scholarly journals Effects of aminoglycoside antibiotics on lysosomal enzymes of rat kidney.

1983 ◽  
Vol 31 (9) ◽  
pp. 3277-3283 ◽  
Author(s):  
YUKIHIKO ARAMAKI ◽  
ASAICHI INABA ◽  
JUNJI NIIBUCHI ◽  
SEISHI TSUCHIYA
Keyword(s):  
Lysosomal Enzymes ◽  
Rat Kidney ◽  
Aminoglycoside Antibiotics

scholarly journals Distribution of newly synthesized lysosomal enzymes in the endocytic pathway of normal rat kidney cells.

1991 ◽  
Vol 115 (6) ◽  
pp. 1561-1572 ◽  
Author(s):  
T Ludwig ◽  
G Griffiths ◽  
B Hoflack
Keyword(s):  
Lysosomal Enzymes ◽  
Rat Kidney ◽  
Fluid Phase ◽  
Endocytic Pathway ◽  
Early Endosomes ◽  
Fluid Phase Endocytosis ◽  
Normal Rat ◽  
Endocytic Compartments ◽  
Endocytic Pathways

We have investigated the distribution of newly synthesized lysosomal enzymes in endocytic compartments of normal rat kidney (NRK) cells. The mannose-6-phosphate (Man6-P) containing lysosomal enzymes could be iodinated in situ after internalization of lactoperoxidase (LPO) by fluid phase endocytosis and isolated on CI-MPR affinity columns. For EM studies, the ectodomain of the CI-MPR conjugated to colloidal gold was used as a probe specific for the phosphomannosyl marker of the newly synthesized hydrolases. In NRK cells, approximately 20-40% of the phosphorylated hydrolases present in the entire pathway were found in early endocytic structures proximal to the 18 degrees C temperature block including early endosomes. These structures were characterized by a low content of endogenous CI-MPR and were accessible to fluid phase markers internalized for 5-15 min at 37 degrees C. The bulk of the phosphorylated lysosomal enzymes was found in late endocytic structures distal to the 18 degrees C block, rich in endogenous CI-MPR and accessible to endocytic markers internalized for 30-60 min at 37 degrees C. The CI-MPR negative lysosomes were devoid of phosphorylated hydrolases. This distribution was unchanged in cells treated with Man6-P to block recapture of secreted lysosomal enzymes. However, lysosomal enzymes were no longer detected in the early endosomal elements of cells treated with cycloheximide. Immunoprecipitation of cathepsin D from early endosomes of pulse-labeled cells showed that this hydrolase is a transient component of this compartment. These data indicate that in NRK cells, the earliest point of convergence of the lysosomal biosynthetic and the endocytic pathways is the early endosome.

scholarly journals Release of enzymes from the lysosomes of rat kidney cortex by aminoglycoside antibiotics.

1984 ◽  
Vol 32 (8) ◽  
pp. 3173-3178 ◽  
Author(s):  
ASAICHI INABA ◽  
YUKIHIKO ARAMAKI ◽  
KAZUHISA TAKEDA ◽  
SEISHI TSUCHIYA
Keyword(s):  
Rat Kidney ◽  
Aminoglycoside Antibiotics ◽  
Kidney Cortex ◽  
Rat Kidney Cortex

scholarly journals HISTOCHEMICAL DEMONSTRATION OF N-ACETYL-β-GLUCOSAMINIDASE EMPLOYING NAPHTHOL AS-BI N-ACETYL-β-GLUCOSAMINIDE AS SUBSTRATE

1965 ◽  
Vol 13 (5) ◽  
pp. 355-360 ◽  
Author(s):  
MASANDO HAYASHI
Keyword(s):  
Enzyme Activity ◽  
Lysosomal Enzymes ◽  
Rat Kidney ◽  
Histochemical Demonstration ◽  
Frozen Sections ◽  
Aqueous Methanol ◽  
Hepatic Cells ◽  
Cytological Localization ◽  
Convoluted Tubules ◽  
Methanolic Ammonia

The N-acetyl-β-glucosaminide of naphthol AS-BI (7-bromo-3-hydroxy-2-naphth- o-anisidide) was obtained by reacting the anisidide with acetochloroglucosamine in aqueous alkaline acetone. After removal of O-acetyl groups with methanolic ammonia and recrystallization from aqueous methanol, the compound may be used as a substrate for N-acetyl-β-glucosaminidase. Formol-calcium fixed frozen sections were incubated with 0.5 mM substrate in the presence of hexazonium pararosanilin or fast garnet GBC at pH 5.2. Staining appeared in the cytoplasm mostly as discrete granules at the presumed site of enzyme activity and was inhibited in the presence of N-acetylglucosaminolactone. In the rat kidney a number of spherical granules which stained for the enzyme were observed in the proximal portion of the proximal convoluted tubules. In the liver staining was recognized as small granules localized at the pericanalicular cytoplasm of parenchymal hepatic cells. The general cytological localization in these tissues was quite similar to that of other lysosomal enzymes. The limitations of the technique were briefly discussed.

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