Detection of Bovine TMEM95 p.Cys161X Mutation in 13 Chinese Indigenous Cattle Breeds

Abstract

Chinese indigenous cattle breeds have abundant genetic resources, which are valuable for the molecular breeding of cattle around the world. Thus, identifying important candidate genes and their genetic markers is very important for cattle molecular breeding. A previous study found that a nonsense mutation (rs378652941, c.483C > A, p.Cys161X) in the bovine transmembrane protein 95 gene (TMEM95) seriously reduced the reproductive performance in bulls, but few studies have detected this mutation in Chinese indigenous cattle breeds. Since the mutation c.483C > A may serve as a potential genetic marker for selecting higher fertility bulls, in the present study, using tetra-primer amplification refractory mutation system PCR (T-ARMS-PCR), forced PCR restriction fragment length polymorphism (forced PCR-RFLP), and DNA sequencing methods, the mutation c.483C > A was detected in 765 individuals from 13 Chinese cattle breeds. However, the results showed that this mutation did not exist at this locus in our analyzed breeds. Interestingly, we identified a newly frameshift insertion/deletion (indel) mutation (NC_037346.1: g.27056998_27057000delCT) in the bovine TMEM95 gene in 11 cattle breeds, which changed the location of the termination codon and changed the 16 amino acids in the C-terminal to 21 amino acids. Combined with previous studies, our study provides evidence that in Chinese cattle breeds the mutation c.483C > A cannot be used as a genetic marker in molecular breeding.

The Role of Sperm Proteins IZUMO1 and TMEM95 in Mammalian Fertilization: A Systematic Review

“…The results showed that eight articles studied TMEM95 ( Table 1 ). The main objectives of these articles were to molecularly characterize TMEM95 [ 20 , 22 , 31 , 35 ] or to determine the cause or effect of a certain mutation, including nonsense mutations [ 23 , 24 ], synonymous mutations [ 32 ] and insertion/deletion mutations [ 36 ].…”

Development of alternative diagnosis of HH1, HH3, HH5 and HCD fertility haplotypes and subfertility syndrome in cattle

“…The presence of a nonsense mutation (rs378652941, c.483C>A, p.Cys161X) in the bovine transmembrane protein in the VI exon part of the TMEM95 gene seriously reduced the reproductive capacity of bulls. Since the c.483C>A mutation can serve as a potential genetic marker for the selection of bulls with higher fertility, in the present study, T‐ARMS‐PCRs and PCR‐RFLP analysis methods were used to identify genetic variants at the TMEM95 gene locus, DNA sequencing method; however, the authors did not reveal mutations c.483C>A in the studied animals carriers (Zhang et al., 2019).…”

Human sperm TMEM95 binds eggs and facilitates membrane fusion

“…Sperm IZUMO1 (1) and egg JUNO (2) mediate the only known cell surface interaction between mammalian gametes. Recent reports suggested that Tmem95 (encoding transmembrane protein 95) mutant cattle exhibit impaired male fertility (3, 4); Tmem95 knockout mice show male-specific sterility, producing sperm that can bind to, but do not fuse with eggs (5, 6). Tmem95 encodes a sperm acrosomal membrane protein, which re-localizes to the equatorial segment of the sperm head (3, 6) where membrane fusion with the egg takes place (7, 8).…”