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Sonic Hedgehog Ectopic Expression Affects the Structure and Protein Expression of the Spinal Cord During Chicken Early Embryonic Development
Li Xiaoying1, Yang Ciqing1,2*, Wang Congrui2, Fu Sulei1, Li Han1, Lin Juntang1,2
1College of Life Science and Technology, Xinxiang Medical University, Xinxiang 453003, China; 2Key Laboratory of Henan Province for Medical Tissue Regeneration, Xinxiang 453003, China
Abstract: This research mainly analyzes the effect of Sonic Hedgehog (Shh) on the formation of the morphological structure of spinal cord and expression of related proteins during the chicken embryonic development. Using in ovo electroporation， 2 μg/μL pCAGGS-Shh and 0.25 μg/μL pCAGGS-GFP plasmid were mixed by 1:8 and then with 0.1~0.5 μL accurately injected into the spinal cord on E3 embryo. The condition of in ovo electroporation was voltage 18 V， each pulse 60 ms， interval 100 ms and total 6 times pulses. The positive embryos were collected after electroporation from 6 h to 5 d， for frozen sections. Fluorescent immunohistochemistry and DAPI staining were carried out to observe the change of the morphological structure and the related proteins expression. The results show that GFP expression was observed in 6 h after in ovo electroporation. After 24 h of electroporation， it was detected that the Shh ectopic expression induces the expression of Nkx2.2 (NK2 homeobox 2) in the spinal cord， however Pax7 (paired-type homeobox) expression was inhibited， furthermore， Shh ectopic expression leads to a change of the spinal cord structure. As an important signaling molecule， ectopic expression of Shh induces or inhibits the expression of related proteins， and leads to the spinal cord malformations.
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