In the Danio species, interspecific hybridization has been conducted in several combinations. Among them, only the hybrid between a zebrafish (D. rerio) female and a spotted danio (D. nigrofasciatus) male was reported to be fertile. However, beyond these investigations, by means of reproductive biology, gametes of the hybrid have also not been investigated genetically. For this study, we induced a hybrid of the D. rerio female and D. nigrofasciatus male in order to study its developmental capacity, reproductive performance and gametic characteristics. Its hybrid nature was genetically verified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the rhodopsin gene. Almost all the hybrids (36/37) were males, and only one was female. Developing oocytes were observed in the hybrid female, but ovulated eggs have not been obtained thus far. Microscopic observation revealed various head sizes of sperm in the hybrid males. Flow cytometry showed that the hybrid males generated aneuploid sperm with various ploidy levels up to diploidy. In backcrosses between D. rerio females and hybrid males, fertilization rates were significantly lower than the control D. rerio, and most resultant progeny with abnormal appearance exhibited various kinds of aneuploidies ranging from haploidy to triploidy, but only one viable progeny, which survived more than four months, was triploid. This suggested the contribution of fertile diploid sperm of the hybrid male to successful fertilization and development.
Skewed sex ratio has often been observed in hybrids, but the sex was always determined by gonadal phenotypes. Here, we artiˆcially induced reciprocal hybrids between pink salmon and masu salmon and then examined the genetic sex ratio in the hybrids based on the presence or absence of the sex-determining gene sdY. As a result, the sex of 8-month-old hybrids between pink salmon female × masu salmon male and masu salmon female × pink salmon male skewed to female, while 12-month-old hybrids between masu salmon female × pink salmon male were exclusively male. Since the sex ratio between the early embryos and hatching alevin was 1:1, it was consi-
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