Frequency of an X-Linked Maternal Variant of the Bovine FOXP3 Gene Associated with Infertility in Different Cattle Breeds: A Pilot Study

Abstract

Immune adaptation plays an essential role in determining pregnancy, which has been shown to be dependent on sufficient immunological tolerance mediated by FOXP3+ regulatory T cells. Recently, an X-linked maternal single-nucleotide polymorphism (SNP), located 2175 base pairs upstream of the start codon in the bovine FOXP3 gene (NC_037357.1: g.87298881A>G, rs135720414), was identified in Japanese Black (JB: Bos taurus) cows in association with recurrent infertility. However, with the exception of JB cows, the frequency of this SNP has yet to be studied in other cow populations. In this study, we thus aimed to evaluate the frequency of this SNP in different cow breeds. Between 2018 and 2021, a total of 809 DNA samples were obtained from 581 JB, 73 Holstein Friesian (HF: B. taurus), 125 Korean Hanwoo (KH: B. taurus coreanae), and 30 Indonesian Madura (IM: a crossbreed between B. indicus and B. javanicus) cows, which were genotyped using a TaqMan probe-based real-time polymerase chain reaction assay designed in this study. The frequency of the G allele was found to be relatively high in local IM (0.700), moderate in dairy HF (0.466), and low in beef JB (0.250) and KH (0.112) cows, with differences in the frequencies between each group being shown to be statistically significant (p < 0.005) using Fisher’s exact test. The results obtained in this study indicate that the G allele frequencies of the identified the SNP differ markedly in different breeds of taurine and indicine cattle. Given these findings, it would thus be important to evaluate the relationships between high frequencies of the G allele and infertility in different breeds.

Novel Approach for Evaluating Pregnancy-Associated Glycoprotein and Inflammation Markers during the Postpartum Period in Holstein Friesian Cows

“…In recent years, uncovering the role of maternal immune regulatory cells and their regulatory genes that may also contribute to infertility has gained increasing attention in veterinary research. In particular, the development, differentiation, and immunosuppressive functions of T-regs have been the focus of numerous studies since the identification of FOXP3 [41,42]. We detected cows with FOXP3 variants G/G (n = 4) and A/G (n = 13), but not A/A (n = 2; Table 3).…”

“…We detected cows with FOXP3 variants G/G (n = 4) and A/G (n = 13), but not A/A (n = 2; Table 3). Recently, we reported the frequency of the G allele of the FOXP3 variant in four cattle breeds (Japanese Black, Holstein, Korean Hanwoo, and Indonesian Madura) and found that the frequency in Holstein cattle was 0.466 (n = 73) [42]. Interestingly, significant differences (p = 0.017) in follicular cysts (A/A: 0% and G/G: 100%) were observed among the three groups; however, the limited number of cows in the present study prevents this result from being fully generalizable.…”

“…Deoxyribonucleic acid (DNA) was extracted from the disks punched out from bloodimpregnated FTA cards following appropriate treatment, as described previously [46]. Genotyping was performed as described previously [42]. Briefly, the primers and TaqMan minor groove binder (MGB) probes used for real-time polymerase chain reaction (RT-PCR) assays were designed based on the sequence of bovine FOXP3 (NCBI Reference Sequence NC_037357.1).…”

“…These mutations may have a detrimental influence on the development of maternal T-reg cells, which may encourage the activation of effector T cells specific to the fetus, leading to infertility [41]. FOXP3 variants (NC_037357.1: g.87298881A>G, rs135720414) may affect T-reg cell function and are related to negative reproductive performance, such as that in repeat breeders of Japanese Black cattle [41], with a higher G allele frequency of 0.466 in Holstein Friesian dairy herds [42]. No studies have reported a relationship between the postpartum clinical parameters affecting reproductive efficacy and FOXP3 variants.…”