A reliable method for quantitating chromatin fragments by flow cytometry to predict the effect of total body irradiation and hyperthermia on mice.
The frequencies of chromatin fragments, including micronuclei, in murine thymus cells, spleen cells and bone marrow cells have been used as a quantitative indicator of gamma-ray induced chromosome damage and could be used to screen potential radioprotective agents as well. The yield of chromatin fragments induced in mice receiving different dosage levels of total body irradiation alone and in mice also given whole body hyperthermia as a potent radioprotector were assessed by flow cytometric analysis. Our results demonstrated that chromatin fragments induced by irradiation in vivo was clearly dose-dependent and that chromatin fragments could potentially serve as a biological indicator of radiation damage. One hour of whole body hyperthermia at 40 degrees C (+/- 0.2 degree C) given 20 hours before a lethal dosage (900 cGy) of total body irradiation protects 100% of DBA/2 mice from an LD 100/16 irradiation dose (dose of irradiation that killed 100% of the mice in 16 days). This is in good agreement with the percent of chromatin fragments formed in the cells of the protected animals, which showed no significant difference from those observed in the normal mice. The results indicate that whole body hyperthermia protected the thymus and bone marrow from irradiation damage. This study provides further evidence which supports that whole body hyperthermia can act as a potent radioprotector in vivo. Measurement of the frequencies of chromatin fragments by flow cytometry is simple and reliable. The method can be applied to screen radioprotective agents.
Shen
R N
RN
Dept. of Radiation Oncology, Indiana University School of Medicine, Indianapolis 46202-5289.
Crabtree
W N
WN
Wu
B
B
Young
P
P
Sandison
G A
GA
Hornback
N B
NB
Shidnia
H
H
D000818:Animals; D002843:Chromatin / Q000528:radiation effects*; D002869:Chromosome Aberrations; D005260:Female; D005434:Flow Cytometry; D006979:Hyperthermia, Induced; D051379:Mice; D008811:Mice, Inbred DBA; D015162:Micronucleus Tests; D014916:Whole-Body Irradiation / Q000009:adverse effects*
D002843:Chromatin