Buck (Capra hircus) genes encode new members of the spermadhesin family

Abstract

Spermadhesins are the major proteins of the boar seminal plasma and form a group of polypeptides probably involved in different steps of reproduction. In previous work, a member of the spermadhesin family from buck (Capra hircus ) seminal plasma, named BSFP, was isolated and characterized by mass spectrometry and the N-terminal was sequenced. This work aimed to found and characterize the BSFP gene and detect its expression along the male genital tract using real-time PCR and electrophoresis. The cDNAs of seminal vesicle, testis, epididymis, bulbourethral gland and ductus deferens were prepared from a mature buck. Following 3´ and 5´-end amplifications using seminal vesicle cDNA, we cloned and sequenced 4 highly similar (97 to 98%) nucleotide sequences encoding spermadhesins, named Bodhesin-1, -2, -3 and -4. All deduced amino acid sequences presented the CUB domain signature and were 49 to 52% similar to boar AWN. The Bodhesin-2 is identical to the BSFP N-terminal. After real-time PCR we noticed specific amplifications for all Bodhesins in seminal vesicle, testis, epididymis and bulbourethral gland, with the exception of Bodhesin-2 in epididymis. The amplicons presented melting temperature and size of approximately 78 ° C and 130 bp, respectively. Bodhesin expression is significantly higher in seminal vesicle when compared to the other tissues. The present work confirm that goat is the fourth mammalian species, together with pig, cattle and horse, in which spermadhesin molecules have been found. To our knowledge, this is the first report of buck spermadhesins genes using molecular cloning and expression profile.