title: Mating rhythms of Drosophila: rescue of tim01 mutants by D. ananassae timeless.
abstract: BACKGROUND
  It is reported that the circadian rhythms of female mating activity differ among Drosophila species and are controlled by an endogenous circadian clock. Here, we found that the mating rhythm of D. ananassae differed from that of D. melanogaster. Moreover, to evaluate the effect of clock gene products on mating activities, we examined the mating activity of D. melanogaster timeless (tim01) transgenic fly harboring heat-shock promotor driven-D. ananassae timeless (tim) gene (hs-AT tim01).
  
  
  METHODS
  Flies were maintained under light/dark (LD) cycles for several days and then they were transferred to constant dark (DD) conditions at 25 degrees C. Transformant flies were heat-shocked for 30 min (PZT 10.5-11.0 or PZT 22.5-23.0; PZT means Projected Zeitgeber Time) at 37 degrees C every day. Daily expressions of D. ananassae TIMELESS (TIM) protein in transgenic flies were measured by western blotting. To examine whether the timing of D. ananassae TIM protein induction by heat shock can change the patterns of the behavior activities of D. melanogaster tim01 flies, we measured locomotor and mating activity rhythms under DD at 25 degrees C +/- 0.5 degrees C except when heat shock was applied.
  
  
  RESULTS
  Heat shock applied at PZT 10.5-11.0 and at PZT 22.5-23.0 induced high TIM levels during subjective night and day, respectively, in hs-AT tim01 flies. The locomotor rhythm of these flies was changed from diurnal to nocturnal by the timing of D. ananassae TIM induction. However, the mating rhythm of these flies could not be entrained by the timing of D. ananassae TIM induction.
  
  
  CONCLUSIONS
  The pattern of mating activity rhythms of D. ananassae and of D. melanogaster differed. The mating activity rhythms of D. melanogaster tim01 flies harboring hs-AT tim appeared after heat-shock but the pattern and phase differed from those of wild-type D. ananassae and D. melanogaster. Moreover, the mating rhythm of these flies could not be entrained by the timing of D. ananassae TIM induction although the locomotor rhythm of hs-AT tim01 was changed from diurnal to nocturnal according to the timing of D. ananassae TIM induction. These data suggest that species-specific mating activities require output pathways different from those responsible for locomotor rhythms.
authors.affiliation: Clock Cell Biology, National Institute of Advanced Industrial Science and Technology, Institute of Molecular and Cell Biology, Tsukuba 305-8566, Japan. n.ishida@aist.go.jp
authors.forename: Izumi
authors.initials: I
authors.lastname: Nishinokubi
authors.forename: Masami
authors.initials: M
authors.lastname: Shimoda
authors.forename: Norio
authors.initials: N
authors.lastname: Ishida