Partial Deletion of Chromosome 8 β-defensin Cluster Confers Sperm Dysfunction and Infertility in Male Mice

Abstract

β-defensin peptides are a family of antimicrobial peptides present at mucosal surfaces, with the main site of expression under normal conditions in the male reproductive tract. Although they kill microbes in vitro and interact with immune cells, the precise role of these genes in vivo remains uncertain. We show here that homozygous deletion of a cluster of nine β-defensin genes ( DefbΔ9) in the mouse results in male sterility. The sperm derived from the mutants have reduced motility and increased fragility. Epididymal sperm isolated from the cauda should require capacitation to induce the acrosome reaction but sperm from the mutants demonstrate precocious capacitation and increased spontaneous acrosome reaction compared to wild-types but have reduced ability to bind the zona pellucida of oocytes. Ultrastructural examination reveals a defect in microtubule structure of the axoneme with increased disintegration in mutant derived sperm present in the epididymis cauda region, but not in caput region or testes. Consistent with premature acrosome reaction, sperm from mutant animals have significantly increased intracellular calcium content. Thus we demonstrate in vivo that β-defensins are essential for successful sperm maturation, and their disruption leads to alteration in intracellular calcium, inappropriate spontaneous acrosome reaction and profound male infertility.

Author Summary

β-defensins are small molecules, considered primarily to be antimicrobials and important in the first defence response to invading organisms. They are predominantly produced at surfaces in contact with the outside environment and these include skin, airway and reproductive tract. We show here that when we delete from the mouse a subset of nine β-defensin genes, surprisingly the main consequence is that the male mice are completely infertile. When normal sperm leave the male and enter the female reproductive tract they are triggered to undergo a reaction that alters the membrane properties of the sperm and allows fertilisation. We show here that sperm isolated from the male mice, that no longer make these β-defensins, are prematurely ready to fertilise an egg. It is far too early for this to happen and as a consequence the sperm are severely reduced in their ability to move and have a major defect in the structure of their tail. We provide evidence that the reason this has happened is due to a dysregulation of calcium transport. This work is important for understanding defensin gene function in a living organism and may enable the design of novel contraceptives with additional antibiotic ability.