Scale of autophagy estimated through lysosomal accumulation of L-lactate dehydrogenase in the liver of leupeptin-treated starved or refed rats
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The scale of autophagy in the liver of starved or refed rats was estimated by measuring the rate at which L-lactate dehydrogenase(LDH)accumulated in lysosomes after injecting a cysteine proteinase inhibitor, leupeptin, in vivo. The maximum accumulation rate of LDH in lysosomes was obtained from the differences in the lysosomal levels of LDH between 30 and 45 min after leupeptin injection in the rats starved overnight and was estimated to be 2.1% of total liver LDH per hour. In the rats refed immediately after injecting leupeptin into starved rats, the accumulation of LDH into lysosomes was almost completely stopped 30 min after refeeding. Considering that leupeptin-induced suppression of intralysosomal degradation of native LDH was almost complete but that of acid-denatured LDH was incomplete, it is cocluded that the scale of autophagy is 2.1% or more of total cellular LDH per hour in the liver of starved rat in vivo and that autophagy enhanced by starvation is almost completely suppressed 30 min after refeeding.