Dual Sensing of Physiologic pH and Calcium by EFCAB9 Regulates Sperm Motility

Varying pH of luminal fluid along the female reproductive tract is a physiological cue that modulates sperm motility. CatSper is a sperm-specific, pH-sensitive calcium channel essential for hyperactivated motility and male fertility. Multi-subunit CatSper channel complexes organize linear Ca²⁺ signaling nanodomains along the sperm tail. Here, we identify EF-hand calcium-binding domain-containing protein 9 (EFCAB9) as a bifunctional, cytoplasmic machine modulating the channel activity and the domain organization of CatSper. Knockout mice studies demonstrate that EFCAB9, in complex with the CatSper subunit, CATSPERζ, is essential for pH-dependent and Ca²⁺-sensitive activation of the CatSper channel. In the absence of EFCAB9, sperm motility and fertility is compromised, and the linear arrangement of the Ca²⁺ signaling domains is disrupted. EFCAB9 interacts directly with CATSPERζ in a Ca²⁺-dependent manner and dissociates at elevated pH. These observations suggest that EFCAB9 is a long-sought, intracellular, pH-dependent Ca²⁺ sensor that triggers changes in sperm motility. A pH-dependent calcium sensor enables modulation of sperm motility in response to changing conditions along the female reproductive tract.