The Mechanism Of Fos And Leptin Receptor In Neural Tube Development Of Offspring Under Different Energy Metabolism

Objective The aim is the correlation between fos and leptin-LEPR during the development of nerve tubes in different energy states and to study its effects on the development of nerve tubes in offspring.Methods1.Using the gene identification technology of our research group,we obtained 4-week Mex3c^+/-female mice as a negative energy state（Mex3c group,n=30）;in 4 weeks age control female mice（60）,half of all was fed a high-fat diet as a positive energy state（HFD group,n=30）;another was fed chow diet as a control group（control group,n=30）,sustained to 14 weeks.The E10.5d（n=10）,E12.5d（n=10）and E14.5d（n=10）were got by male and female mice（1:3）.2.The female mice,liver/abdominal fat/kidney,in various energy states were stained with HE method.liver and kidney cells was examined by IHC and WB to test leptin and LEPR.3.The shap of the neural tube was appered by HE staining and mortality was found by Tunel testing.In placenta,the mRNA content of leptin receptor（LEPR）and fos and in embryo the mRNA content of leptin,LEPR,nestin and fos were detected by RT-PCR.The expression of LEPR in nerve tubes was recognized by in situ hybridization.The expression of leptin,LEPR,nestin,FOS,Pax3,H3K27me3 proteins in the neural tube of the offspring was detected.Results1.At 14 weeks age,the three groups about body weights were tesed,in the control group（18.82g±1.54g）;in mex3c group（18.84g±1.08g）and in the HFD group（22.61g±1.10g）.2.Female mice metabolic status on various energy states:⑴Pathological staining results,compared with the Control group,the HFD group had large lipid droplets and decreased cytoplasm;there were lipid droplets in the liver cells of the Mex3c^+/-mutation group.⑵Pathological staining results showed that compared with the Control group,the renal tubular cells of the HFD group were vacuolated,the cells were disordered,and the glomeruli were shrunk;the continuity of the epithelial cells of the renal tubules in the Mex3c group was interrupted,and some cells showed vacuoles.⑶The results of leptin,LEPR protein:At the liver,the HFD group and the Mex3c group,the results of LEPR was shown low（P<0.05）were contrast with control group.In the kidney,the testing results of the leptin:Control group<Mex3c group<HFD group（P<0.01）,while the LEPR was totally different from that of leptin（P<0.01）.3.Development of neural tubes in different energy states（1）E10.5d,E12.5d,E14.5d embryo specimens were collected under different energy states,and the total number of embryos,live fetuses,and absorbed fetuses were not statistically significant,and the embryos under the stereo microscope were generally not different.（2）HE staining of the nerve tube:the maturity on nerve cells decreased in the Mex3c group and the HFD group was rather than the control group.（3）In the nerve tube of the offspring,the expression of fos,leptin and LEPR was discovered.Immunohistochemical stains indicated that fos and leptin were expressed in the nucleus.The cell membrane were found LEPR.WB showed that the Mex3c and HFD groups are low expressed in fos proteins（P<0.01）,the Mex3c group is highly expressed in the LEPR proteins（P<0.01）and the HFD group（P<0.05）,which respectivelywere contrast with control group.In the HFD group leptin hyper-expressions（P<0.05）.At different stages of development,there was different leptin expressions.In the E12.5d neural tubes,the HFD and Mex3c groups,the leptin protein is low（P<0.01）with contrast the control group.In the E14.5d neural tubes,the Mex3c group expresses the leptin protein highly（P<0.05）than control group,but the HFD group does not show very differences.4.Fos,leptin,LEPR and nestin mRNA by RT-PCR test results.In the placental tissue,there is a high expression about fos of Mex3c group（P<0.05）and there is a low fos of HFD group（P<0.05）,which all were contrast with other group.while in the placenta E14.5d,it in three groups do not differ from each other.In the neural tube,the expression about nestin,PAX3,H3K27me3,the results of the immunization experiment showed that nestin expresses the nerve fibers,PAX3 and H3K27me3 on the cell nuclei.The results of the Western Blooting experiment showed that the Mex3c groups highly produced the protein nestin（P<0.01）,PAX3（P<0.01）,H3K27me3（P<0.01）was contrast with control group;at the same time,the HFD group has similar to Mex3c group.Conclusion（1）The expression trends of leptin and LEPR in the kidney tissues of mother rats are oposite of different energy states.（2）Mex3c regulates leptin and LEPR by enhancing the expression of fos mRNA to participate in the neural tube development of the offspring.（3）The neural tube nestin,PAX3,and H3K27me3 of the offspring of the Mex3c group and the HFD group are consistently highly expressed,which preliminarily reveals that different energy states regulate leptin-LEPR to take part in the process of neurodevelopment.Positive and negative energy slows down the development of the nervous system.