Expression of Toll-like receptors and avian β-defensins and their changes in response to bacterial components in chicken sperm.

The aim of the present study was to examine whether chicken sperm express mRNA of Toll-like receptors (TLR) and avian β-defensins (avBD), and whether the expression of avBD in the sperm are regulated by the interaction of TLR-2 and TLR-4 with their corresponding ligands. Total RNA was extracted from fresh sperm and mRNA expression for TLR and avBD were examined by reverse-transcription PCR. Changes in the expression of avBD-5, avBD-9, avBD-10, and avBD-12 in the sperm after incubation with lipopolysaccharide (LPS; TLR-4 ligand) or lipopeptide (Pam3CSK4; TLR-2 ligand) were examined by real-time PCR. Among 8 types of TLR, PCR products of 7 types, namely, TLR-2 to TLR-5, TLR-7, TLR-15, and TLR-21, were identified in the sperm. The sperm also expressed cluster of differentiation 14 (CD14), which is required for the binding of LPS to TLR-4. Out of 14 avBD types reported in the chicken, a total of 9 types, namely, avBD-1, avBD-3, avBD-5, and avBD-7 to avBD-12 were expressed by the sperm, whereas avBD-5, avBD-9, avBD-10, and avBD-12 showed dense signals on the gel. Expression levels of avBD-5, avBD-9, avBD-10, and avBD-12 were significantly increased in the sperm stimulated with LPS, and that of avBD-5 was also enhanced by Pam3CSK4. These results suggest that avian sperm have the ability of the innate host defense to protect themselves from infection in the male and female genital tracts.