Evolution of DNA strand-breaks in cultured spermatocytes: the Comet Assay reveals differences in normal and gamma-irradiated germ cells.
In reproductive toxicity assessment, in vitro systems can be used to determine mechanisms of action of toxicants. However, they generally investigate the immediate effects of toxicants, on isolated germ cells or spermatozoa. We report here the usefulness of in vitro cultures of rat spermatocytes and Sertoli cells, in conjunction with the Comet Assay to analyze the evolution of DNA strand-breaks and thus to determine DNA damage in germ cells. We compared cultures of normal and gamma-irradiated germ cells. In non-irradiated spermatocytes, the Comet Assay revealed the presence of DNA strand-breaks, which numbers decreased with the duration of the culture, suggesting the involvement of DNA repair mechanisms related to the meiotic recombination. In irradiated cells, the evolution of DNA strand-breaks was strongly modified. Thus our model is able to detect genotoxic lesions and/or DNA repair impairment in cultured spermatocytes. We propose this model as an in vitro tool for the study of genotoxic injuries on spermatocytes.
Perrin
J
J
IFR PMSE 112 (Pôle Méditerranéen des Sciences de l'Environnement), Facultés de Médecine et de Pharmacie, Université de la Méditerranée, 27 Boulevard Jean Moulin, 13385 Marseille Cedex 05, France. jeanne.perrin@ap-hm.fr
Lussato
D
D
De Méo
M
M
Durand
P
P
Grillo
J-M
JM
Guichaoua
M-R
MR
Botta
A
A
Bergé-Lefranc
J-L
JL
D000818:Animals; D000963:Antimetabolites; D001973:Bromodeoxyuridine; D020169:Caspases / Q000378:metabolism; D002470:Cell Survival / Q000528:radiation effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D020552:Comet Assay; D004249:DNA Damage / Q000528:radiation effects*; D005720:Gamma Rays; D005854:Germ Cells / Q000528:radiation effects* / Q000648:ultrastructure; D007700:Kinetics; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D012708:Sertoli Cells / Q000528:radiation effects / Q000648:ultrastructure; D013090:Spermatocytes / Q000528:radiation effects* / Q000648:ultrastructure
D000963:Antimetabolites; D020169:Caspases; D001973:Bromodeoxyuridine