Murine SPAM1 is secreted by the estrous uterus and oviduct in a form that can bind to sperm during capacitation: acquisition enhances hyaluronic acid-binding ability and cumulus dispersal efficiency.
Sperm uptake of epididymal sperm adhesion molecule 1 (SPAM1) in vitro has recently been shown to be a marker of sperm maturation, since acquisition of this surface hyaluronidase increases cumulus dispersal efficiency. Here, we demonstrate that this glycosyl phosphatidylinositol-linked sperm antigen, previously shown to be expressed during estrous in the female reproductive tract, is secreted in the uterine and oviductal fluids (ULF and OF respectively) in a 67 kDa form, which can bind to sperm. We show that it can be acquired by caudal sperm from Spam1 null, Spam1-deficient mutant, and wild-type (WT) mice in vitro during incubation in ULF or OF at 37 degrees C, as detected by immunocytochemistry and flow cytometry. SPAM1 binding after ULF incubation was localized predominantly to the acrosome and the mid-piece of the flagella of Spam1 null sperm in a pattern identical to that of WT sperm. After ULF incubation, WT sperm demonstrated a significantly (P<0.001) enhanced hyaluronic acid-binding ability, and the involvement of SPAM1 in this activity was shown by a significant (P<0.001) decrease in binding when sperm were exposed to SPAM1 antiserum-inhibited ULF. Importantly, when Spam1 null sperm were exposed to ULF with SPAM1 accessible (in the presence of pre-immune serum) or inaccessible (in the presence of SPAM1 antiserum) for uptake, there was a significant difference in cumulus dispersal efficiency. Taken together, these results suggest that in the sperm surface remodeling that occurs prior to and during capacitation, the fertilizing competence of sperm is increased via acquisition of SPAM1, and likely other hyaluronidases, from the female tract.
Griffiths
Genevieve S
GS
Department of Biological Sciences, University of Delaware, 267 MKL, Newark, Delaware 19716, USA.
Miller
Kimberly A
KA
Galileo
Deni S
DS
Martin-DeLeon
Patricia A
PA
D000818:Animals; D015153:Blotting, Western / Q000379:methods; D015815:Cell Adhesion Molecules / Q000032:analysis / Q000378:metabolism*; D004971:Estrus / Q000502:physiology*; D005187:Fallopian Tubes / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry; D006820:Hyaluronic Acid / Q000378:metabolism; D006821:Hyaluronoglucosaminidase / Q000032:analysis / Q000378:metabolism*; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D011485:Protein Binding; D013075:Sperm Capacitation / Q000502:physiology; D013084:Sperm-Ovum Interactions / Q000502:physiology; D013094:Spermatozoa / Q000378:metabolism; D014599:Uterus / Q000378:metabolism*
D015815:Cell Adhesion Molecules; D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase; C066852:hyaluronidase PH-20