Internalization of the inhibitor of the N-acetylgalactosaminyltransferase by chicken embryonic retina cells: reversibility of the inhibitor effects.
Retina cells from 6-day-old chicken embryos were cultured in the presence of an 125I-labeled protein inhibitor of the UDP-N-acetylgalactosamine:GM3,N-acetylgalactosaminyltransferase. The cells were labeled and did not lose the incorporated radioactivity when treated with 0.125% trypsin or 1 M NaCl at 37 degrees C for 1 hr, indicating that the iodinated inhibitor was inside the cells. Immunostaining procedures using an anti-inhibitor antibody were applied to the cells cultured in the presence of the inhibitor after permeabilization of the cells. The inhibitor was found inside the round cells virtually devoid of neurites, but not in flat glial-like cells or in process-bearing neural cells. Also found was an apparent self-recovery effect of the cells for both the anti-neuritogenic effect and the modification of the pattern of labeled gangliosides produced by the inhibitor when the agent was withdrawn from the culture medium after the initial period of 20 hr. This recovery was clearly observed 72 hr after the removal of the inhibitor.
Quiroga
S
S
Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Argentina.
Panzetta
P
P
Caputto
R
R
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002642:Chick Embryo; D005455:Fluorescent Antibody Technique; D005677:G(M1) Ganglioside / Q000378:metabolism; D005700:Galactosyltransferases / Q000037:antagonists & inhibitors*; D005732:Gangliosides / Q000378:metabolism; D007150:Immunohistochemistry; D007457:Iodine Radioisotopes; D017350:N-Acetylgalactosaminyltransferases; D012160:Retina / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*
D005732:Gangliosides; D007457:Iodine Radioisotopes; D005677:G(M1) Ganglioside; D005700:Galactosyltransferases; D017350:N-Acetylgalactosaminyltransferases; C003742:UDPgalactosamine-galactose acetylgalactosaminyltransferase