BMP4 Gene Expression in Small Tail Han Sheep (Ovis aries) with Different Litter Size and Association Analysis Between Its Missense Mutation and Litter Size
WEN Yu-Liang1, WANG Xiang-Yu2, GUO Xiao-Fei2, HE Xiao-Yun2, ZHAO Sheng-Guo1, *, CHU Ming-Xing2, *
1 College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China; 2 Key Laboratory of Animal Genetics, Breeding and Reproduction of Ministry of Agriculture/Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Abstract Bone morphogenetic protein (BMP)/Smad family member (SMAD) signaling pathway plays important role in the regulation of mammalian cell proliferation and differentiation. In order to study the effects of BMP4 and BMP/Smad signaling pathway on litter size of Small Tail Han (STH) sheep (Ovis aries). The present study used qRT-PCR to detect the expression of BMP4 gene and its downstream gene SMAD4 in the BMP/Smad signaling pathway in the 14 tissues (heart, liver, spleen, lung, kidney, brain, cerebellum, hypothalamus, ovary, pituitary, uterus, oviduct, thyroid, and adrenal gland). These tissues were collected from STH sheep (3 polytocous sheeps and 3 monotocous sheeps) in the follicular phase. In previous work, a missense mutation g.63454744T>G in BMP4 coding region was found by sheep genome resequencing. Then the genotypes of g.63454744T>G were detected by Sequenom MassARRAY® SNP technique in 380 STH sheep and the association between the mutation and litter size were analyzed. The results of qRT-PCR showed that these two genes expressed in all 14 tissues in follicular phase. The BMP4 gene highly expressed in spleen, liver, heart, and uterus. SMAD4 gene were highly expressed in cerebellum, hypothalamus, pituitary, and ovary. The expression of BMP4 gene in uterus (P<0.001), hypothalamus (P=0.002), pituitary (P=0.001), and ovary (P<0.001) of polytocous ewes were extremely significant higher than that of monotocous ewes. The SMAD4 gene expression in hypothalamus (P<0.001), cerebellum (P=0.001), ovary (P<0.001), pituitary (P=0.001), brain (P=0.001), uterus (P=0.007) and oviduct (P=0.005) of polytocous ewes were extremely significant higher than that of monotocous ewes. The SNP g.63454744T>G in BMP4 gene had a low polymorphism (polymorphism information content<0.25) in STH sheep. The χ2 test indicated that the locus of g.63454744T>G of BMP4 gene was under Hardy-Weinberg equilibrium (P>0.05) in this STH sheep population. The polymorphisms of g.63454744T>G in BMP4 was significantly associated with each parity litter size in STH sheep (P<0.05), and the litter size of mutant homozygotes was higher than that of wild types and heterozygotes. Therefore, it could be concluded that there was a positive correlation between BMP4 and SMAD4 gene expression and litter size in STH sheep during follicular phase, and the SNP locus of g.63454744T>G in BMP4 might have significant association with the litter size in each parity of STH sheep. The present study provides a reference basis for lambing traits selection of STH sheep.
WEN Yu-Liang,WANG Xiang-Yu,GUO Xiao-Fei, et al. BMP4 Gene Expression in Small Tail Han Sheep (Ovis aries) with Different Litter Size and Association Analysis Between Its Missense Mutation and Litter Size[J]. 农业生物技术学报, 2019, 27(1): 80-88.
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