Abstract
BMP1/TLD-related metalloproteinases play a key role in morphogenesis via the proteolytic maturation of a number of extracellular matrix proteins and the activation of a subset of growth factors of the transforming growth factor beta superfamily. Recent data indicated that BMP1 is expressed in sheep ovarian follicles and showed a protease activity. The aim of the present study was to characterize the function of the buffalo BMP1 gene in folliculogenesis. A 3195-bp buffalo BMP1 mRNA fragment was firstly cloned and sequenced, which contained a whole 2967-bp codon sequence. The multialigned results suggested that BMP1 is highly conserved among different species both at the nucleic acid and the amino acid level. BMP1 is located in the oogonium of the fetal buffalo ovary and in the granulosa cells (GCs) and the oocytes of adult ovary from the primordial to the large antral follicles. Further study showed that BMP1 promoted cell cycle and proliferation and inhibited apoptosis in IVC GCs. Adding BMP1 recombinant protein to the culture medium of the GCs increased the expression of the key cell cycle regulators such as cyclin D1 and cyclin D2 and downregulated the expression of cell apoptosis pathway genes such as Cytochrome C, Fas, FasL, and Chop, both at the mRNA and at the protein levels. It also upregulated the expression of PAPP-A, IGF system, and VEGF, and so forth, which play important roles in the selection and dominance of growth follicles. The opposite results were observed by adding BMP1 antibody to the investigation groups. This study suggests that BMP1 regulates the proliferation and apoptosis of IVC GCs by changing the expression pattern of related genes and may potentially promote the selection and dominance of the buffalo follicles.