The Expression Of Mxra7 Gene In Ontogeny And Tissues

Objective Matrix remodelling associated 7(MXRA7)is a kind of gene that predicated by bioinformatics analysis.At present,there is no any report about expressional and functional study of the gene.In previous studies,we applied the expression microarray of the whole genome and quantitative Polymerase Chain Reaction to find the gene presenting a time-dependent decrease and recovery during the models of corneal neogenesis pannus and infection,while other genes(eg.MMP3 and MMP13)associated with corneal damage show an adverse trend.The expression of the gene in the 14.5d mouse embryo was specially discovered by hybridization in situ in the retinal,corneal epithelium and lens epithelial cells while other tissues was not.We applied the technology of gene knockout to study the effect on development of eyes in mice;Preparing monoclonal antibodies to consider the expression of this gene at different stages;Various kinds of pathological or diseased models were builded to observe the changes of the gene.By building various animal models to find the phenotype that influenced the distribution of Mxra7,in order to study the role of the gene in liver injury and in tumor angiogenesis.Methods We introduced three pairs of heterozygous mice which were knocked out of Mxra7(Mxra7+/-)to carry on amplifyng..Mouse tails were cut to be extracted deoxyribonucleic acid(DNA)to identify the type.Wild-type(WT)mice and homozygote mice(Mxra7-/-)were mated separately to reproduce.Monoclonal antibody that combined with predictive protein of this gene was established.WT mice and homozygote mice were killed and then the hearts,livers,lungs,kidneys,brains and eyeballs were taken down to be fixed with 4%paraformaldehyde and opti-mum cutting temperature compound(OCT).Various kinds of tissues were stored at-80℃freezer to extract total protein.We adopted immunofluorescence and immunohistochemistry to dye monoclonal antibody to detect the expressioned sites and dynamic changes of MXRA7 protein,and applied western blot to discover the amount of this protein in different tissues.Human tumor tissue sections were used to comapre with mouse in expression of the protein.By building various animal models to find the phenotype that influenced the distribution of MXRA7,for example,hepatic injury model or matrigel model were injected CCL4 or matrigel glue via abdominal cavity.Results The mated breeding mice were determined the type through DNA identification.Our study indicated MXRA7 protein was distributed in heart,liver,lung,kidney and brain,and was the highest expression in liver,mainly concentrated around the blood vessel walls in the hepatic injury sections by immunofluorescence and immunohistochemistry.The MXRA7 protein concentration is higher in heart,liver,lung and kidney in wild mice than homozygote mice.The amount of blood platelets of wild mice recovered soon than homozygote mice in hemopoiesis injury model.Neovascularization was less distributed in knockout mice comparing with wild type in matrigel model.The weights and sizes of tumors of homozygote mice are larger than wild mice in mouse models of medulloblastoma.