Role of sperm surface arylsulfatase A in mouse sperm-zona pellucida binding.
We have previously described the zonae pellucidae (ZP) binding ability of a pig sperm surface protein, P68. Our recent results on peptide sequencing of 3 P68 tryptic peptides and molecular cloning of pig testis arylsulfatase A (AS-A) revealed the identity of P68 as AS-A. In this report, we demonstrate the presence of AS-A on the mouse sperm surface and its role in ZP binding. Using anti-AS-A antibody, we have shown by immunoblotting that AS-A was present in a Triton X-100 extract of mouse sperm. The presence of AS-A on the sperm plasma membrane was conclusively demonstrated by indirect immunofluorescence, immunogold electron microscopy, and AS-A's desulfation activity on live mouse sperm. The AS-A remained on the head surface of in vivo capacitated sperm, as revealed by positive immunofluorescent staining of oviductal/uterine sperm. Significantly, the role of mouse sperm surface AS-A on ZP binding was demonstrated by dose-dependent decreases of sperm-ZP binding on sperm pretreatment with anti-AS-A IgG/Fab. Furthermore, Alexa-430 conjugated AS-A bound to mouse ZP of unfertilized eggs but not to fertilized ones, and this level of binding increased and approached saturation with increasing Alexa-430 AS-A concentrations. Moreover, in vivo fertilization was markedly decreased when mouse sperm pretreated with anti-AS-A IgG were artificially inseminated into females. All of these results designated a new function for AS-A in mouse gamete interaction.
Tantibhedhyangkul
Julierut
J
Hormones/Growth/Development Group, Ottawa Health Research Institute, 725 Parkdale Avenue, Ottawa, Ontario, Canada K1Y 4E9.
Weerachatyanukul
Wattana
W
Carmona
Euridice
E
Xu
Hongbin
H
Anupriwan
Araya
A
Michaud
Dominick
D
Tanphaichitr
Nongnuj
N
D000176:Acrosin / Q000037:antagonists & inhibitors / Q000276:immunology; D020101:Acrosome Reaction / Q000502:physiology; D000818:Animals; D015153:Blotting, Western; D002553:Cerebroside-Sulfatase / Q000502:physiology*; D004591:Electrophoresis, Polyacrylamide Gel; D004822:Epididymis / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D005260:Female; D005307:Fertilization in Vitro; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008854:Microscopy, Electron; D011506:Proteins / Q000378:metabolism; D013075:Sperm Capacitation / Q000502:physiology; D013084:Sperm-Ovum Interactions / Q000187:drug effects; D013094:Spermatozoa / Q000201:enzymology* / Q000502:physiology*; D014465:Ultrasonics; D014649:Vas Deferens / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D015044:Zona Pellucida / Q000502:physiology*
D011506:Proteins; D002553:Cerebroside-Sulfatase; D000176:Acrosin