关键词: 胚胎, 绿色荧光蛋白, α-平滑肌肌动蛋白, 神经丝蛋白, 活体电转, 鸡

Abstract: P>Objective The method of in vivo electroporation has been set up successfully and we further analyzed the effect of report gene green fluorescence protein(GFP) on the morphology of developing chicken embryos after in vivo electroporation, and also analyzed the expression of α-smooth muscle actin (α-SMA) and neurofilament during chicken embryonic development. Methods pCAGGS-GFP was transformed into chicken embryos with in ovo culture at 3days and ex ovo culture at 3-5days. In 24hours after in vivo electroporation, GFP-positive embryos were selected under stereo fluorescence microscope, and the GFP-negative embryos served as controls. Five embryos were analayzed for each group. Fluorescence immunohistochemistry was applied to analyze the expression ofα-SMA and neurofilament in chicken spinal cord and tectum. Results At different stages of chicken embryos and different time after in vivo electroporation, the expression of α-smooth muscle actin and neurofilament did not show difference in experimental group and wild type, as well as in GFP-positive area and GFP-negative area. The morphology of embryos was not changed after electroporation with pCAGGS-GFP either. BR>Conclusion GFP as a report gene to in vivo electroporation for chicken embryos does not affect the expression of α-smooth muscle actin and neurofilament, as well as no effect on the morphology of chicken embryos, so GFP

Key words: Embryo, Green fluorescence protein, α-smooth muscle actin, Neurofilament, EM>in vivo/EM> electroporation, Chicken