Expression of SHH and IHH during the formation of intervertebral discs in mouse embryo / 中国组织工程研究

ABSTRACT

BACKGROUND: Studies have shown that Sonic Hedgehog (Shh) is expressed in mouse Notochordal cells, and India Hedgehog (Ihh) is mainly expressed in chondrocyte-like cells and end plates of mouse embryonic vertebral bodies. However, the expression of Shh and Ihh during the formation of intervertebral discs in mouse embryo is unclear. OBJECTIVE: To investigate the expression of Shh and Ihh during the formation of intervertebral discs in mouse embryo. METHODS: Male Shh-CreERT2, R26-mTmG/+ mice mated with female R26-mTmG/+ mice to obtain pregnant rats at different periods of pregnancy (E8.5, E11.5, E12.5, E14.5, E16.5, E18.5), followed by injection of 10 g/L tamoxifen at a dose of 10 μL/g. Intervertebral disc tissues were isolated from the mouse embryo at P0, followed by genotype identification. At the same time, male C57BL/6 mice mated with female C57BL/6 mice to obtain pregnant rats at different periods of pregnancy (E11.5, E12.5, E14.5, E16.5, E18.5), and the intervertebral disc tissues of mouse embryo were taken and analyzed. The expression of Shh and Ihh during the formation of intervertebral discs in mouse embryo was detected using immunofluorescence and immunohistochemistry, respectively. Implementation of animal experiments was approved by the Animal Ethics Committee of Soochow University. RESULTS AND CONCLUSION: Shh-CreERT2; R26-mTmG/+ mouse embryos were identified by PCR amplification. The results of immunofluorescent staining showed a gradual decrease in the expression of Shh in nucleus pulposus cells during the formation of intervertebral discs in mouse embryo. The results of immunohistochemical staining showed a gradual increase in expression of Ihh in nucleus pulposus cells during the formation of intervertebral discs in mouse embryo. In summary, Shh and Ihh are dynamically expressed during the formation of intervertebral discs in mouse embryo, and provide a basis for further research on the molecular mechanism of intervertebral disc degeneration.