Abstract
Bone morphogenetic protein 15 (BMP15) is secreted by the mammalian oocytes and is indispensable for ovarian follicular development, ovulation, and fertility. To determine the regulation mechanism of BMP15 gene, the regulatory sequence of porcine BMP15 was investigated in this study. The cloned BMP15 promoter retains the cell-type specificity, and is activated in cells derived from ovarian tissue. The luciferase assays in combination with a series of deletion of BMP15 promoter sequence show that the −427 to −376 bp region of BMP15 promoter is the primary regulatory element, in which there are a number of transcription factor binding sites, including LIM homeobox 8 (LHX8), newborn ovary homeobox gene (NOBOX), and paired-like homeodomain transcription factor 1 (PITX1). Determination of tissue-specific expression reveals that LHX8, but not PITX1 and NOBOX, is exclusively expressed in pig ovary tissue and is translocated into the cell nuclei. Overexpression of LHX8 in Chinese hamster ovary (CHO) cells could significantly promote BMP15 promoter activation. This study confirms a key regulatory element that is located in the proximal region of BMP15 promoter and is regulated by the LHX8 factor.
Highlights
Reproductive capability is a vital economic factor for sows, for instance the Chinese Taihu breed farrows at least five more piglets per litter than the Large White breed [1]
Bioinformatics analysis showed that several potential binding sites for the reproduction related factors, including germ cell nuclear factor (GCNF), LIM homeobox 8 (LHX8), newborn ovary homeobox gene (NOBOX), and paired-like homeodomain transcription factor 1 (PITX1), were found in the promoter sequence of bone morphogenetic protein 15 (BMP15) gene, indicating that BMP15 may be a downstream target of those transcription factors (Figure 1)
We found that there was no typical TATA box element in the proximal region of porcine BMP15 promoter
Summary
Reproductive capability is a vital economic factor for sows, for instance the Chinese Taihu breed farrows at least five more piglets per litter than the Large White breed [1]. Many studies have revealed that oocytes can produce and secrete growth factors that facilitate follicular development [2,3]. Bone morphogenetic protein 15 (BMP15) and growth and differentiation factor 9. (GDF9), belonging to the transforming growth factor β (TGF-β) superfamily, are produced by oocytes [4,5]. These two factors participate in the different stages of follicular development and influence the final events of embryo maturation and ovulation [6,7]. Bmp homozygous mutant causes subfertile, showing the ovulation defects and reduction of oocytes [8].
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