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Xiu Liu Zhongyan Shan Weiping Teng Department of Endocrinology and Metabolism,

Xiu Liu Zhongyan Shan Weiping Teng Department of Endocrinology and Metabolism, First Affiliated Hospital of China Medical University. Effect of MiR-125b-5p on Thyroid Hormon es Depletion Cultured C17.2 Neural Stem Cell s. Background. Thyroid hormones and brain development.

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Xiu Liu Zhongyan Shan Weiping Teng Department of Endocrinology and Metabolism,

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  1. Xiu Liu Zhongyan Shan Weiping Teng Department of Endocrinology and Metabolism, First Affiliated Hospital of China Medical University Effect of MiR-125b-5p on Thyroid Hormones Depletion Cultured C17.2 Neural Stem Cells

  2. Background Thyroid hormones and brain development • Thyroid hormones (THs) are essential for fetal neural development • THs regulate the neuronal cytoarchitecture,neuronal growth and synaptogenesis • The sensitive to the THs is not only remarked in the neonatal period but also prior to birth . Ahmed OM. Int. J. Devl Neuroscience 26 (2008) 147–209

  3. Neural inductio& Patterning Axon Extenison& Synapse Formation Differentiation Migration Proliferation Mir-9 Mir-134 Mir-9 Mir-25 Mir-124 Mir-34a Mir-128 Mir-273 Let -7 Mir-9 Mir-430 Mir-29a/b Mir-132 Mir-134 Background MicroRNA effects on brain development

  4. Background Our previous work MicroRNA expression profiling of rat offspring brains associated with maternal hypothyroidism and subclinical hypothyroidism during pregnancy in embryonic day 13 MiR-125b-5p

  5. Background Method used to study thyroid hormones deficency on cell lines • Depletion of L-3,5,3'-Triiodothyronine and L-Thyroxine in Euthyroid Calf Serum for Use in Cell Culture Studies of theAction of Thyroid Hormone • Regulation of Microglial Development: A Novel Role for ThyroidHormone • Effect of thyroid hormone depletion on cultured murine cerebral cortexastrocytes Endocrinology 1979,105: 80 The Journal of Neuroscience, 2001, 21(6):2028–2038 Neuroscience Letters 2009,467:58–62

  6. Objective • To identify the target gene of miR-125b-5p • To investigate the effect of miR-125b-5p on thyroidhormones depletion cultured C17.2 neural stem cells

  7. Materials and Methods mutant1 mutant2 http://www.targetscan.org/

  8. Materials and Methods Relative dual-Luciferase activity STAT3 3'UTR wt/mut-1/mut-2 Vectors and miR-125b-5p mimics co- transfected into HEK293 cells

  9. 50nM miR-125b-5p mimics 100nM miR-125b-5p mimics 100nM negative control 50nM negative control differentiationReal Time-PCR:Tubb3,Gfap target gene expression WB:stat3,p-stat3 proliferationCell Counting Kit-8 normal cultured thyroid hormone depletion cultured Materials and Methods C17.2 neural stem cells

  10. Results C17.2 neural stem cells nestin antibody and nuclei immune fluorescent staining

  11. Results Dual-luciferase reporter system assay * * *p<0.05

  12. Results Cell Counting Kit-8 proliferation assay * * *p<0.05

  13. Results Tubb3 and Gfap mRNA expression * * *p<0.05

  14. Results Total stat3 and phosphorylate stat3 protein expression normal cultured THs depletion cultured * * *p<0.05

  15. Conclusion • STAT3 is one of miR-125b-5p target genes • MiR-125b-5p may promote the differentiation of thyroid hormone depletion cultured C17.2 cell

  16. Thank You!

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