Kuriakose N R, Reifel C W, Bendayan M, Elce J S, Shin S H
Department of Anatomy, Queen's University, Kingston, Ontario, Canada.
Histochemistry. 1989;92(6):499-503. doi: 10.1007/BF00524761.
The phenomenon of crinophagy in rat pituitary mammotrophs, or lysosomal uptake of prolactin secretory granules, was confirmed by means of double-label immunogold electron microscopy, and shown to be induced in estrogen-stimulated male rats. Rabbit antibodies to rat cathepsin D were used to label lysosomes, and to rat prolactin to label secretory granules. The pituitaries were fixed in 4% formaldehyde and 1% glutaraldehyde, embedded in Lowicryl K4M, and thin sections were exposed successively to primary antibodies, biotin-labelled second antibodies, and streptavidin-gold, with an amplification procedure for cathepsin D. Cathepsin D and prolactin were detected separately on opposite sides of the sections, using 5-nm and 15-nm gold particles. Lysosomal uptake of prolactin secretory granules was not observed in untreated control rats. It was detected in about 26% of lysosome-containing mammotroph cell sections in estrogen-stimulated rats and at 7 h after estrogen withdrawal, but fell to 14% at 24 h and to 2% at 72 h after estrogen withdrawal.
通过双标记免疫金电子显微镜技术证实了大鼠垂体催乳素细胞中的自噬现象,即催乳素分泌颗粒的溶酶体摄取,并表明在雌激素刺激的雄性大鼠中会诱导这种现象。用兔抗大鼠组织蛋白酶D抗体标记溶酶体,用兔抗大鼠催乳素抗体标记分泌颗粒。垂体用4%甲醛和1%戊二醛固定,包埋在Lowicryl K4M中,超薄切片依次与一抗、生物素标记的二抗和链霉亲和素金反应,并对组织蛋白酶D进行放大处理。使用5纳米和15纳米的金颗粒在切片的相对两侧分别检测组织蛋白酶D和催乳素。在未处理的对照大鼠中未观察到催乳素分泌颗粒的溶酶体摄取。在雌激素刺激的大鼠中,约26%含有溶酶体的催乳素细胞切片以及雌激素撤药后7小时检测到这种现象,但在雌激素撤药后24小时降至14%,72小时降至2%。