The Hypothalamus Sonic Hedgehog Positive Cell Lineage Tracing Analysis

Objective: SBE2(SBE2)is a specific enhancer of Shh in the hypothalamus region.SBE2-positive cells can specifically represent the distribution of Shh-expressing cells in the hypothalamus compared with the expression of endogenous Shh.SBE2 positive cells,and then can track the expression of Sonic hedgehog(Shh)positive cells in the hypothalamus and the developmental orientation of the progeny cells.At the same time,SBE2-Cre transgenic mice were constructed and tested whether they could be used as an important tool for studying the function of genes in the hypothalamus region.Methods: SBE2-Cre transgenic mice were constructed and mated by Rosa26-m Tm G and SBE2-Cre transgenic mice to obtain mouse embryos of E10.5,E12.5,E16.5 and p10 mice The Rosa26-m Tm G / SBE2-Cre double-positive mouse embryos were obtained by genotyping.The embryos were treated by PFA fixation,sucrose deposition and OCT embedding.A series of steps were taken,sliced and observed under fluorescence microscope.Period of SBE2 positive expression of regional changes in the location of the situation.Rosa26-Lac Z reporter gene mice were then mated with SBE2-Cre transgenic mice to obtain E10.5,E12.5 Rosa26-Lac Z / SBE2-Cre double positive embryos.The distribution of SBE2-positive cells in the brain was compared by Lac Z staining of embryos in each period.To demonstrate the above results using probe extraction,embryo whole in situ hybridization,in situ hybridization.And to obtain the expression position of Shh-positive cells in the brain of mouse embryos.It was further proved that the results of lineage tracking of SBE2 + cells were carried out.Results: SBE2 is a highly conserved sequence that acts as a Shh-specific enhancer to drive the expression of Shh in the hypothalamus and plays a key role in regulating the expression of Shh in the hypothalamus.In this study,SBE2-Cre mice were constructed and the Shh-positive cells were followed up by Rosa26-m Tm G and Rosa26-Lac Z reporter mice.The expression pattern of SBE2 in the hypothalamus of mice after embryonic development was observed.SBE2 was expressed in the preoptic area(POA)of mouse embryonic brain,Area,the entire expression area contains all kinds of neurons in the hypothalamus,in the ventral hypothalamus has a strong expression.The expression of SBE2 in the hypothalamus is a dynamic process,and the intensity of expression changes with the position changes.The expression of SBE2 was slightly different during the whole developmental stage from mouse embryos to birth,but in general it was a dynamic and continuous process.The expression of Shh in the brain of embryonic brain of E10.5 and E12.5 mice was compared with that of SBE2 line,and the results of SBE2 tracing were strongly supported.Conclusions: SBE2 + cells specifically represent the distribution of hypothalamic Shh-positive cells as one of the major cells for the differentiation of the hypothalamic nucleus.A series of experiments revealed that the hypothalamic Sonic hedgehog(Shh)positive cells and their Progeny cell development.After the previous study discussed in the hypothalamic Shh expression by a variety of transcription factors common regulation,SBE2 at the same time in all aspects of Shh regulation.And SBE2-Cre mice can be used as an important tool for studying the function of genes in the hypothalamus region,and this neurotrophic research plays an important role.