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Influence of sonic hedgehog overexpression on Pax family in the spinal cord during chicken embryonic development
LI Han YANG Ci-qing QIAO Liang ZHANG Bi-chao LIN Jun-tang*
Acta Anatomica Sinica ›› 2015, Vol. 46 ›› Issue (5) : 667-672.
Influence of sonic hedgehog overexpression on Pax family in the spinal cord during chicken embryonic development
Objective To study the effect of sonic hedgehog (Shh) gene overexpression on Pax family in the spinal cord during chicken embryonic development.
Methods At E2.5-E3, Shh expression plasmids and green fluorescence protein (GFP) reporter gene plasmids were co-transfected into chicken embryonic spinal cord by in vivo electroporation. Only GFP reporter gene plasmids were transfected by the same way as control. Samples were collected from three or more embryos from the experimental and control groups and sliced at E4. Thenin situhybridization was used to detect whether Shh was really overexpressed. Immunofluorescence histochemistry was used to study whether Pax3, Pax6 and Pax7 were regulated by the overexpressed Shh.Statistical software was utilized to analyze the expression quantity of Pax3, Pax6 and Pax7 between two sides of the chicken spinal cords in the control and experimental group. Results The results ofin situhybridization showed that Shh was mainly expressed in the floor plate and notochord and the overexpression model of Shh in chicken spinal cord was built successfully in the experimental group. The immunohistochemical positive staining for Pax3 was distributed in alar plate,dermatome, dorsal root and dorsal root ganglion in the E4 chicken embryo spinal cord. The immunofluorescence positive signal for Pax6 was in the middle of spinal cord across the alar plate and the basal plate. The signal of Pax7 mainly existed evenly in the alar plate roof plate and dermatome. There was highly significant difference between electroplated side and normal side (P<0.01). The expressions of Pax3, Pax6 and Pax7 were all depressed by Shh overexpression. Conclusion There are different expression patterns in Pax3,Pax6,and Pax7 in the developmental chicken spinal cord, and the intensities of immunofluorescence positive signal for these genes are decreased by Shh overexpression, which indicates that they are regulated by Shh signal pathway.
Sonic hedgehog /
Paired box /
Chicken spinal cord in vivo electroporation /
In situ hybridization /
Chicken
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