Analysis Of The Whole Genome Expression Of CMT2L Transgenic Mice Using Gene Chip Technology

Objective:To study and compare the whole genome expression in sciatic nerve in CMT2L transgenic mice, wtHSPB8transgenic mice and B57CL mice by using MouseRef-8v2.0Expression BeadChip (Illumina). We filter genes having altered expression by biological informatics analysis, and verified their altered expression by fluorescent quantitative (RT-PCR) in the sciatic nerve. And we clarify the changes of genetic expression in mutation of wtHSPB8in different period of CMT2L, find out the possible passageway of intrinsic molecular, and discuss probable pathogenesis of mutation of wtHSPB8in CMT2L.Methods:(1)CMT2L transgenic mice and wtHSPB8transgenic mice has already been created and verified successfully. Being the object of study, the transgenic mice ranging from3months of age, six months of age and18months are compared with the matching transgenic mice of wtHSPB8transgenic mice and B57CL mice.(2)Differential expression genes are filtered by using whole genome expression BeadChip (Illumina). We extract mRNA in sciatic nerve of mice and analyze the differential gene expression in CMT2L transgenic mice, wtHSPB8transgenic mice and B57CL mice by using MouseRef-8v2.0Expression BeadChip (Illumina).(3)We verify altered expression by fluorescent quantitative (RT-PCR) in the sciatic nerve. Four CMT2virulence genes and genes having altered expression have been chosen and their differential gene expressions in the sciatic nerve are tested by fluorescent quantitative (RT-PCR).Results:(1)51differential expression genes in sciatic nerve have been found out by biological informatics analysis. In which31genes are up-regulated and20are down-regulated.Next, we cluster51differential expression genes according to protein-related in this experiment. It shows that8genes are related to muscle protein and6are related to motor protein binding. Then, we group51differential expression genes according to their unique function and find that7of them are relevant to muscle contraction. Besides,5synthetic genes in relation to carbohydrate (fbp2, pgam2, rbp4, pckl, ppplr3c) are discovered through gene ontology analysis, the rest of genes being with no related polymerization as well as special meanings.(2) We verify altered expression by fluorescent quantitative (RT-PCR) in the sciatic nerve, group51altered expression genes according to biological function and analyze the passageway of intrinsic molecular.7altered expression genes named fbp2, pgam2, rbp4, pckl, ppp1r3c, myh4and neb have been verified by fluorescent quantitative (RT-PCR). Differential genes relating to CMT2have not been found in gene chip in this experiment. We choose11virulence genes relating to CMT2and verified them by fluorescent quantitative (RT-PCR). it shows that genes called fbp2,pgam2,rbp4,pckl,ppp1r3c all rise up, which are agree with the result of chip. However genes named myh4and neb2have not waved much in genetic expression. While virulence genes, mfn2, rab7, nefl and hspbl, change little in genetic expression.Conclusion.Through the analysis of whole genome chip and the verification of fluorescent quantitative (RT-PCR), we find Genes which are synthesized with carbohydrate--fbp2,pgam2,rbp4,pckl and ppplr3c all rise up in CMT2L transgenic mice, suggesting that there may be a connection between the morbidity of CMT2L and the abnormal decrease of carbohydrate metabolism.