[The microelectrophoresis of the DNA in individual intact and gamma-irradiated thymocytes].
The in vitro gamma-irradiated mouse thymocytes were embedded in low melting agarose at 37 degrees C. After getting at 4 degrees C, the cells were lysed in neutral detergent solution containing proteinase K and ethidium bromide. Microscopic visualization of single lysed and stained cells showed the presence of the central "core" (nuclear matrix) surrounded with "halo" (relaxed nuclear DNA). During electrophoresis (2-5 V/sm, 5 min) this "halo" migrated towards the anode forming a "tail". The use of microdensitometric system provided measuring the size of the tail (L) and quantity of migrated DNA (S) for individual cells as well as obtaining the distribution of these parameters among the cells. The latter may be characteristic of heterogeneity of the cell population. It was shown that L and S increased linearly with the dose irradiation at least between 0.2 and and 5.0 Gy. In irradiated thymocyte (3 Gy) the DNA repair occurred within 10-20 min, but residual DNA damage could be observed even after 60 min of incubation. These damages may initiate the degradation of DNA in irradiated thymocytes that was observed after the repair of DNA.
V A
VA
Tronov
E V
EV
Grin'ko
D R
DR
Beritashvili
I V
IV
Filippovich
D000818:Animals; D004247:DNA / Q000032:analysis / Q000528:radiation effects*; D004249:DNA Damage / Q000528:radiation effects*; D004260:DNA Repair / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D019286:Endopeptidase K; D004996:Ethidium; D005720:Gamma Rays; D007202:Indicators and Reagents; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D012697:Serine Endopeptidases; D013950:Thymus Gland / Q000737:chemistry / Q000528:radiation effects*
D007202:Indicators and Reagents; D004247:DNA; D012697:Serine Endopeptidases; D019286:Endopeptidase K; D004996:Ethidium
Mikroélektroforez DNK individual'nykh intaktnykh i gamma-obluchennykh timotsitov.