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Outer Dense Fiber Proteins Are Dominant Postobstruction Autoantigens in Adult Lewis Rats¹

^a Department of Cell Biology and the Center for Recombinant Gamete Contraceptive Vaccinogens, and ^b the W.M. Keck Biomedical Mass Spectrometry Laboratory, University of Virginia, Charlottesville, Virginia 22908 ^c Department of Anatomy & Cell Biology, Queens University, Kingston, Ontario, Canada K7L 3N6

ABSTRACT

Obstruction of the male reproductive tract commonly results in generation of antisperm autoantibodies. However, only a few of the sperm autoantigens recognized by these antibodies have been characterized. To identify postobstruction rat sperm autoantigens, sperm proteins were separated by two-dimensional(2-D) gel electrophoresis. Spots corresponding to proteins that were stained by at least 50% of postvasectomy rat sera on 2-D Western blots were removed from polyacrylamide gels and microsequenced by tandem mass spectrometry. From a total of 21 spots, 12 contained peptides that matched solely to either of two outer dense fiber proteins, odf1 or odf2. Six additional spots contained peptides comprising odf1 or odf2 and were accompanied by peptides representing other proteins. Only three spots lacked outer dense fiber peptides but did contain sequences of other known proteins. The results indicate that the outer dense fiber proteins odf1 and odf2 are dominant postobstruction autoantigens because they were detected in the majority of the immunoreactive protein spots examined. Possible explanations for this observation include the abundance of outer dense fiber proteins in spermatozoa, slow solubility, which may provide a sustained supply of antigen, and testis-specific expression during spermiogenesis.

FOOTNOTES

First decision: 12 September 2000.

¹ Supported by the National Institutes of Health (NIDDK P50 DK45179, NICHD HD U54-29099), the Fogarty International Center (D43 TW/HD 00654), and the NICHD/NIH through cooperative agreement U54 HD28934 as part of the Specialized Cooperative Centers Program in Reproduction Research; the Andrew W. Mellon Foundation; and Schering AG.

² Correspondence: Charles J. Flickinger, Department of Cell Biology, School of Medicine, University of Virginia Health System, P.O. Box 800732, Charlottesville, VA 22908-0732. FAX: 804 982 3912;cjf{at}virginia.edu

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