Genetic variation in the BLM gene and its expression in the ovaries is closely related to kidding number in goats.

Bloom (BLM) helicase plays an important role in DNA replication and the maintenance of genome integrity. BLM protein deficiency, which plays a vital role in the sperm-egg union and germ-cell development during reproduction, can lead to severe DNA damage in goats. However, the effect of BLM protein deficiency on goat litter size has not been reported. Herein, we studied the association between the genetic variation in the BLM gene and the number of kids per litter in Guizhou white goats. We explored differences in the expression of the BLM protein in the follicles of single and multi-kid nanny goats. We also analyzed the effects of dysregulated BLM gene expression on the proliferation and apoptosis of ovarian granulosa cells and the expression of genes related to follicle development in goats. Five single nucleotide polymorphism (SNP) loci, including the non-synonymous mutations g.38179 A > G, g.40626 G > C and g.89621 T > G; the intron synonymous mutation g.56961 G > A and the exon synonymous mutation g.65796 C > T were found in the BLM gene. All SNPs loci were in Hardy-Weinberg equilibrium, and correlation analysis showed that the g.65796 C > T and g.89621 T > G loci polymorphism was strongly associated with litter size in the first three litters (P < 0.05). The diplogenotype Hap 2/2 (AAGGAACCTT) showed no significant difference in litter size between different births, indicating that the diploid genotype is stable in different litter sizes. Bioinformatics analysis showed that three non-synonymous mutation loci (p.T488A, p.A662S, and p.S1373A) could affect BLM protein stability, and mutations in p.T488A and p.S1373A led to changes in amino acid polarity and associated interactions. qPCR results showed that the expression level of the BLM gene in the uterus and ovaries of TT genotype nanny goats was significantly higher than that of GG genotype nanny goats. Indirect immunofluorescence assay (IF) showed that the BLM protein was significantly overexpressed in both the primordial and growing follicles of nanny goats with multiple kids (P < 0.01). Disrupting BLM gene expression in the ovarian granulosa cells down-regulated the expression of the Cyp19A1 gene. It also significantly inhibited the proliferation of follicles and induces early apoptosis of the granulosa cells. These findings confirm that polymorphism in the BLM gene is closely related to the littering traits of Guizhou white goats, and it affects the reproductive performance of nanny goats by regulating the development of the oocytes and granulosa cells. This work provides new evidence on the regulatory effect of the BLM gene on the litter size of nanny goats.

A novel nonsynonymous SNP in the OLR1 gene associated with litter size in Guizhou white goats.

Oxidized low-density lipoprotein receptor-1 (OLR1) encodes a low-density lipoprotein receptor belonging to the C-type lectin superfamily, which is closely related to reproduction. OLR1 is associated with fecundity in Awassi sheep. However, its effect on litter size has not been investigated in goats. In this study, OLR1 sequences and their mRNA expression levels in the gonadal axis of Guizhou white goats were evaluated to investigate the relationship between gene polymorphisms and litter size. In addition, the potential effects of a nonsynonymous substitution were evaluated using a bioinformatics approach. The expression levels of OLR1 were highest in the uterus of mothers with multiple kids and highest in the ovaries of mothers with single kids. OLR1 mRNA expression levels in the ovaries of mothers with single kids were two times higher than in the ovaries of mothers with multiple kids. The sequencing results revealed five SNPs in OLR1; however, only g.294 T > A, g.2260 T > C, and g.2268 C > T were significantly associated with litter size (P < 0.05). Linkage disequilibrium was detected between g.2260 T > C and g.2268 C > T (r2 = 0.322, D' = 0.6). Additionally, goats with the Hap 1/1 diplotype had a greater litter size than others (P < 0.05). g.2260 T > C was a nonsynonymous mutation that resulted in the replacement of valine with alanine at the amino acid residue 54 of the OLR1 protein. Bioinformatic analyses revealed that the p.V54A locus was relatively conserved in cloven-hoofed species. Mutations at this locus could change the local conformation and reduce the stability of OLR1, affecting its half-life and the litter size of the nanny goat. These findings confirm that OLR1 affects goat kidding traits and provide a novel insight into the regulatory mechanism underlying the effect of OLR1 on litter size.