泳光密度法和液体闪烁技术对培养基中的PRL进行定量。在基础条件下,总(未标记)PRL和8小时标记(成熟)PRL均呈现出体外释放模式,其特征为最初30 - 60分钟分泌速度较快且量较高,随后分泌速率降低,持续至孵育60 - 240分钟。新合成的PRL在孵育最初30分钟即可检测到分泌,但直到孵育90 - 120分钟才达到最高水平。因此,孵育最初30 - 60分钟期间的PRL释放主要由成熟PRL组成,而孵育其余时间的分泌则包括新合成激素的释放。这些结果证实了先前关于不同年龄PRL顺序而非优先释放的数据。TRH(0.35微摩尔)刺激总PRL和成熟PRL的释放(比对照高约30%)。未检测到对体内标记的新PRL释放有影响,而相同剂量的TRH强烈刺激体外标记的新PRL释放(比对照高约100%)。(摘要截短于250字)