The relationship of polymorphism of the FSHR and INHBA genes with the quality of ejaculate in dogs of various breeds

Sperm quality plays a crucial role in the successful implementation of breeding programs, especially in cases where artificial insemination is practiced. Males with excellent sperm quality have good fertility and can produce larger litters, and their ejaculate is more resistant to preservation. The aim of this study is to experimentally and analytically confirm the hypothesis of the association of SNP exons of candidate genes FSHR and INHBA with the sperm quality of male dogs. Through mathematical analysis of the genomic association data (GWAS) we obtained, it was possible to identify genomic regions and individual variations associated with production traits in dogs. When selecting primers for the amplification of the 2 exons of the FSHR gene and the 9 and 10 exons of the INHBA gene, the reference sequence of the domestic dog genome (Canis familiaris) was used from the NCBI international genetic database. The primer selection was carried out using the Primer-BLAST application integrated into NCBI (https://www.ncbi.nlm.nih.gov/). The primers were designed by Sintol LLC upon individual request. Amplification was performed on a C -1000 thermocycler (BioRad). Sequencing of amplified fragments was performed using nanopore sequencing technology on a MinION instrument. A Rapid Sequencing Kit was used to prepare DNA libraries. Variants were determined using the longshot program (v. 1.0.0). The results of the analysis demonstrate a statistically significant effect (p<0.01) of the INHBA2 and FSHR genes, which contain SNP mutations in the studied loci, on the quality of ejaculate in dogs. These findings contribute to the little-studied field of companion animal genetics. The data obtained opens up new opportunities for diagnosing the causes of male infertility. The identified patterns set a clear direction for further work: validation on an expanded sample and the search for new significant genetic markers.

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