title: Protease activation and the signal transduction pathway regulating motility in sperm from the water strider Aquarius remigis.
abstract: Many motile processes are regulated such that movement occurs only upon activation of a signaling cascade. Sperm from a variety of species are initially quiescent and must be activated prior to beating. The signaling events leading to the activation and regulation of sperm motility are not well characterized. Mature seminal vesicle sperm from the water strider Aquarius remigis are immotile in vitro, but vigorous motility is activated by trypsin. Trypsin-activated motility was blocked by pretreatment of the sperm with BAPTA-AM to chelate intracellular Ca(2+) and was partially rescued by subsequent addition of A23187 and Ca(2+). Thapsigargin stimulated motility in the absence of trypsin, suggesting that intracellular Ca(2+) stores are available. In addition, motility could be fully activated by the phosphatase inhibitor calyculin A, suggesting that the immotile state is maintained by an endogenous phosphatase and that kinase activity is required for motility. The MEK1/2 inhibitor U0126 significantly reduced trypsin activated motility, and MPM-2, an antibody which recognizes proline-directed phosphorylation by kinases such as MAPK, recognized components of the water strider sperm flagellum. Antibodies specific for the mouse protease activated receptor PAR2 recognized an antigen on the sperm flagellum. These results suggest that trypsin stimulates a Ca(2+) and MAPK mediated signaling pathway and potentially implicate a PAR2-like protein in regulating motility.
authors.affiliation: Department of Biology, University of California, Riverside, California, USA. hmiya003@biken.osaka-u.ac.jp
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Miyata
authors.forename: Catherine D
authors.initials: CD
authors.lastname: Thaler
authors.forename: Leah T
authors.initials: LT
authors.lastname: Haimo
authors.forename: Richard A
authors.initials: RA
authors.lastname: Cardullo
mesh_terms: D000818:Animals; D002118:Calcium / Q000502:physiology; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D020062:Heteroptera / Q000502:physiology*; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008297:Male; D008387:Marine Toxins; D010080:Oxazoles / Q000494:pharmacology; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011494:Protein Kinases / Q000378:metabolism; D044464:Receptor, PAR-2 / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D013081:Sperm Motility / Q000187:drug effects / Q000502:physiology*; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D019311:Staurosporine / Q000494:pharmacology; D019284:Thapsigargin / Q000494:pharmacology; D014357:Trypsin / Q000378:metabolism
chemical_list: D008387:Marine Toxins; D010080:Oxazoles; D047428:Protein Kinase Inhibitors; D044464:Receptor, PAR-2; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D019284:Thapsigargin; C059041:calyculin A; D011494:Protein Kinases; D014357:Trypsin; D019311:Staurosporine; D002118:Calcium