TY - JOUR
T1 - A role for calcium in regulating apoptosis in rat thymocytes irradiated in vitro
AU - Story, M. D.
AU - Stephens, L. C.
AU - Tomasovic, S. P.
AU - Meyn, R. E.
N1 - Funding Information:
We thank Mr Gary Lekander and Ms Susan Tsujimoto for technical assistance, and Mrs Lori Verhalen, Mrs Donna Goodman and Miss Sherry Powell for help in the preparation of the manuscript . This investigation was supported by PHS grant number CA-06294 awarded by the National Cancer Institute, DHHS . Supported in part by Grant NIH-NCI CA-16672 awarded by the National Cancer Institute . Animals are maintained in facilities approved by the American Association for Accreditation of Laboratory Animal Care, and in accordance with current United States Department of Agriculture, Department of Health and Human Services, and National Institutes of Health regulations and standards .
PY - 1992
Y1 - 1992
N2 - Thymus-derived lymphocytes undergo death after γirradiation via a pathway termed apoptosis, or programmed cell death. An early step in this pathway is the production of nucleosome-sized fragments of DNA. DNA fragmentation was used as the endpoint in these investigations to examine apoptosis in lymphocytes extracted from the rat thymus and irradiated in vitro. In unirradiated thymocytes the level of DNA fragmentation rose to 15% by the first hour of culture, where it remained approximately constant until the fifth hour. In contrast, thymocytes irradiated with a dose of 2.5 Gy exhibited a large and dramatic increase in DNA fragmentation beginning 2h postirradiation. DNA fragmentation measured 6h after irradiation was detected after as little as 0.25 Gy and reached a maximum of 90% with 10Gy. Metabolic control of DNA fragmentation after irradiation was evidenced by the suppression of DNA fragmentation when thymocytes were incubated with cyclohexamide or actinomycin D. When γirradiated thymocytes were incubated with the Ca2+ chelator EGTA, DNA fragmentation was reduced significantly. BAPTA-AM, a highly specific intracellular Ca2+ chelator, essentially eliminated DNA fragmentation in cells irradiated with 2.5Gy and, unlike EGTA, eliminated the background level of fragmentation in unirradiated samples. Therefore, our data are consistent with the possibility that Ca2+ serves as a second messenger to induce DNA fragmentation in irradiated thymocytes, suggesting a common pathway for cells prompted to enter apoptosis from seemingly dissimilar interval events.
AB - Thymus-derived lymphocytes undergo death after γirradiation via a pathway termed apoptosis, or programmed cell death. An early step in this pathway is the production of nucleosome-sized fragments of DNA. DNA fragmentation was used as the endpoint in these investigations to examine apoptosis in lymphocytes extracted from the rat thymus and irradiated in vitro. In unirradiated thymocytes the level of DNA fragmentation rose to 15% by the first hour of culture, where it remained approximately constant until the fifth hour. In contrast, thymocytes irradiated with a dose of 2.5 Gy exhibited a large and dramatic increase in DNA fragmentation beginning 2h postirradiation. DNA fragmentation measured 6h after irradiation was detected after as little as 0.25 Gy and reached a maximum of 90% with 10Gy. Metabolic control of DNA fragmentation after irradiation was evidenced by the suppression of DNA fragmentation when thymocytes were incubated with cyclohexamide or actinomycin D. When γirradiated thymocytes were incubated with the Ca2+ chelator EGTA, DNA fragmentation was reduced significantly. BAPTA-AM, a highly specific intracellular Ca2+ chelator, essentially eliminated DNA fragmentation in cells irradiated with 2.5Gy and, unlike EGTA, eliminated the background level of fragmentation in unirradiated samples. Therefore, our data are consistent with the possibility that Ca2+ serves as a second messenger to induce DNA fragmentation in irradiated thymocytes, suggesting a common pathway for cells prompted to enter apoptosis from seemingly dissimilar interval events.
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U2 - 10.1080/09553009214550871
DO - 10.1080/09553009214550871
M3 - Article
C2 - 1351912
AN - SCOPUS:0026565365
SN - 0955-3002
VL - 61
SP - 243
EP - 251
JO - International Journal of Radiation Biology
JF - International Journal of Radiation Biology
IS - 2
ER -