During mammalian fertilization, the contact between sperm and egg triggers increases in intracellular Ca² concentration ([Ca² ]_i) in sperm. Voltage-gated Ca² channels (Ca_Vs) are believed to mediate the initial phase of [Ca² ]_i increases in sperm induced by egg coat (zona pellucida [ZP]) glycoproteins, while store depletion-activated Ca² entry is thought to mediate the sustained phase. Using patch-clamp recording and Ca² imaging, we show herein that Ca_V channel currents, while found in spermatogenic cells, are not detectable in epididymal sperm and are not essential for the ZP-induced [Ca² ]_i changes. Instead, CATSPER channels localized in the distal portion of sperm (the principal piece) are required for the ZP-induced [Ca² ]_i increases. Furthermore, the ZP-induced [Ca² ]_i increase starts from the sperm tail and propagates toward the head.