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Fertilization and Development of Quail Oocytes after Intracytoplasmic Sperm Injection¹

Laboratory of Animal Physiology,³ Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Aichi 464-8601, Japan Faculty of Agriculture,⁴ Shinshiu University, Minamiminowa 399-4598, Japan

The present study was conducted to establish the intracytoplasmic sperm injection (ICSI) method for in vitro fertilization and development in quail. The efficiency of fertilization of oocytes was compared 1) between spontaneous and premature ovulation and 2) among testicular round spermatids, elongated spermatids, and immature and mature spermatozoa. The oocytes were injected with a single spermatozoon or spermatid and cultured for 24 h. Cell division was histologically observed with hematoxylin-eosin (HE) and a nucleus-specific fluorescent dye (DAPI). Five of 30 (16.6%) and 4 of 30 (13.3%) oocytes injected with mature sperm were fertilized in the spontaneous and induced ovulation group, respectively. Those embryos showed development at stages II–VII. Half the number (three of six) of the oocytes injected with testicular spermatozoa were fertilized and developed to stages IV–VII, and two of five oocytes injected with elongated spermatids were fertilized and developed to stage VI. All ooocytes injected with round spermatids were unfertilized. The results demonstrate that intracytoplasmic injection of a single sperm into quail oocyte can activate the oocyte and lead to fertilization. Oocytes prematurely ovulated are capable of fertilizing with mature sperm as are those spontaneously ovulated. In addition, the results suggest that the testicular round spermatids may not possess sufficient oocyte-activating potency but that the elongated spermatids and immature spermatozoa are competent to participate in fertilization and early embryonic development in quail.

¹ This research was supported by grant-in-aid from the Japan Society for the Promotion of Science, Postdoctoral Fellowship for Foreign Researchers awarded to A.H. (P 01319), and from the Japan Society for the Promotion of Science, Basic Research (C2) to K.S. (No. 13660286). A.H. on leave from Department of Animal Physiology, Agricultural University of Cracow, Al. Mickiewicza 24/28, 30-059 Krakow, Poland

² Correspondence: Kiyoshi Shimada, Laboratory of Animal Physiology, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Aichi 464-8601, Japan. FAX: 81 52 789 4065; kshimada{at}agr.nagoya-u.ac.jp

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				S. Takagi, T. Ono, A. Tsukada, Y. Atsumi, S. Mizushima, N. Saito, and K. Shimada Fertilization and Blastoderm Development of Quail Oocytes After Intracytoplasmic Injection of Chicken Sperm Bearing the W Chromosome Poult. Sci., May 1, 2007; 86(5): 937 - 943. [Abstract] [Full Text] [PDF]

				S. Takagi, A. Tsukada, N. Saito, and K. Shimada Fertilizing Ability of Chicken Sperm Bearing the W Chromosome Poult. Sci., April 1, 2007; 86(4): 731 - 738. [Abstract] [Full Text] [PDF]