A circadian pacemaker, thought to be within the suprachiasmatic nucleus (SCN) of the hypothalamus, begins to function before birth in rodents. Prenatal entrainment of the pacemaker appears to be mediated by signals regulated by the maternal SCN; ablation of the mother's SCN during gestation disrupts the normal phase of the pups' rhythms. The present paper presents an experimental approach for identifying candidate entraining signals and for testing when they are effective during development. The candidate signal examined in these experiments was the pineal gland hormone, melatonin. Female golden hamsters (Mesocricetus auratus) received SCN lesions on day 7 of gestation. During the last week of gestation, they were given two daily subcutaneous injections of oil 12 h apart. One of the injections each day contained melatonin (10, 50, or 100 micrograms). The phases of the pups' activity rhythms were measured at weaning and were found to be related to the timing of the daily injection that contained melatonin, demonstrating that the melatonin directly or indirectly set the phase of the pups' rhythms. Injections given over 4 days of gestation were found to be as effective as injections given over 7 days. Although a physiological role for melatonin as an entraining signal has not been demonstrated, the results show that exogenous, prenatal treatment can predictably set the phase of the offsprings' circadian rhythms.

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Cell determination in vertebrates requires integration of many events, although the mechanisms controlling the different stages in this process are poorly understood. While studies of lens determination have helped define some of these stages, we know very little about the intermediate steps involved in the commitment of ectoderm to lens formation. Lens determination begins during gastrulation when ectoderm is briefly competent to respond to lens-inducing signals and progresses to a point, at the neural tube stage, when the presumptive lens ectoderm is specified. Between these two stages important regulatory genes are activated in the presumptive lens ectoderm, including the transcription factor Pax-6, and transplantation experiments presented here suggest that the presumptive lens ectoderm is becoming "biased" toward lens formation. We show that competent ectoderm from Xenopus laevis embryos forms well-differentiated lenses in most cases when transplanted to the presumptive lens area of neural plate stage hosts, where the lens-inductive environment is strong. When placed into later, neural tube stage hosts, optimally competent ectoderm does form small lenses in about half the cases, but the overall response is much weaker. Even in this weakly inducing environment, however, lens ectoderm that is part way through the inductive process (at the neural plate stage) is shown to have a lens-forming bias, since it forms well differentiated lenses in nearly all cases. While we find that ectoderm surrounding the lens-forming area at neural plate stages does not have a lens-forming bias, non-lens head ectoderm at the neural tube stage does, suggesting that a large region of head ectoderm is biased during neurulation. Using Rana palustris embryos, a species used in the earliest lens induction studies, we were also able to show that the optic vesicle can induce lenses in non-lens head ectoderm at neural tube stages. These results lead us to refine the definition of lens cell determination and provide a context that should allow clarification of determination mechanisms.