SLO3 KO MOUSE: A TOOL TO REVEAL VOLTAGE-DEPENDENT PROCESSES IN SPERM FERTILITY

SLO3 KO 小鼠：揭示精子受精过程中电压依赖性过程的工具

• 批准号: 8469876
• 负责人: Celia M Santi
• 金额: $ 32.46万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-02 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8469876
• 关键词: A Mouse; Accounting; Acrosome Reaction; Affect; Bicarbonates; Bovine Serum Albumin; Cell membrane; Cellular biology; Cholesterol; Clinical; Complement; Contraceptive Agents; Contraceptive methods; Cyclic AMP-Dependent Protein Kinases; Defect; Elements; Environment; Event; Failure; Female; Fertility; Fertilization; Fertilization in Vitro; Genes; Genetic Variation; Genital system; Germ Cells; Grant; Individual; Infertility; Ion Channel; Ionophores; Knock-out; Knockout Mice; Knowledge; Life; Mammals; Membrane; Membrane Potentials; Molecular; Pharmaceutical Preparations; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositols; Physiological; Physiology; Potassium Channel; Procedures; Process; Property; Proteins; Regulation; Role; Series; Signal Pathway; Sperm Capacitation; Sperm Motility; System; Techniques; Testing; Transgenic Organisms; Tyrosine Phosphorylation; Work; Zona Pellucida; cell motility; design; electrical property; improved; in vivo; male; research study; sperm cell; tool; voltage; zygote

DESCRIPTION (provided by applicant): Sperm capacitation is a poorly understood series of molecular processes that occur in vivo in the female genital tract, and is absolutely essential to a sperm's ability to fertilize an egg. A key component of capacitation is the hyperpolarization of the sperm plasma membrane, but the molecular mechanism underlying hyperpolarization was unknown. We previously showed by using a SLO3 knock-out (SLO3 -/-) mouse that the SLO3 potassium channel is the main ion channel responsible for the hyperpolarization that occurs during sperm capacitation and that the SLO3 -/- is male infertile. We also demonstrated that sperm from SLO3 -/- mice have major deficits in sperm motility as well as in the Acrosome Reaction (AR). Since SLO3 channels are the major determinant of changes in membrane potential (Em) that occur during capacitation, it should now be possible to reveal how the components of the capacitating media trigger the activation of SLO3 channels and how SLO3 channel activation affects other signaling pathways that are active during capacitation and the AR. Since SLO3 -/- sperm have impaired motility and AR, and both events require increases in [Ca2+]i, we will investigate how SLO3 channel activation and subsequent changes in Em affect the [Ca2+]i changes that occur during capacitation and the AR. The overall objective of this proposal is to reveal the role of sperm membrane potential (Em) changes in capacitation and the AR. The SLO3 -/- mouse now represents an invaluable tool which permits us to gain greater experimental control over sperm Em and will reveal the importance of Em in each element of sperm physiology examined. The fact that SLO3 channels are essential to fertilization also suggests the possibility that genetic variation within this gene accounts for differences in fertility among male individuals. SLO3 channels also represent an ideal target for designing and testing blockers specific for this essential channel that can be used as contraceptive drugs. Finally, knowledge of the electrical properties of sperm and the essential aspects of the external ionic environment may have important implications for improving the efficiency of clinical IVF procedures.

描述（由申请人提供）：精子获能是女性生殖道体内发生的一系列分子过程，人们对此知之甚少，并且对于精子使卵子受精的能力至关重要。获能的一个关键组成部分是精子质膜的超极化，但超极化背后的分子机制尚不清楚。我们之前通过使用 SLO3 敲除 (SLO3 -/-) 小鼠表明，SLO3 钾通道是导致精子获能期间发生的超极化的主要离子通道，并且 SLO3 -/- 是雄性不育的。我们还证明，SLO3 -/- 小鼠的精子在精子活力和顶体反应（AR）方面存在严重缺陷。由于 SLO3 通道是获能期间发生的膜电位 (Em) 变化的主要决定因素，因此现在应该可以揭示获能介质的成分如何触发 SLO3 通道的激活以及 SLO3 通道激活如何影响其他信号通路在获能和 AR 期间处于活动状态。由于 SLO3 -/- 精子的活力和 AR 受损，并且这两种事件都需要 [Ca2+]i 增加，因此我们将研究 SLO3 通道激活和 Em 的后续变化如何影响获能和 AR 期间发生的 [Ca2+]i 变化。该提案的总体目标是揭示精子膜电位 (Em) 变化在获能和 AR 中的作用。 SLO3 -/- 小鼠现在代表了一种宝贵的工具，它使我们能够对精子 Em 进行更好的实验控制，并将揭示 Em 在所检查的精子生理学每个要素中的重要性。 SLO3 通道对于受精至关重要这一事实也表明该基因内的遗传变异可能解释了男性个体之间生育力的差异。 SLO3 通道还代表了设计和测试针对这一重要通道（可用作避孕药物）的阻断剂的理想靶标。最后，了解精子的电特性和外部离子环境的基本方面可能对提高临床 IVF 程序的效率具有重要意义。