Quantitative detection of apoptotic thymocytes in low-dose X-irradiated mice by an anti-single-stranded DNA antibody.
The quantitative detection of apoptotic cells in low frequency in the thymus of mice irradiated with X-rays using an anti-single-stranded DNA antibody was explored. The antibody against single-stranded DNA (anti-ssDNA) was obtained with rabbits hyperimmunized with complexes of alkaline-denatured calf thymus DNA. AKR female mice were irradiated with 10 to 100 cGy or 4 Gy X-rays; thereafter, thymus sections were prepared at various times after irradiation. The detection and counting of apoptotic cells in the section were performed after histochemical staining using an anti-ssDNA antibody. The results demonstrate that, although sensitive and quantitative detection of apoptotic cells in irradiated thymus using the anti-ssDNA antibody is possible, the sensitivity is lower compared to that of in situ end-labeling methods, such as TUNEL or ISEL. The antibodies could also be used for rat thymus and spleen. In addition, an increase in positively stained cells by both methods was detected as early as 6 min after the irradiation of mice.
Fujita
K
K
Department of Biology, Toho University School of Medicine, Tokyo, Japan. abfujita@criepi.denken.or.jp
Kawarada
Y
Y
Terada
K
K
Sugiyama
T
T
Ohyama
H
H
Yamada
T
T
D000818:Animals; D017209:Apoptosis; D002417:Cattle; D002452:Cell Count; D004396:Coloring Agents; D004247:DNA / Q000528:radiation effects*; D004249:DNA Damage; D053938:DNA Fragmentation; D004260:DNA Repair; D004277:DNA, Single-Stranded / Q000032:analysis* / Q000276:immunology; D004307:Dose-Response Relationship, Radiation; D004801:Eosine Yellowish-(YS); D005069:Evaluation Studies as Topic; D005260:Female; D005434:Flow Cytometry; D006416:Hematoxylin; D007074:Immunoglobulin G / Q000276:immunology*; D020287:In Situ Nick-End Labeling; D051379:Mice; D008806:Mice, Inbred AKR; D020401:Primed In Situ Labeling; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D013045:Species Specificity; D013194:Staining and Labeling; D013601:T-Lymphocytes / Q000737:chemistry / Q000528:radiation effects*; D013950:Thymus Gland / Q000166:cytology / Q000528:radiation effects*
D004396:Coloring Agents; D004277:DNA, Single-Stranded; D007074:Immunoglobulin G; D004247:DNA; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin