Differential DNA-repair activity in prespermiogenic cells of various mouse strains

The present report demonstrates differential DNA-repair activity among 14 strains of immature (20 +/- 2 days old) male mice (inbred strains: C57BL/6J, RF/J, Nude homo/nu, RIII/2J, PL/J, AKR/J, Nude hetero/nude, C3H/HeJ, SWR/J, SM/J, ST/J, LP/J, BALB/cJ and random-bred strain: CD-1). The prespermiogenic cells were isolated and enriched by collagenase-trypsin digestion of seminiferous tubules and subsequent 3% albumin-gradient centrifugation. Enriched prespermiogenic cells demonstrated a viability greater than 95% by trypan blue exclusion criteria. For in vitro unscheduled DNA synthesis (UDS) determination, prespermiogenic cells (10(6) cells/ml) were incubated with methyl methanesulfonate (0.4 mM) in the presence of 20 mM hydroxyurea (HU). At 20 mM HU concentration, 90% of S-phase DNA activity in prespermiogenic cells was inhibited and thus, the net UDS activity following MMS exposure was readily determined. MMS-induced UDS activity in the CD-1 mouse strain was both linear up to 4 h of incubation and dose-dependent at 4 h of incubation. The apparent Km for MMS-induced UDS activity in prespermiogenic cells was approx. 1.8 x 10(-4) M. Of the 14 mice strains tested, C57BL/6J and RF/J exhibited the highest DNA-repair activity, while BALB/cJ, LP/J, and ST/J showed the lowest. A maximal difference in UDS activity of 3.5-fold was observed between C57BL/6J and BALB/cJ. Furthermore, a 2.5-fold difference was also noted between RF/J and LP/J mouse strains. Thus, wide variations in DNA-repair activity among 14 mouse strains were clearly demonstrated. Whether genetically select mouse strains with the lowest DNA-repair activity should have greater sensitivity toward environmental mutagens needs to be tested.