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We report data demonstrating that zebrafish embryos irradiated by alpha particles can release a stress signal into the water, which can be communicated to the unirradiated zebrafish embryos sharing the same water medium and thereby inducing a radioadaptive response in these unirradiated zebrafish embryos. The effects of radiation on the whole embryos were studied through quantification of apoptotic signals at 24 h post fertilization through staining with the vital dye acridine orange, followed by counting the stained cells under a microscope. In these experiments, dechorionated embryos were irradiated and then partnered with two other groups of unirradiated embryos, namely the bystander group (no more further treatments) and adaptive group (subjected to a further challenging dose) of embryos. The adaptive group of embryos were then separately further irradiated with a challenging dose. The results show that the number of apoptotic signals for the adaptive group is smaller than that for the corresponding control group, while that for the bystander group is larger than that for the corresponding control group. These suggest that the stress communicated in vivo between the irradiated zebrafish embryos and those unirradiated embryos sharing the same medium will induce radioadaptive response in the unirradiated embryos.

In the studies reported here, a high-linear-energy-transfer (high-LET)-radiation dose was used to induce adaptive response in zebrafish embryos in vivo. Microbeam protons were used to provide the priming dose and X-ray photons were employed to provide the challenging dose. The microbeam irradiation system (Single-Particle Irradiation System to Cell, acronym as SPICE) at the National Institute of Radiological Sciences (NIRS), Japan, was employed to control and accurately quantify the number of protons at very low doses, viz., about 100 µGy. The embryos were dechorionated at 4 h post fertilization (hpf) and irradiated at 5 hpf by microbeam protons. For each embryo, ten irradiation points were arbitrarily chosen without overlapping with one another. To each irradiation point, 5, 10 or 20 protons each with an energy of 3.4 MeV were delivered. The embryos were returned back to the incubator until 10 hpf to further receive the challenging exposure, which was achieved using 2 Gy of X-ray irradiation, and then again returned to the incubator until 24 hpf for analyses. The levels of apoptosis in zebrafish embryos at 25 hpf were quantified through terminal dUTP transferase-mediated nick end-labeling (TUNEL) assay, with the apoptotic signals captured by a confocal microscope. The results revealed that 5 to 20 protons delivered at 10 points each on the embryos, or equivalently 110 to 430 µGy, could induce radioadaptive response in the zebrafish embryos in vivo.

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