Cytochemical examination of the compartments involved in the transcellular transport of horseradish peroxidase in rat hepatocytes.
Horseradish peroxidase (HRP, 10 mg/100 g body weight) was intravenously injected into rats in order to investigate the nature of the compartments involved in the transcellular transport of the protein through hepatocytes into bile. Double cytochemistry for HRP and the marker enzymes for cytoplasmic organelles was used. HRP was shown to be taken up by hepatocytes via vesicles at the sinusoidal surface, some of which were positive for 5'-nucleotidase activity. HRP was then found in the smooth-surfaced vesicles and tubules which were negative in 5'-nucleotidase, glucose 6-phosphatase, thiamine pyrophosphatase and acid phosphatase activity, suggesting that the tubular structures are neither the endoplasmic reticulum, the Golgi apparatus nor lysosomes. Biochemical studies revealed that the lead procedures used for the double cytochemistry did not inhibit the peroxidatic activity of HRP, and conversely that HRP did not interfere with the marker enzyme activity. Such cytochemical observations seemed to be supported by the observation that administration of monensin (3.5 mg/100 g) and chloroquine (5 mg/100 g), which markedly altered the structure of the Golgi apparatus and lysosomes, respectively, slightly altered the biliary excretion of HRP but not to a significant extent.
Mori
M
M
Department of Pathology, Sapporo Medical College, Japan.
Yamaguchi
J
J
Oyamada
M
M
Enomoto
K
K
Kaneko
A
A
D015720:5'-Nucleotidase / Q000378:metabolism; D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D002738:Chloroquine; D005958:Glucosephosphates / Q000378:metabolism; D006651:Histocytochemistry / Q000379:methods; D006735:Horseradish Peroxidase / Q000008:administration & dosage / Q000493:pharmacokinetics*; D007275:Injections, Intravenous; D008099:Liver / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D008854:Microscopy, Electron; D008985:Monensin; D015388:Organelles / Q000201:enzymology / Q000502:physiology* / Q000648:ultrastructure; D051381:Rats; D011919:Rats, Inbred Strains; D013834:Thiamine Pyrophosphatase / Q000378:metabolism
D005958:Glucosephosphates; D002738:Chloroquine; D008985:Monensin; D006735:Horseradish Peroxidase; D000135:Acid Phosphatase; D015720:5'-Nucleotidase; D013834:Thiamine Pyrophosphatase