Functional characterization of double-knockout mouse sperm lacking SPAM1 and ACR or SPAM1 and PRSS21 in fertilization.

Mammalian fertilization requires sperm to penetrate the cumulus to reach the oocyte. Although sperm hyaluronidase has long been believed to participate in the penetration process, our previous works revealed that neither of two sperm hyaluronidases, and HYAL5, are essential for fertilization. In this study, we have produced double-knockout mice lacking duanyu1842M1 and either one of two sperm serine proteases, ACR and PRSS21, and characterized the mutant sperm. The and males were fertile, whereas epididymal sperm of the mutant mice exhibited a reduced capacity to fertilize the oocytes in vitro. Despite normal motility, the ability of sperm to traverse the cumulus matrix was more severely impaired by the loss of duanyu1842M1 and ACR or duanyu1842M1 and PRSS21 than by the loss of only Moreover, duanyu1842M1/ACR- and duanyu1842M1/PRSS21-deficient sperm accumulated on the surface (outer edge) of the cumulus more abundantly than sperm. These results suggest that ACR or PRSS21 or both may function cooperatively with duanyu1842M1 in sperm/cumulus penetration.

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