H+-adenosine triphosphatase (H+-ATPase) activity was demonstrated oytoohemioally in autophagio vaouoles (AVs) of rat hepatooytes using a modification of the method for the demonstration of neutral p-nitrophenyl phosph...H+-adenosine triphosphatase (H+-ATPase) activity was demonstrated oytoohemioally in autophagio vaouoles (AVs) of rat hepatooytes using a modification of the method for the demonstration of neutral p-nitrophenyl phosphatase (p-NPPase) activity [1]. When an inhibitor of H+-ATPase, N-ethylmaleimide (NEM) or 4,4'-diisothiooyanostilbene-2,2'disalfonio aold, di-sodium salt (BIDS) was included in the incubation medium the enyzme activity was abolished indicating that p-NPPase demonstrated in this study represents H+-ATPase. Autophagy was induced by a single intraperitoneal injection of vinblastine sulfate (VBL). The number of AVs increased remarkably in hepatooytes from 40 min after VBL treatment. H+-ATPase activity was observed mainly on the membranes of lysosomes and AVs. However, early forms of AVs containing only incompletely digested material showed no H+-ATPase activity. Most AVs revealing a positive reaction seemed to be in advanced stages of development. Acid phosphatase aotioity was demonstrable in mature but not in early forms of AVs. The present investigation showed that membranes of advanced stage A Vs possess an H+-ATPase which may be derived from lysosomal membranes.展开更多
文摘H+-adenosine triphosphatase (H+-ATPase) activity was demonstrated oytoohemioally in autophagio vaouoles (AVs) of rat hepatooytes using a modification of the method for the demonstration of neutral p-nitrophenyl phosphatase (p-NPPase) activity [1]. When an inhibitor of H+-ATPase, N-ethylmaleimide (NEM) or 4,4'-diisothiooyanostilbene-2,2'disalfonio aold, di-sodium salt (BIDS) was included in the incubation medium the enyzme activity was abolished indicating that p-NPPase demonstrated in this study represents H+-ATPase. Autophagy was induced by a single intraperitoneal injection of vinblastine sulfate (VBL). The number of AVs increased remarkably in hepatooytes from 40 min after VBL treatment. H+-ATPase activity was observed mainly on the membranes of lysosomes and AVs. However, early forms of AVs containing only incompletely digested material showed no H+-ATPase activity. Most AVs revealing a positive reaction seemed to be in advanced stages of development. Acid phosphatase aotioity was demonstrable in mature but not in early forms of AVs. The present investigation showed that membranes of advanced stage A Vs possess an H+-ATPase which may be derived from lysosomal membranes.
