title: Mice lacking two sperm serine proteases, ACR and PRSS21, are subfertile, but the mutant sperm are infertile in vitro.
abstract: Although sperm serine protease and proteasome have long been believed to play an important role in the fertilization process, the molecular mechanism is still controversial. In this study, we have produced double-knockout mice lacking two sperm serine proteases, ACR and PRSS21, to uncover the functional role of the trypsinlike activity in fertilization. The double-knockout male mice were subfertile, likely owing to the incompleteness of fertilization in the oviductal ampulla. Despite male subfertility, the mutant epididymal sperm exhibited the inability to undergo acrosomal exocytosis on the zona pellucida (ZP) surface and to traverse the ZP, thus resulting in the failure of fertilization in vitro. The double-knockout epididymal sperm were also defective in penetration through the cumulus matrix to reach the ZP. When epididymal sperm were artificially injected into the uterus of wild-type mice, the 2-cell embryos, which had previously been fertilized by double-knockout sperm, were recovered at a low but significant level. The mutant epididymal sperm were also capable of fertilizing the oocytes in the presence of uterine fluids in vitro. These data demonstrate that the trypsinlike protease activity of ACR and PRSS21 is essential for the process of sperm penetration through the cumulus matrix and ZP in vitro, and suggest that the female reproductive tract partially compensates for the loss of the sperm function. We therefore conclude that the sperm trypsinlike activity is still important but not essential for fertilization in vivo in the mouse.
authors.affiliation: Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba Science City, Ibaraki, Japan.
authors.forename: Natsuko
authors.initials: N
authors.lastname: Kawano
authors.forename: Woojin
authors.initials: W
authors.lastname: Kang
authors.forename: Misuzu
authors.initials: M
authors.lastname: Yamashita
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Koga
authors.forename: Taiga
authors.initials: T
authors.lastname: Yamazaki
authors.forename: Tamako
authors.initials: T
authors.lastname: Hata
authors.forename: Kenji
authors.initials: K
authors.lastname: Miyado
authors.forename: Tadashi
authors.initials: T
authors.lastname: Baba
mesh_terms: D000176:Acrosin / Q000235:genetics / Q000378:metabolism*; D020101:Acrosome Reaction / Q000502:physiology; D000818:Animals; D015153:Blotting, Western; D054885:Cumulus Cells / Q000502:physiology; D005260:Female; D005298:Fertility / Q000502:physiology*; D005306:Fertilization / Q000502:physiology*; D058851:GPI-Linked Proteins / Q000235:genetics / Q000378:metabolism; D007248:Infertility, Male / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D018345:Mice, Knockout; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D013084:Sperm-Ovum Interactions / Q000502:physiology; D013094:Spermatozoa / Q000502:physiology*; D015044:Zona Pellucida / Q000502:physiology
chemical_list: D058851:GPI-Linked Proteins; C456340:Prss21 protein, mouse; D012697:Serine Endopeptidases; D000176:Acrosin