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用放射自显影法研究3H-地塞米松被大鼠垂体经免疫细胞化学鉴定的细胞摄取的情况。

Autoradiographic studies of 3H-dexamethasone uptake by immunocytochemically characterized cells of the rat pituitary.

作者信息

Rees H D, Stumpf W E, Sar M, Petrusz P

出版信息

Cell Tissue Res. 1977 Aug 26;182(3):347-56. doi: 10.1007/BF00219770.

Abstract

3H-Dexamethasone (10 microgram/kg) was injected intravenously in adrenalectomized rats and after survival times of 5, 30, 60, and 180 min its uptake within the pituitary was studied by autoradiography. Radioactivity was concentrated in cell nuclei in the pars nervosa and pars distalis. Within the pars intermedia, only cells of the marginal zone were labeled. In the pars distalis, some cells showed a weak nuclear accumulation of radioactivity as early as 5 min after injection. The tissue radioactivity was nearly maximal at 5 min, and the proportion of radioactivity in nuclei reached a maximum of 60-70% by 30 min. In competition experiments, non-radioactive steroids (1 mg/kg) were injected 5 min before 3H-dexamethasone and sacrifice was 30 min later. Dexamethasone markedly diminished the nuclear accumulation in the pars distalis, but corticosterone and progesterone did not. In the pars nervosa, corticosterone and progesterone competed for nuclear uptake of 3H-dexamethasone, although less effectively than dexamethasone itself. Different cell types in the pars distalis were characterized by treating autoradiograms with an immuno-peroxidase bridge procedure. Cells treated with anti-ACTH 17-39 had the greatest nuclear concentration of radioactivity, and those stained with anti-TSH were least heavily labeled. Cells treated with antisera to GH, PRL, and hCG were moderately labeled.

摘要

将3H-地塞米松(10微克/千克)静脉注射到肾上腺切除的大鼠体内,在5、30、60和180分钟的存活时间后,通过放射自显影研究其在垂体中的摄取情况。放射性集中在神经垂体和远侧部的细胞核中。在中间部,仅边缘区的细胞被标记。在远侧部,早在注射后5分钟,一些细胞就显示出放射性在细胞核中的微弱积累。组织放射性在5分钟时接近最大值,到30分钟时细胞核中的放射性比例达到60 - 70%的最大值。在竞争实验中,在注射3H-地塞米松前5分钟注射非放射性类固醇(1毫克/千克),30分钟后处死动物。地塞米松显著减少了远侧部细胞核中的积累,但皮质酮和孕酮没有。在神经垂体中,皮质酮和孕酮竞争3H-地塞米松的细胞核摄取,尽管效果不如地塞米松本身。通过免疫过氧化物酶桥法处理放射自显影片来鉴定远侧部不同的细胞类型。用抗促肾上腺皮质激素17 - 39处理的细胞放射性在细胞核中的浓度最高,用抗促甲状腺激素染色的细胞标记最少。用抗生长激素、抗催乳素和抗人绒毛膜促性腺激素抗血清处理的细胞标记程度适中。

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