المؤلفون: Paul B. Gordon, Per Ottar Seglen

المصدر: Biochemical and biophysical research communications. 151(1)

مصطلحات موضوعية: Male, Endocytic cycle, Biophysics, Lactose, Vacuole, Biology, Vinblastine, Biochemistry, chemistry.chemical_compound, Phagocytosis, Autophagy, Animals, Asparagine, Molecular Biology, Cells, Cultured, chemistry.chemical_classification, Rats, Inbred Strains, Cell Biology, Compartment (chemistry), beta-Galactosidase, Endocytosis, Cell biology, Rats, Cytosol, Enzyme, chemistry, Liver, Vacuoles, Lysosomes

الوصف: [14C]Lactose, electroinjected into the cytosol of isolated rat hepatocytes, was sequestered by autophagy, transferred to lysosomes and eventually hydrolysed. Asparagine prevented the fusion between prelysosomal autophagic vacuoles and lysosomes, and caused lactose to accumulate in the former. However, if the hepatocytes were simultaneously allowed to endocytose added β-galactosidase, no lactose accumulation occurred. These results suggest that autophagically sequestered lactose and endocytosed β-galactosidase were delivered to the same prelysosomal vacuole, where the lactose was hydrolysed by the enzyme. The name amphisome is suggested for this new functional compartment, common to the autophagic and endocytic pathways.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a5669d65cf3dfb38c3cff5f8e20b7dedTest
https://pubmed.ncbi.nlm.nih.gov/3126737Test

المؤلفون: Per Ottar Seglen, H. Hoyvik, P. B. Gordon

المصدر: Experimental cell research. 166(1)

مصطلحات موضوعية: Male, Sucrose, Density gradient, Lactose, Vacuole, Biology, Vinblastine, chemistry.chemical_compound, Phagocytosis, Lysosome, Phagosomes, Organelle, medicine, Autophagy, Animals, Phagosome, Adenine, Hydrolysis, Chloroquine, Rats, Inbred Strains, Cell Biology, Rats, Cytosol, medicine.anatomical_structure, Biochemistry, chemistry, Liver, Lysosomes

الوصف: [14C]Lactose, introduced into the cytosol of isolated rat hepatocytes by means of electropermeabilization, is sequestered autophagically in the same way as the established sequestration probe, [14C]sucrose. However, unlike the inert sucrose molecule, lactose is rapidly hydrolysed in the lysosomes, and can therefore be used to probe the last step of the autophagic pathway (i.e. fusion with the lysosome). During autophagy lactose is present only at a low, steady-state level in pre-lysosomal vacuoles (probably autophagosomes), serving as a useful marker for these organelles. If autophagosome-lysosome fusion is blocked with vinblastine (Kovacs et al., Exp cell res 137 (1982) 191), [14C]lactose will accumulate continuously as a function of the sequestration rate, and reach a high level in the pre-lysosomal vacuoles. Density gradient analysis, using chloroquine (CLQ) to alter lysosomal density, suggests that these organelles have a broad density distribution (1.08-1.13 g/ml), thus differing significantly from the distribution of lysosomes.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::c46f164170981e9a73cc29b88622bbe1Test
https://pubmed.ncbi.nlm.nih.gov/3743649Test