Cellular taurine release triggered by stimulation of the Fas(CD95) receptor in Jurkat lymphocytes.
One of the hallmarks of apoptosis is cell shrinkage which appears to be important for cell death. The mechanisms mediating cell volume decrease have, however, not been addressed. Mechanisms employed by swollen cells to decrease their cell volume include activation of ion transport pathways, such as ion channels and KCl cotransport, and release of cellular osmolytes, such as taurine, sorbitol, betaine and inositol. The present study has been performed to test for release of taurine. To this end Jurkat human T-lymphocytes were loaded with [3H]taurine and apoptotic cell death induced by triggering the Fas(CD95) receptor with monoclonal crosslinking antibody. Triggering the Fas(CD95) receptor led to a release of 60+/-5% of cellular taurine within 90 min. The release did not occur prior to 45 min. The release coincided with cell shrinkage as evidenced from forward scatter in FACS analysis and preceeded DNA fragmentation according to propidium iodide staining. The delay of taurine release was not influenced by exchange of medium and thus was not due to extracellular accumulation of a stimulator. The Fas(CD95)-induced taurine release, cell shrinkage and DNA fragmentation were blunted by lowering of ambient temperature to 23 degreesC. Following pretreatment of cells with Fas(CD95) antibody at 23 degreesC rewarming led to rapid taurine release, cell shrinkage and DNA fragmentation, indicating that the temperature-sensitive step is distal to the mechanisms accounting for the delay. Osmotic cell swelling led to an immediate release of taurine. In conclusion, Fas(CD95) triggering leads to delayed taurine release through a temperature-sensitive mechanism.
Lang
F
F
Physiologisches Institut, der Universität Tübingen, Gmelinstrasse 5, D-72076 Tübingen, Germany.
Madlung
J
J
Uhlemann
A C
AC
Risler
T
T
Gulbins
E
E
D000911:Antibodies, Monoclonal / Q000276:immunology; D048429:Cell Size; D004396:Coloring Agents; D053938:DNA Fragmentation; D053222:Fas Ligand Protein; D006801:Humans; D019169:Jurkat Cells; D008562:Membrane Glycoproteins / Q000276:immunology / Q000378:metabolism*; D009995:Osmosis; D011419:Propidium; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism*; D013654:Taurine / Q000378:metabolism*; D013696:Temperature; D019014:fas Receptor / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D004396:Coloring Agents; C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D019014:fas Receptor; D013654:Taurine; D011419:Propidium