Calcium entry into keratinocytes induces exocytosis of lysosomes.
During the final steps of epidermal differentiation, extracellular calcium ions enter keratinocytes and induce transglutaminase activity and cornified envelope formation. In other cell types, entry of calcium mediated by ionophores has been reported to induce exocytosis of lysosomes. In this study, we investigated whether lysosomes of keratinocytes might exhibit a similar behaviour. Ionomycin treatment induced cornified envelope formation in keratinocytes, but also morphological changes including plasma membrane blebbing, although no immediate alteration in cell viability could be detected. The activity of the soluble lysosomal enzymes cathepsin C and beta-galactosidase in the culture medium was increased upon ionomycin treatment. Cell leakage did not seem to be responsible for this phenomenon, as suggested by measurements of the cytosolic enzymes adenylate kinase and dipeptidylpeptidase III in the culture medium. Metabolic labelling followed by immunoprecipitation showed that ionomycin induced release of cathepsin D into the culture medium. Simultaneously, lysosome-associated membrane proteins (Lamps) 1 and 2 were detected at the cell surface of ionomycin-treated keratinocytes by biochemical and morphological approaches. These results suggest that upon ionomycin treatment, calcium entry stimulates exocytosis of lysosomes in keratinocytes.
Jans
Ralph
R
Département Histologie-Embryologie, Facultés Universitaires Notre-Dame de la Paix, 5000 Namur, Belgium.
Sartor
Marc
M
Jadot
Michel
M
Poumay
Yves
Y
D000328:Adult; D015703:Antigens, CD / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D020789:Cathepsin C / Q000378:metabolism; D002402:Cathepsin D / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003470:Culture Media / Q000378:metabolism; D005089:Exocytosis / Q000502:physiology*; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology / Q000648:ultrastructure; D051907:Lysosomal Membrane Proteins; D008247:Lysosomes / Q000378:metabolism*; D001616:beta-Galactosidase / Q000378:metabolism
D015703:Antigens, CD; D003470:Culture Media; D007476:Ionophores; D051907:Lysosomal Membrane Proteins; D015759:Ionomycin; D001616:beta-Galactosidase; D020789:Cathepsin C; D002402:Cathepsin D; D002118:Calcium