This Article Accepted Manuscript (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Google Scholar Articles by Yudin, A. Articles by Bevins, C. PubMed PubMed Citation Articles by Yudin, A. Articles by Bevins, C.

β-defensin 22 is a major component of the mouse sperm glyocalyx

A Yudin, Center for Health and the Environment, University of California, Davis, Davis, United States
T Tollner, Center for Health and the Environment, University of California, Davis, Davis, 95616, United States
C Treece, Center for Health and the Environment, University of California, Davis, Davis, United States
R Kays, Department of Microbiology and Immunology, School of Medicine, University of California, Davis, United States
G Cherr, Bodega Marine Lab, UC Davis, Bodega Bay, United States
J Overstreet, Center for Health and the Environment, University of California, Davis, Davis, United States
C Bevins, Department of Microbiology and Immunology, School of Medicine, University of California, Davis, United States

Correspondence: Ted Tollner, Email: tltollner{at}ucdavis.edu

Abstract

Surface components of sperm isolated from the cauda epididymidis were stabilized by whole sperm fixation for immunization of rabbits. The resulting immunoglobulins (Igs) recognized a single protein of 130kDa (non-reduced) or 54-57kDa (reduced) on Western blots of cauda sperm. Igs recognized the same 54-57 kDa protein band on whole tissue blots of the corpus and cauda epididymides and vas deferens. No immunoreactive bands were detected on blots of the prostate, seminal vesicles, testes, caput epididymis, or any of various non-reproductive tissues. Removal of sperm from the vas deferens prior to blotting eliminated the detection of the sperm antigen. Antibodies raised to synthetic peptides, identical in amino acid sequence to two unique spans of beta-defensin 22, recognized the same 130/54-57kDa antigen on Western blots of both caudal sperm and the purified antigen isolated with the anti-sperm Ig. From indirect immunoflourescence, both the anti-sperm and anti-peptide Igs appeared to localize to the entire sperm surface, a pattern confirmed at the ultrastructural level. Real-time PCR identified the corpus epididymis as the major site of expression of beta-defensin 22, with negligible expression in the testes, caput epididymis and vas deferens. Immuno-staining of epididymal sections showed beta-defensin 22 being released into the lumen at the distal caput / proximal corpus, with sperm becoming intensely coated with beta-defensin 22 as they reached the distal corpus. Most uterine sperm recovered from mice 4hrs following copulation exhibited beta-defensin 22 coating the entire sperm surface. By contrast, some sperm recovered from the oviduct and cumulus extracellular matrix showed loss of beta-defensin 22 from the sperm head.