Seahorses represent the only known group of animals with male pregnancy. Seahorses have small testis that produce a limited quantity of sperm. To date, the response of this immune-privileged organ to pathogenic infections has not been reported. β-defensin (BD) is an important innate immune defence factor against pathogens in vertebrate testis. To elucidate its immunoprotection in seahorse testis, we identified the Hippocampus erectus β-defensin (HeBD) sequence in its genome via phylogenetic tree and protein-sequence structure analysis. Gene-expression analysis showed that HeBD was highly expressed in the seahorse testis and was significantly upregulated after bacterial infection, indicating that HeBD expression was related to testicular immune responses. Furthermore, antibacterial activity testing demonstrated that the mature HeBD peptide exhibited broad-spectrum aggregation activity but only moderate antibacterial activity. We found that the mature HeBD mature significantly neutralised bacterial endotoxin activity. In conclusion, our results imply that HeBD serves an immunoprotective role in seahorse testis.
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