Analysis of amplification and association polymorphisms in the bovine beta-defensin 129 (BBD129) gene revealed its function in bull fertility.


Journal

Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288

Informations de publication

Date de publication:
09 11 2022
Historique:
received: 02 06 2022
accepted: 03 11 2022
entrez: 9 11 2022
pubmed: 10 11 2022
medline: 15 11 2022
Statut: epublish

Résumé

β-defensins are adsorbable on the sperm surface in the male reproductive tract (MRT) and enhance sperm functional characteristics. The beta-defensin 129 (DEFB129) antimicrobial peptide is involved in sperm maturation, motility, and fertilization. However, its role in bovine fertility has not been well investigated. This study examines the relationship between the bovine BBD129 gene and Bos indicus x Bos taurus bull fertility. The complete coding sequence of BBD129 mRNA was identified by RNA Ligase Mediated-Rapid Amplification of cDNA End (RLM-RACE) and Sanger sequencing methodologies. It consisted of 582 nucleotides (nts) including 5' untranslated region (UTR) (46nts) and 3'UTR (23nts). It conserves all beta-defensin-like features. The expression level of BBD129 was checked by RT-qPCR and maximal expression was detected in the corpus-epididymis region compared to other parts of MRT. Polymorphism in BBD129 was also confirmed by Sanger sequencing of 254 clones from 5 high fertile (HF) and 6 low fertile (LF) bulls at two positions, 169 T > G and 329A > G, which change the S57A and N110S in the protein sequence respectively. These two mutations give rise to four types of BBD129 haplotypes. The non-mutated TA-BBD129 (169 T/329A) haplotype was substantially more prevalent among high-fertile bulls (P < 0.005), while the double-site mutated GG-BBD129 (169 T > G/329A > G) haplotype was significantly more prevalent among low-fertile bulls (P < 0.005). The in silico analysis confirmed that the polymorphism in BBD129 results in changes in mRNA secondary structure, protein conformations, protein stability, extracellular-surface availability, post-translational modifications (O-glycosylation and phosphorylation), and affects antibacterial and immunomodulatory capabilities. In conclusion, the mRNA expression of BBD129 in the MRT indicates its region-specific dynamics in sperm maturation. BBD129 polymorphisms were identified as the deciding elements accountable for the changed proteins with impaired functionality, contributing to cross-bred bulls' poor fertility.

Identifiants

pubmed: 36352091
doi: 10.1038/s41598-022-23654-3
pii: 10.1038/s41598-022-23654-3
pmc: PMC9646896
doi:

Substances chimiques

beta-Defensins 0
3' Untranslated Regions 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

19042

Subventions

Organisme : Bill and Melinda Gates Foundation
ID : INV-008501-OPP1154401

Informations de copyright

© 2022. The Author(s).

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Auteurs

Subhash Solanki (S)

Animal Genomics Lab, National Dairy Research Institute, Karnal, Haryana, 132001, India.

Poonam Kashyap (P)

Animal Genomics Lab, National Dairy Research Institute, Karnal, Haryana, 132001, India.

Syed Azmal Ali (SA)

Cell Biology and Proteomics Lab, Animal Biotechnology Centre, National Dairy Research Institute, Karnal, 132001, Haryana, India.

Vijay Kumar (V)

NMR-2 Lab, National Institute of Immunology, New Delhi, 110076, India.

Ashutosh Vats (A)

Animal Genomics Lab, National Dairy Research Institute, Karnal, Haryana, 132001, India.

Martina Pukhrambam (M)

Animal Genomics Lab, National Dairy Research Institute, Karnal, Haryana, 132001, India.

Rakesh Kumar (R)

Animal Genomics Lab, National Dairy Research Institute, Karnal, Haryana, 132001, India.

Sachinandan De (S)

Animal Genomics Lab, National Dairy Research Institute, Karnal, Haryana, 132001, India.

Tirtha Kumar Datta (TK)

Animal Genomics Lab, National Dairy Research Institute, Karnal, Haryana, 132001, India. tirthadatta@gmail.com.
ICAR- Central Institute for Research on Buffaloes, Hisar, 125001, India. tirthadatta@gmail.com.

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