• BMP10 is a secreted protein and after binding to its receptors, SMAD family transcription factors were recruited and activated, leading to gene expression.
• The exons 1-2 of mouse Bmp10 gene that encode the whole molecule, were replaced by human counterparts in B-hBMP10 mice. The promoter, 5’UTR and 3’UTR region of the mouse gene are retained. The human BMP10 expression is driven by endogenous mouse Bmp10 promoter, while mouse Bmp10 gene transcription and translation will be disrupted.
• Mouse and human BMP10 were detectable in plasma, serum and heart both from wild-type and homozygous mice, as the antibodies used showed cross-recognition for mouse and human BMP10.
• Homozygous mice expressed the human BMP10 and developed normally.
• This model is useful in studying the role of BMP10 in liver cancer and heart development.

Gene targeting strategy for B-hBMP10 mice. The exons 1-2 of mouse Bmp10 gene that encode the whole molecule, were replaced by human counterparts in B-hBMP10 mice. The promoter, 5’UTR and 3’UTR region of the mouse gene are retained. The human BMP10 expression is driven by endogenous mouse Bmp10 promoter, while mouse Bmp10 gene transcription and translation will be disrupted.

Strain specific analysis of BMP10 mRNA expression in wild-type C57BL/6N mice and B-hBMP10 mice by RT-PCR. Heart RNA were isolated from wild-type C57BL/6N mice (+/+) and homozygous B-hBMP10 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human BMP10 primers. Mouse Bmp10 mRNA was detectable only in wild-type C57BL/6N mice. Human BMP10 mRNA was detectable in homozygous B-hBMP10 mice, but not in wild-type C57BL/6N mice.

Western blot analysis of BMP10 protein expression in homozygous B-hBMP10 mice. Plasma and serum were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hBMP10 mice (H/H), (n=2, 11-week-old male mice) and then analyzed by western blot with anti-BMP10 antibody (R&D, MAB2926). 40 μg total proteins were loaded for western blotting analysis. The antibody used showed cross-recognition for mouse and human BMP10.

Western blot analysis of BMP10 protein expression in homozygous B-hBMP10 mice. Heart, liver, spleen and kidney were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hBMP10 mice (H/H), (11-week-old male mice) and then analyzed by western blot with anti-BMP10 antibody (R&D, AF3956). 40 μg total proteins were loaded for western blotting analysis. BMP10 was only detectable in heart both from wild-type and homozygous mice, as the antibody used showed cross-recognition for mouse and human BMP10.