Analysis Of Neuritin Expression And The Proteins Interacting With It

Neuritin was a novel neurotrophic factor which was discovered from the adult rat hippocampus cDNA library by Israel scientist Nedivi in 1993. The expression product of neuritin can enhance the development of neuron axon arbors by promoting synaptogenesis and by increasing the addition of new axon branches, and its expression was induced by neural activity or neurotrophins, which indicated that the neural activities or neurotrophins function were related to neuritin.Objective: The aim of project is to identify the tissue expression profile and intracellular localization of neuritin, and to find proteins that interact with Neuritin using yeast two hybrid system, which provide theoretical basis for further research about its function and mechanism.Method: neuritin structure and its potential function and Serial Analysis of Gene Expression(SAGE) were analyzed using bioinformation technology; neuritin ORF was amplified from human cDNA library, and then neuritin expression profile was showed with RT-PCR and immunohistochemistry; Intracellular localization of neuritin was inferred by immunohistochemistry and eukaryon expression vector combinded with green fluorescence protein, pEGFP-C1-neuritin; Screened a human fetal brain cDNA library with neuritin cDNA as bait using Gal4 yeast two hybrid system.Results:1) Analysis of neuritin structure, expression profile and its potential function with bioinformation technology. 2)neuritin expression profile:multiple Tissure cDNA panel and immunohistochemistry showed that Neuritin expression in a high level in many human normal tissues, especially in ganglion cell of brain and liver. 3) neuritin intracellular location: the result from green fluorescence assay showed Neuritin location in cytoplasm, which was consistent with result of immunohistochemistry. 4)Using yeast two hybrid, screened a human fetal brain cDNA library with neuritin cDNA as bait, and found that the proteins coded by 59 genes had interaction with Neuritin.Conclusion: 1) MTC Panel, multiple tissue immunohistochemistry and SAGE showed that Neuritin was expressed not only in brain, but in heart, liver, lung,intestine and ovary. The results implied that Neuritin play a critical role beyond in nerval system. 2) The results from green fluorescence protein orientation and immunohistochemistry showed that Neuritin was located in cytoplasm. 3. yeast two hybrid results showed that Neuritin interacted with the proteins related to nerve, tumor and signal transduction respectively. These data indicated that Neuritin was possibly relative to cell proliferation and tumorigenesis, as well as to nerves plasticity.