title: The birth of quail chicks after intracytoplasmic sperm injection.
abstract: Intracytoplasmic sperm injection (ICSI) has been successfully used to produce offspring in several mammalian species including humans. However, ICSI has not been successful in birds because of the size of the egg and difficulty in mimicking the physiological polyspermy that takes place during normal fertilization. Microsurgical injection of 20 or more spermatozoa into an egg is detrimental to its survival. Here, we report that injection of a single spermatozoon with a small volume of sperm extract (SE) or its components led to the development and birth of healthy quail chicks. SE contains three factors - phospholipase Cζ (PLCZ), aconitate hydratase (AH) and citrate synthase (CS) - all of which are essential for full egg activation and subsequent embryonic development. PLCZ induces an immediate, transient Ca(2+) rise required for the resumption of meiosis. AH and CS are required for long-lasting, spiral-like Ca(2+) oscillations within the activated egg, which are essential for cell cycle progression in early embryos. We also found that co-injection of cRNAs encoding PLCZ, AH and CS support the full development of ICSI-generated zygotes without the use of SE. These findings will aid our understanding of the mechanism of avian fertilization and embryo development, as well as assisting in the manipulation of the avian genome and the production of transgenic and cloned birds.
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan.
authors.forename: Shusei
authors.initials: S
authors.lastname: Mizushima
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan.
authors.forename: Gen
authors.initials: G
authors.lastname: Hiyama
authors.affiliation: Shimoda Marine Research Center, University of Tsukuba, 5-10-1 Shimoda, Shizuoka 415-0025, Japan.
authors.forename: Kogiku
authors.initials: K
authors.lastname: Shiba
authors.affiliation: Shimoda Marine Research Center, University of Tsukuba, 5-10-1 Shimoda, Shizuoka 415-0025, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Inaba
authors.affiliation: Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan.
authors.forename: Hideo
authors.initials: H
authors.lastname: Dohra
authors.affiliation: Department of Food Production Science, Faculty of Agriculture, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano 399-4598, Japan.
authors.forename: Tamao
authors.initials: T
authors.lastname: Ono
authors.affiliation: WCU Major in Biomodulation, Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Shimada
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan atsasan@ipc.shizuoka.ac.jp.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Sasanami
mesh_terms: D000154:Aconitate Hydratase / Q000032:analysis; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002853:Chromatography, Liquid; D002950:Citrate (si)-Synthase / Q000032:analysis; D005306:Fertilization / Q000502:physiology*; D015151:Immunoblotting; D008297:Male; D008856:Microscopy, Fluorescence; D010063:Ovum / Q000378:metabolism; D054801:Phosphoinositide Phospholipase C / Q000032:analysis; D011784:Quail / Q000502:physiology*; D020554:Sperm Injections, Intracytoplasmic / Q000379:methods / Q000662:veterinary*; D013094:Spermatozoa / Q000737:chemistry*; D053719:Tandem Mass Spectrometry; D016896:Treatment Outcome
chemical_list: D002950:Citrate (si)-Synthase; D054801:Phosphoinositide Phospholipase C; D000154:Aconitate Hydratase; D002118:Calcium
keywords: Aconitate hydratase; Citrate synthase; Intracytoplasmic sperm injection; Phospholipase Cζ; Physiological polyspermy; Quail