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兔胃黏膜中的血管紧张素I转换酶:通过放射自显影、免疫荧光和免疫电子显微镜进行定位

Angiotensin I converting enzyme in gastric mucosa of the rabbit: localization by autoradiography, immunofluorescence, and immunoelectron microscopy.

作者信息

Laliberté F, Laliberté M F, Nonotte I, Bali J P, Chevillard C

机构信息

INSERM U.300, Faculté de Pharmacie, Montpellier, France.

出版信息

J Histochem Cytochem. 1991 Nov;39(11):1519-29. doi: 10.1177/39.11.1655876.

Abstract

To localize angiotensin converting enzyme (ACE) in the fundic mucosa of the rabbit, we used autoradiography with the ACE inhibitor [3H]-trandolaprilate and post-embedding immunocytochemical techniques with a goat anti-rabbit ACE, using fluorescence and electron microscopy. Autoradiographic localization of [3H]-trandolaprilate in rabbit fundus sections shows that ACE is present in the fundic mucosa and mainly in the gland area. Fundic mucosa was fixed with 4% formaldehyde and embedded in Lowicryl K4M. Semi-thin (1 micron) or thin sections (800-900 A) were stained with anti-rabbit ACE followed by fluorescein isothyocyanate-labeled rabbit anti-goat IgG or protein A-gold reagent, respectively. Label was present on endothelium of all blood vessels running through the mucosa. Label was prominently localized in the granules of mucous surface and neck cells and on the granules of chief cells. The intracellular localization of ACE, and particularly its intragranular presence within chief and mucous cells, suggests that the enzyme, at the fundic level, is involved in the intragranular processing of a peptide, the nature of which remains to be determined.

摘要

为了在兔胃底黏膜中定位血管紧张素转换酶(ACE),我们使用了[3H] - 群多普利拉(一种ACE抑制剂)进行放射自显影,并采用了用山羊抗兔ACE的包埋后免疫细胞化学技术,同时使用了荧光显微镜和电子显微镜。[3H] - 群多普利拉在兔胃底切片中的放射自显影定位显示,ACE存在于胃底黏膜中,主要在腺体区域。胃底黏膜用4%甲醛固定,然后包埋于Lowicryl K4M中。半薄切片(1微米)或超薄切片(800 - 900埃)分别用抗兔ACE染色,随后依次用异硫氰酸荧光素标记的兔抗山羊IgG或蛋白A - 金试剂处理。标记物出现在贯穿黏膜的所有血管的内皮上。标记物显著定位于黏液表面细胞和颈细胞的颗粒以及主细胞的颗粒上。ACE的细胞内定位,尤其是其在主细胞和黏液细胞颗粒内的存在,表明在胃底水平该酶参与了一种肽的颗粒内加工过程,其性质仍有待确定。

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