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雄性、雌性和哺乳期雌性大鼠腺垂体特定细胞中催乳素和生长激素的共定位

Colocalization of prolactin and growth hormone within specific adenohypophyseal cells in male, female, and lactating female rats.

作者信息

Nikitovitch-Winer M B, Atkin J, Maley B E

出版信息

Endocrinology. 1987 Aug;121(2):625-30. doi: 10.1210/endo-121-2-625.

Abstract

The localization of PRL and GH within adenohypophysial cells has been investigated with immunocytochemical methodology using colloidal gold of different sizes. Classically, using morphological criteria at the light and electron microscope levels, two types of individual cells have been described which, it is believed, exclusively produce either PRL or GH, i.e. mammotrophs and somatotrophs. Since it has been reported that some gonadotrophs may colocalize and secrete both FSH and LH, and that some unidentified cells release both PRL and GH, we designed a study to investigate whether PRL and GH are present only in their respective specific cells or may be colocalized in mammotrophs and somatotrophs or possibly other types of cells within the pars distalis. Using immunocytochemistry at the electron microscope level we were able to label the two hormones with different sizes of colloidal gold bound to a second antibody and visualize them within pituitary cells. Two different primary antibodies to PRL and GH as well as a more purified antibody to GH were used. Pituitaries from cycling and lactating female rats and adult males were processed appropriately for electron microscopic studies. After sectioning, individual grids were treated with either primary antibodies for PRL or GH, or the same grid was treated sequentially with these two antibodies. All primary antibodies were absorbed with the heterologous hormone before usage. The second antibodies were bound to colloidal gold particles of either 10 nm (for GH) or 20 nm (for PRL) diameter, so that selective visualization of the two hormones could be achieved within the same cells. It was observed that mammotrophs immunolabeled only for PRL, and somatotrophs labeled only for GH regardless of the source of the antibody. However, an atypical small granule cell, a possible mammosomatotroph, colabeled consistently for both PRL and GH in all types of animals used and with antibodies from all of above sources. This report gives for the first time morphological evidence for the existence of pituitary cells that colocalize both PRL and GH in the normal rat pituitary gland. The possibility that these bihormonal cells represent stem cells which may give rise to both mammotrophs and somatotrophs under appropriate stimulation or that they may release both hormones under the influence of unspecific stimuli is suggested.

摘要

采用不同大小的胶体金免疫细胞化学方法,对腺垂体细胞内催乳素(PRL)和生长激素(GH)的定位进行了研究。传统上,根据光镜和电镜水平的形态学标准,已描述了两种类型的单个细胞,据信它们分别专门产生PRL或GH,即催乳素细胞和生长激素细胞。由于有报道称一些促性腺激素细胞可能同时共定位并分泌促卵泡激素(FSH)和促黄体生成素(LH),且一些未鉴定的细胞可同时释放PRL和GH,因此我们设计了一项研究,以调查PRL和GH是否仅存在于各自特定的细胞中,或者是否可能在催乳素细胞和生长激素细胞或远侧部的其他类型细胞中共定位。利用电镜水平的免疫细胞化学技术,我们能够用与二抗结合的不同大小的胶体金标记这两种激素,并在垂体细胞内观察到它们。使用了两种针对PRL和GH的不同一抗以及一种更纯化的针对GH的抗体。对处于发情周期和哺乳期的雌性大鼠以及成年雄性大鼠的垂体进行适当处理,以进行电镜研究。切片后,将单个载网分别用PRL或GH的一抗处理,或者将同一载网依次用这两种抗体处理。所有一抗在使用前均用异源激素进行吸附。二抗与直径为10nm(用于GH)或20nm(用于PRL)的胶体金颗粒结合,以便在同一细胞内实现对这两种激素的选择性观察。观察到,无论抗体来源如何,催乳素细胞仅对PRL进行免疫标记,生长激素细胞仅对GH进行标记。然而,一种非典型的小颗粒细胞,可能是泌乳生长激素细胞,在所有使用的动物类型中以及使用上述所有来源的抗体时,均始终对PRL和GH进行共标记。本报告首次提供了正常大鼠垂体中存在同时共定位PRL和GH的垂体细胞的形态学证据。有人提出,这些双激素细胞可能代表干细胞,在适当刺激下可能产生催乳素细胞和生长激素细胞,或者在非特异性刺激的影响下可能释放两种激素。

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