Grant-in-Aid for Scientific Research (B).
|Section||Joint Research .|
|Research Institution||OKAZAKI NATIONAL RESEARCH INSTITUTE|
MOHRI Hideo NATIONAL INSTITUTE FOR BASIC BILOGY, OKAZAKI NATIONAL RESEARCH INSTITUTES, DIRECTOR GENERAL, 基礎生物学研究所, 所長 (70012268)
SAVAGE J.STE ニューオリンズ大学, 生物科学, 副教授
SUAREZ S.S. コーネル大学, 獣医学部, 副教授
YANAGIMACHI ハワイ大学, 医学部, 教授
ISIJIMA Sumio BIOL., LAB., FAC. BIOSCI. & TOKYO INT. TECH, RES. ASSOCIATE, 理学部, 助手 (70193315)
OKUNO Makoto DEPT. BIOL, LAB., FAC. BIOSCI. & BIOTECH, TOKYO SINST. TECH, OKAZAKI NATIONAL RESEARCH INSTITUTES, RES. ASSOCIATE, 大学院・総合文化研究科, 助教授 (40143325)
STEWART・SAVA ニューオリンズ大学, 生物科学, 副教授
岡部 勝 大阪大学, 遺伝情報実験施設, 教授 (30089875)
小川 和男 岡崎国立共同研究機構, 基礎生物学研究所, 助教授 (30132731)
YANAGIMACHI R. DEP. OF ANATOMY & REPROD. IO., UNIV. OF HAWAII, PORFESSOR
SAVAGE J.s. DEP. BIOL., UNIV. NEW ORLEANS, ASSOCIATE PROFESSOR
SUAREZ S.s. DEP. BIOMEDICAL SCI., CORNELL UNIV., PROFESSOR
|Project Fiscal Year
1997 – 1999
Completed(Fiscal Year 1999)
|Budget Amount *help
¥9,400,000 (Direct Cost : ¥9,400,000)
Fiscal Year 1999 : ¥2,100,000 (Direct Cost : ¥2,100,000)
Fiscal Year 1998 : ¥3,200,000 (Direct Cost : ¥3,200,000)
Fiscal Year 1997 : ¥4,100,000 (Direct Cost : ¥4,100,000)
|Keywords||Mammalian perm / hyperactivation / capacitation / flagellar motility / acrosome reaction / sperm-egg interactin / 哺乳類精子 / 超活性化 / 受精能獲得 / 鞭毛運動 / 先体反応 / 卵・精子相互作用 / 透明帯通過|
The main results obtained for these three years are as follows.
(1) Analysis of the records on high speed video system indicated that hamster spermatozoa exhibit an increase in bend of the mid-piece region with an asymmetrical wave form at the beginning of hyperactivation following capacitation. Onset of the acrosome reaction caused a larger bend of the mid-piece region with a symmetrical wave form. Similar results were obtained with monkey spermatozoa, although little change was observed when spermatozoa of transgenic mouse lines, that accumulate mutated green fluorescence protein in the acrosome, were capacitated and acrosome-reacted.
(2) Hyperactivation was induced with procaine in hamster and bull spermatozoa. Lysolecithin also caused simultaneous hyperactivation in guinea pig spermatozoa.
(3) Mammalian spermatozoa show sequential changes in motility, namely resting, weakly active, activated and hyperactivated. Different specific proteins were phosphorylated at each step. Upon hyperactivation a 80kDa protein localized in the main part of sperm flagella was phosphorylated.
(4) CaィイD12+ィエD1 and cAMP affected both amplitude and wave form of sperm flagella. It was found that the redundant nuclear envelope in the base of the flagellum serves as a CaィイD12+ィエD1 store. The CaィイD12+ィエD1appears to be released to the mitochondria and to increase production of ATP to support hyperactivation.
(5) The spermatozoon passing though the zona pellucida exhibited flagella movement with large amplitude and symmetrical wave form. The movement would provide sufficient propulsive force to pass through the zona pellucida.
(6) Mechanism of motility Suppression by glucose was examined. It was found that two of light chains of dynein, motor protein for flagella motility, correspond to Tctex (t-complex) genes.
(7) In addition, fertilization and pregnancy following intracytoplasmic injection of sperm or spermatogenic cells in the mouse, etc., were also examined.