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Fertilization and Blastoderm Development of Quail Oocytes After Intracytoplasmic Injection of Chicken Sperm Bearing the W Chromosome

S. Takagi^*, T. Ono, A. Tsukada^*, Y. Atsumi^,, S. Mizushima^*, N. Saito^* and K. Shimada^*,1

^* Laboratory of Animal Physiology, Graduate School of Bioagricultural Sciences, Nagoya University, Aichi 464-8601, Japan; and Faculty of Agriculture, Department of Food Production Science, and Department of Bioscience and Food Production Science, Interdisciplinary Graduate School of Science and Technology, Shinshu University, Minamiminowa, Nagano 399-4598, Japan

¹ Corresponding author: kshimada{at}agr.nagoya-u.ac.jp

Our previous study demonstrated that elongated spermatids and sperm carrying the female-specific W-chromosome of the sex-reversed domestic fowl can activate the mouse oocyte, but whether they can fertilize the avian oocyte and lead to a developing zygote remains undetermined. A single sperm isolated from the semen and testis of normal rooster and from a testis of sex-reversed hen was microinjected into a quail oocyte and cultured for 20 to 24 h. Blastoderms were fixed, cleaved, nuclei stained by 4',6'-diamidino-2-phenylin-dole, and developmental stages were assessed. In the normal rooster group, ejaculated and testicular sperm induced blastodermal development in 22.6 and 20% of the quail oocytes, respectively. The developmental stages ranged from IV to VII. In the sex-reversal group, 20% of injected testicular sperm induced blastodermal development. The blastodermal stages varied from stage III to VI. Blastoderms after 4',6'-diamidino-2-phenylindole staining were assayed by PCR to identify the W chromosome of either chicken sperm or quail oocyte. The PCR assay results showed that 2 out of 9 developed blastoderms microinjected with sperm of sex-reversed hen were identified containing the female-specific W chromosome derived from sex-reversed hen. From these results, it is concluded that chicken sperm bearing the W chromosome possess fertilizing ability and can function to stimulate blastoderm development similar to that of normal chicken sperm carrying the Z chromosome.

Key Words: sex reversal • W chromosome • sperm • fertilization • blastodermal development