CHINESE-HAMSTER OVARY CELL LYSOSOMES RAPIDLY EXCHANGE CONTENTS
CHINESE-HAMSTER OVARY CELL LYSOSOMES RAPIDLY EXCHANGE CONTENTS
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DOI：
10.1083/jcb.105.6.2703
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发表时间：
1987-12-01
影响因子：
7.8
通讯作者：
STORRIE, B
中科院分区：
文献类型：
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作者：
FERRIS, AL;BROWN, JC;STORRIE, B
我们已经使用细胞融合来解决大分子是否在溶酶体之间快速交换的问题。供体细胞溶酶体通过液体相胞饮标记物、转糖酶(蔗糖酶)、荧光黄、FITC标记的葡聚糖或德克萨斯红标记的葡聚糖长期内化来标记。受体细胞含有因长期内化稀糖而肿胀的溶酶体或以FITC-葡聚糖摄取为标志。在细胞融合之前，细胞在无标记的培养液中孵育1或2小时，以清除内膜室中的任何标记。受体细胞感染泡状口炎病毒作为FusoGen。供体细胞和受体细胞共培养1或2小时，然后在短暂的pH 5中融合。在所有病例中，供体和受体细胞溶酶体在17℃下广泛交换内容。细胞合胞在17℃孵育。C抑制了溶酶体/溶酶体交换，但在17℃时似乎发生了“启动”过程。C.含有蔗糖/阳性溶酶体的细胞间融合中溶酶体/溶酶体交换的动力学表明，溶酶体/溶酶体交换即使不是比内切酶/溶酶体交换更快，也是一样快的。这些实验表明，在体内，溶酶体是一个快速混合的细胞器隔室。
We have used cell fusion to address the question of whether macromolecules are rapidly exchanged between lysosomes. Donor cell lysosomes were labeled by the long-term internalization of the fluid-phase pinocytic markers, invertase (sucrase), Lucifer Yellow, FITC-conjugated dextran, or Texas red-conjugated dextran. Recipient cells contained lysosomes swollen by long-term internalization of dilute sucrose or marked by an overnight FITC-dextran uptake. Cells were incubated for 1 or 2 h in marker-free media before cell fusion to clear any marker from an endosomal compartment. Recipient cells were infected with vescicular stomatitis virus as a fusogen. Donor and recipient cells were co-cultured or 1 or 2 h and then fused by a brief exposure to pH 5. In all cases, extensive exchange of content between donor and recipient cell lysosomes was observed at 17.degree.C. Incubation of cell syncytia at 17.degree. C blocked lysosome/lysosome exchange, although a "priming" process(es) appeared to occur at 17.degree.C. The kinetics of lyso-some/lysosome exchange in fusions between cells containing invertase-positive lysosomes and sucrose/positive lysosomes indicated that lysosome/lysosome exchange was as rapid, if not more rapid, than endosome/lysosome exchange. These experiments suggest that in vivo the lysosome is a rapidly intermixing organellar compartment.