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采用包埋前和包埋后免疫细胞化学方法对去势猪垂体细胞中性腺促激素的亚细胞定位研究。

Subcellular localization of gonadotropic hormones in pituitary cells of the castrated pig with the use of pre- and post-embedding immunocytochemical methods.

作者信息

Dacheux F

出版信息

Cell Tissue Res. 1984;236(1):153-60. doi: 10.1007/BF00216525.

Abstract

Pre- and post-embedding immunocytochemical methods based on the use of specific antibodies against beta-subunits of porcine LH and FSH were applied to determine the changes occurring in the anterior pituitary of the pig after gonadectomy. The results showed that (1) the total number of immunoreactive gonadotropes increased from 21-25% in control animals to 24-37% in castrated animals; (2) all gonadotropes contained both LH and FSH; (3) several types of immunoreactive LH/FSH cells were revealed; and (4) the two immunocytochemical methods used with dispersed cells localized the hormones in the same subcellular sites. However, the staining intensity in the different locations varied depending on the method applied. With the post-embedding method, a dense reaction product was found in the secretory granules but the cisternae of RER and the Golgi saccules were always slightly reactive. After the pre-embedding method, the staining intensity in the RER-cisternae and in the Golgi saccules was greatly increased. Thus, the two methodological approaches used in this study have permitted to visualize immunocytochemically the gonadotropic hormones not only at the sites of their storage but also along the intracellular pathway of the secretory material, i.e., at the site of its synthesis and during its passage via the Golgi zone.

摘要

基于使用针对猪促黄体生成素(LH)和促卵泡生成素(FSH)β亚基的特异性抗体的包埋前和包埋后免疫细胞化学方法,被用于确定去势后猪垂体前叶发生的变化。结果显示:(1)免疫反应性促性腺激素细胞的总数从对照动物的21%-25%增加到去势动物的24%-37%;(2)所有促性腺激素细胞都同时含有LH和FSH;(3)揭示了几种类型的免疫反应性LH/FSH细胞;(4)用于分散细胞的两种免疫细胞化学方法将激素定位在相同的亚细胞部位。然而,不同部位的染色强度因所应用的方法而异。采用包埋后方法时,在分泌颗粒中发现密集的反应产物,但粗面内质网池和高尔基体囊泡总是有轻微反应。采用包埋前方法后,粗面内质网池和高尔基体囊泡中的染色强度大大增加。因此,本研究中使用的两种方法不仅能够在促性腺激素的储存部位,而且能够沿着分泌物质的细胞内途径,即在其合成部位和通过高尔基体区域的过程中,通过免疫细胞化学方法可视化促性腺激素。

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