Signal transduction mechanisms for the regulation of sperm motility

调节精子活力的信号转导机制

• 批准号: 06660360
• 负责人: ASHIZAWA Koji
• 金额: $ 1.28万
• 依托单位: Miyazaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06660360/
• 关键词: Spermatozoa; Motility; Calcium; Phosphorylation; カルモデュリン

Uniquely, fowl spermatozoa display the phenomenon of reversible, temperature-dependent motility inhibition : in simple salt solutions in vitro, they are immotile at the avian body temperature of 40-41ﾟC,but regain motility on reduction of the temperature to 30ﾟC.However, the intracellular molecular cascades involved in the regulation of motility remain to be determined.Intracellular free Ca^<2+> seems to be essential for the maintenance of motility, since the motility of intact spermatozoa at 40ﾟC can be restored by the addition of Ca^<2+>. Furthermore, the motility of intact spermatozoa loaded with an intracellular Ca^<2+> chelator, BAPTA/AM is negligible at 30ﾟC,but can be instantly restored by the subsequent addition of Ca^<2+>. However, even in the presence of Ca^<2+>, the addition of calmodulin antagonists such as W-7 and trifluoperazine inhibited sperm motility at 30ﾟC and 40ﾟCThe activation of protein phosphatase type 1 (PP1), present in the fowl sperm axoneme, may be involved i … More n the inhibition of fowl sperm motility at 40ﾟC,since, in addition to calyculin A and okadaic acid, specific inhibitors of PP1, inhibitors 1 and 2, also stimulated the motility of demembranated spermatozoa at 40ﾟC.Furthermore, the involvement of myosin light chain kinase (MLCK), or a MLCK-like protein, has been proposed to be a candidate of Ca^<2+>/calmodulin dependent protein kinase, since the motility of both intact and demembranated fowl spermatozoa at 30ﾟC decreased markedly following the addition of ML-9, a specific inhibitor of MLCK,but not following the addition of H-8, a specific inhibitor or cAMP-dependent protein kinase (PKA). A marked difference in phosphorylation was observed in a 30 kDa protein. This protein was strongly phosphorylated after the addition of Kemptide, a PKA substrate peptide as well as control samples but only slightly phosphorylated in the presence of a myosin light chain kinase (MLCK) substrate peptide.These results suggest that phosphorylation by Ca^<2+>/calmodulin-dependent protein kinases, rather tha PKA,is likely to be a regulatory step in the maintenance of fowl sperm motility, and that phosphorylation of a 30 kDa protein by MLCK or a MLCK-like proteinis may be involved in the regulation of flagellar movement at 30ﾟC. Less

独特的是，家禽精子显示可逆的、温度依赖性运动抑制现象：在体外的简单盐溶液中，它们在40-41 ℃的禽体温下不动，但在温度降低到30 ℃时恢复运动。然而，参与运动调节的细胞内分子级联仍有待确定。细胞内游离Ca^<2+> Ca ^<2 +>似乎对维持精子的活动力是必不可少的，因为在40 ℃时，完整精子的活动力可以通过加入Ca^<2+>而恢复。此外，在30 ℃时，装有细胞内Ca^<2+>螯合剂BAPTA/AM的完整精子的运动力可以忽略不计，但随后加入Ca^<2+>可以立即恢复。然而，即使在Ca^<2+>存在的情况下，加入钙调素拮抗剂如W-7和三氟拉嗪在30 ℃和40 ℃下也能抑制精子运动。 ...更多信息 在40 ℃下抑制家禽精子运动，因为除了calyculin A和冈田酸外，PP 1的特异性抑制剂，抑制剂1和2，也在40 ℃下刺激脱膜精子的运动。此外，肌球蛋白轻链激酶（MLCK）或MLCK样蛋白的参与被认为是Ca^2+/钙调蛋白依赖性蛋白激酶的候选者，因为在30 ℃下，完整和去膜的家禽精子的运动性在加入MLCK的特异性抑制剂ML-9后显著降低，但在加入H-8（cAMP依赖性蛋白激酶（PKA）的特异性抑制剂）后不显著降低。在30 kDa蛋白质中观察到磷酸化的显著差异。在加入PKA底物肽Kemptide和对照样品后，该蛋白被强烈磷酸化，但在肌球蛋白轻链激酶（MLCK）底物肽存在下仅轻微磷酸化。这些结果表明，Ca^2+/钙调蛋白依赖性蛋白激酶的磷酸化，而不是PKA，可能是维持家禽精子活力的调节步骤，MLCK或MLCK样蛋白对30 kDa蛋白的磷酸化可能参与30 ℃下鞭毛运动的调节。少

项目成果

Ashizawa, K.et al.: "Presence of protein phosphatase type 1 and its involvement in temperature-dependent flagellar movement of fowl spermatozoa." FEBS Lett.350. 130-134 (1994)

Ashizawa, K.等人：“1 型蛋白磷酸酶的存在及其参与鸡精子温度依赖性鞭毛运动。”

Ashizawa, K.et al.: "Regulatory mechanisms of fowl sperm motility : possible role of endogenous myosin light chain kinase-like protein." J.Reprod.Fertil.104. 141-148 (1995)

Ashizawa, K.等人：“家禽精子活力的调节机制：内源性肌球蛋白轻链激酶样蛋白的可能作用。”

Ashizawa, K.et al.: "Stimulation of motility and respiration of intact fowl spermatozoa by calyculin A,a specific inhibitor of protein phosphatase-1 and -2A,via a Ca^<2+> dependent manner." J.Reprod.Fertil.105. 109-114 (1995)

Ashizawa, K.等人：“花萼蛋白 A（蛋白磷酸酶-1 和 -2A 的特异性抑制剂）通过 Ca^2 依赖性方式刺激完整家禽精子的运动和呼吸。”

Ashizawa,K. et al: "Regilatory mechanisms of fowl sperm motility : possible role of endogenous myosin light chain kinase-like protein." J. Reprod. Fertil.104. 141-148 (1995)

芦泽，K.

Ashizawa,K. et al: "Stimulatory of motility and respiration of intact fowl spermatozoa by calyculin A, a specific inhibitor of protein phosphatase-1 ans -2A, via a Ca^<2++> dependent" J. Reprod. Fertil.105. 109-114 (1995)

芦泽，K.