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凝集素标记囊泡在肝癌腹水细胞GERL区的细胞化学定位

Cytochemical localization of lectin labeled vesicles in GERL region of hepatoma ascites cells.

作者信息

Moller P C, Wang J J, Yokoyama M, Chang J P

出版信息

Histochemistry. 1979 Aug;62(3):289-97. doi: 10.1007/BF00508357.

Abstract

The fate of lectin labeled internalized plasma membrane in the ascites tumor form of the Chang rat hepatoma growing under in vivo and in vitro conditions was investigated cytochemically. Ascites cells were incubated in Convanavalin A (Con A) and horseradish peroxidase (PO), either with or without prior glutaraldehyde fixation and subsequently treated with 3',3-diaminobenzidine. In cells fixed before Con-A-PO labeling the reaction product was localized as a continuous and even layer upon the external surface of the plasma membrane. If unfixed cells were treated with Con A, coupled with PO at 4 degrees C and reincubated in phosphate buffered saline at 37 degrees C for varying periods of time, the Con-A-PO layer was of irregular thickness. In as little as 15 min of reincubation endocytotic vesicles containing PO positive material were closely associated with GERL components of the Golgi Apparatus. Localization of acid phosphatase (ACPase) within GERL vesicles, similar in size and location to those containing Con-A-PO reaction product, indicates that the Con-A-PO labeled vesicles may be a component of the Golgi apparatus in hepatoma cells.

摘要

采用细胞化学方法研究了在体内和体外条件下生长的张氏大鼠肝癌腹水瘤形式中,凝集素标记的内化质膜的命运。腹水细胞在伴刀豆球蛋白A(Con A)和辣根过氧化物酶(PO)中孵育,有无预先戊二醛固定,随后用3',3-二氨基联苯胺处理。在Con-A-PO标记前固定的细胞中,反应产物定位在质膜外表面,呈连续均匀的层。如果未固定的细胞在4℃用Con A和PO偶联处理,并在37℃的磷酸盐缓冲盐水中再孵育不同时间,Con-A-PO层厚度不规则。再孵育仅15分钟,含有PO阳性物质的内吞小泡就与高尔基体的GERL成分紧密相关。酸性磷酸酶(ACPase)在GERL小泡中的定位,其大小和位置与含有Con-A-PO反应产物的小泡相似,表明Con-A-PO标记的小泡可能是肝癌细胞中高尔基体的一个组成部分。

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