Effect of monensin on the neuronal ultrastructure and endocytic pathway of macromolecules in cultured brain neurons.
1. The endocytic pathway of horseradish peroxidase (HRP) was investigated in the perikarya of cultured neurons by electron microscopy and enzyme cytochemistry. The tracer was observed in endocytic pits and vesicles, endosomes, multivesicular bodies, and lysosomes. It took approximate 15 min for the transfer of HRP from the exterior of the cell to the lysosomes. 2. Monensin induced distension of the Golgi apparatus and formation of intracellular vacuoles. When neurons were incubated with both monensin and HRP for 30 to 120 min, the number of HRP-labeled endosomes was greater than that in the monensin-free group, whereas the reverse was seen for HRP-positive lysosomes. The formation of HRP-positive lysosomes in monensin-treated cells was blocked by 47 to 79%. 3. These results indicate that the intracellular transport of the endocytosed macromolecule is pH dependent. It is also possible that the export of lysosomal enzymes is inhibited by monensin, resulting in an accumulation of the endosomes and a reduction of the lysosomes.
Yin
H S
HS
Department of Anatomy, College of Medicine, National Taiwan University, Taipei, Republic of China.
Yang
M F
MF
D000818:Animals; D001692:Biological Transport; D001921:Brain / Q000166:cytology*; D002478:Cells, Cultured; D002642:Chick Embryo; D004705:Endocytosis / Q000187:drug effects*; D006735:Horseradish Peroxidase / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008247:Lysosomes / Q000378:metabolism; D008854:Microscopy, Electron; D008985:Monensin / Q000494:pharmacology*; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D014617:Vacuoles / Q000378:metabolism
D008985:Monensin; D006735:Horseradish Peroxidase