Department of Cardiology, Howard Hughes Medical Institute, Manton Center for Orphan Disease, Children's Hospital Boston, Boston, MA 02115, USA.
Proc Natl Acad Sci U S A. 2011 Aug 23;108(34):14342-7. doi: 10.1073/pnas.1111695108. Epub 2011 Aug 10.
Sperm cells acquire hyperactivated motility as they ascend the female reproductive tract, which enables them to overcome barriers and penetrate the cumulus and zona pellucida surrounding the egg. This enhanced motility requires Ca(2+) entry via cation channel of sperm (CatSper) Ca(2+)-selective ion channels in the sperm tail. Ca(2+) entry via CatSper is enhanced by the membrane hyperpolarization mediated by Slo3, a K(+) channel also present in the sperm tail. To date, no transmitter-mediated currents have been reported in sperm and no currents have been detected in the head or midpiece of mature spermatozoa. We screened a number of neurotransmitters and biomolecules to examine their ability to induce ion channel currents in the whole spermatozoa. Surprisingly, we find that none of the previously reported neurotransmitter receptors detected by antibodies alone are functional in mouse spermatozoa. Instead, we find that mouse spermatozoa have a cation-nonselective current in the midpiece of spermatozoa that is activated by external ATP, consistent with an ATP-mediated increase in intracellular Ca(2+) as previously reported. The ATP-dependent current is not detected in mice lacking the P2X2 receptor gene (P2rx2(-/-)). Furthermore, the slowly desensitizing and strongly outwardly rectifying ATP-gated current has the biophysical and pharmacological properties that mimic heterologously expressed mouse P2X2. We conclude that the ATP-induced current on mouse spermatozoa is mediated by the P2X2 purinergic receptor/channel. Despite the loss of ATP-gated current, P2rx2(-/-) spermatozoa have normal progressive motility, hyperactivated motility, and acrosome reactions. However, fertility of P2rx2(-/-) males declines with frequent mating over days, suggesting that P2X2 receptor adds a selection advantage under these conditions.
精子细胞在上升到女性生殖道的过程中获得超激活的运动能力,这使它们能够克服障碍并穿透围绕卵子的卵丘和透明带。这种增强的运动能力需要通过精子中的阳离子通道(CatSper)钙离子选择性离子通道进入 Ca(2+)。Slo3 介导的膜超极化增强了 CatSper 中的 Ca(2+)进入,Slo3 也是存在于精子尾部的一种 K(+)通道。迄今为止,尚未在精子中报道递质介导的电流,也未在成熟精子的头部或中段检测到电流。我们筛选了许多神经递质和生物分子,以检查它们诱导整个精子中离子通道电流的能力。令人惊讶的是,我们发现,通过抗体单独检测到的以前报道的神经递质受体中,没有一种在小鼠精子中具有功能。相反,我们发现,小鼠精子中段有一种阳离子非选择性电流,可被外部 ATP 激活,与先前报道的 ATP 介导的细胞内 Ca(2+)增加一致。在缺乏 P2X2 受体基因(P2rx2(-/-))的小鼠中,未检测到 ATP 依赖性电流。此外,缓慢脱敏和强烈外向整流的 ATP 门控电流具有与异源表达的小鼠 P2X2 相似的生物物理和药理学特性。我们得出结论,ATP 诱导的电流是通过 P2X2 嘌呤能受体/通道介导的。尽管丧失了 ATP 门控电流,但 P2rx2(-/-) 精子具有正常的渐进运动、超激活运动和顶体反应。然而,随着多天频繁交配,P2rx2(-/-) 雄性的生育能力下降,这表明在这些条件下,P2X2 受体增加了选择优势。