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By In Vivo, Naoufal Zamzami, Maria Castedo, Carole Zanin, Patrice X. Petit and Guido Kroemer

Abstract

In a number of experimental systems in which lymphocyte depletion was induced by apoptosisinducing manipulations, no apoptotic morphology and ladder-type DNA fragmentation were detected among freshly isolated peripheral lymphocytes ex vivo. Here we report that one alteration that can be detected among splenocytes stimulated with lymphocyte-depleting doses of dexamethasone (DEX) in vivo is a reduced uptake of 3,3'dihexyloxacarbocyanine iodide (DiOC6[3]), a fluorochrome which incorporates into cells dependent upon their mitochondrial transmembrane potential (A~m). In contrast, ex vivo isolated splenocytes still lacked established signs of programmed cell death (PCD): DNA degradation into high or low molecular weight fragments, ultrastructural changes of chromatin arrangement and endoplasmatic reticulum, loss in viability, or accumulation of intracellular peroxides. Moreover, no changes in cell membrane potential could be detected. A reduced A~m has been observed in response to different agents inducing lymphoid cell depletion in vivo (superantigen and glucocorticoids [GC]), in mature T and B lymphocytes, as well as their precursors. DEX treatment in vivo, followed by cytofluorometric purification of viable A~m l ~ splenic T cells ex vivo, revealed that this fraction of cells is irreversibly committed to undergoing DNA fragmentation. Immediately after purification neithe

Year: 1995
OAI identifier: oai:CiteSeerX.psu:10.1.1.321.9288
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