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PITX2 is a member of PITX family, with a paired-like homeodomain.PITX2 protein plays a crucial role in cell proliferation, differentiation and organogenesis through the Wnt/beta-catenin pathway.Meanwhile, it also involved in POU1F1 pathway, which is a well known pathway in lactation, growth and development in the mammalians.Alternative splicing plays an important role in regulating biological function and functional diversity in organisms.Thus, this study focused on detecting the isoforms of bovine PITX2 and their mRNA expression characterization.Herein, two novel isoforms, PITX2 and PITX2-V1, were identified in bovine by RT-PCR.The entire coding sequences of PITX2 and PITX2-V1 were 978 bp (encoding 325 amino acids) and 354bp (encoding 118 amino acids) in length, respectively.Bovine PITX2 has perfect homeodomain and OAR domain, but PITX2-V1 is lack of homeodomain.RT-PCR revealed that the transcripts of bovine PITX2 and PITX2-V1 expressed in most tissues.But PITX2 expressed in a very lower level in the testis, and PITX2-V1hardly expressed in the brain and testis.Interestingly, PITX2 and PITX2-V1 were both highly expressed in ovary,which indicted their function in regulating ovary development or milk production.All these findings would provide a foundation for the further research ofPITX2 gene in bovine growth, development and lactation.