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Biochem Biophys Res Commun. 1994 Jul 15;202(1):120-7.

Involvement of Ca2+ in the formation of high molecular weight DNA fragments in thymocyte apoptosis.

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Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.


Internucleosomal DNA fragmentation (DNA laddering) and formation of apoptotic bodies have long been considered characteristic features of apoptosis. However, recent work has shown that formation of high molecular weight DNA fragments precedes internucleosomal cleavage and may involve mechanisms that differ from those responsible for DNA laddering. Here, we show that glucocorticoid treatment of human thymocytes stimulated the formation of high molecular weight DNA fragments by Ca(2+)- and endonuclease-mediated mechanisms. Either the removal of Ca2+ from the medium or pretreatment of the cells with the intracellular Ca2+ chelator, BAPTA-AM, prevented the formation of large DNA fragments. Further, treatment of the thymocytes with the microsomal Ca(2+)-ATPase inhibitor, thapsigargin, which caused a sustained increase in intracellular Ca2+ concentration, was in itself sufficient to activate high molecular weight DNA fragmentation. Our results show that Ca(2+)-dependent mechanisms promote the multistep chromatin cleavage in human thymocyte apoptosis.

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