RESUMO
This study was purpose to establish the transgenic mouse models of the truncated platelet integrin ß3 by retrovirus-infected hematopoietic stem cells (HSCs) transplantation and to provide the basis for further study of the role of integrin ß3 cytoplasmic domain in platelet bi-directional signaling pathways. Wild-type ß3, ß3-Δ759 (R(760) GT(762) truncated ß3) and ß3-Δ754 (T(755) NITYRGT(762) truncated ß3) cDNAs were subcloned into MSCV MigR1 retroviral vector bearing a GFP gene and packaged into infective retrovirus with BOSC23 cell strain. The bone marrow HSCs of the ß3 deficient mice were infected by the retroviruses, and transplanted into lethally-irradiated wild type C57BL/6 mice. GFP positive rate and surface ß3 expression of the recipients' platelets at 6 to 8 weeks after transplantation were detected by flow cytometry to evaluate the transgenic efficiency. The results showed that four kinds of transgenic mouse models including vector, wild-type ß3, ß3-Δ759 and ß3-Δ754 were established successfully. GFP positive rates of transgenic mouse platelets ranged from 18% to 66% and the ß3 expression of transgenic mouse reached heterozygote (ß3(+/-) level of mouse). It is concluded that establishment of transgenic mouse models mediated by retrovirus-infected HSCs transplantation is a feasible, fast, and high throughput transgenic approach and laid a solid foundation for further research on the role of integrin ß3 cytoplasmic domain for bi-directional signaling of platelets in vivo, and for the gene therapy of platelet disorders.
