ESPE Abstracts (2014) 82 P-D-3-1-810

Growth

The Role of IGF1R Gene Mutation in the Development of Oligodendrocytes

Fan Yang, Mei Tang & Hedong Li

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West China 2nd University Hospital, Sichuan University, Chengdu, Sichuan, China


Background: IGF1R gene mutation usually cause IUGR. The children born with IUGR were prone to some kinds of brain function disorders.

Objective and hypotheses: The dysfunction of the brain was caused by the abnormal oligodendrocyte development.To establish lentivirus vector of IGF1R gene mutation (R709Q) and transfect oligodendrocyte precursors (Ge6). Observe the IRS/MAPK and PI3K/Akt/PKB signaling pathway and the change of proliferation, differentiation, and apoptosis of oligodendrocyte.

Method: Synthesise IGF1R (R709Q) gene in vitro and clone in lentivirus vector with resistance to puromycin (puro). R709Q gene was mediated by lentivirus and to transfect Ge6 cells. Target cells were selected by puro. The experimental group R709Q cells and the control group Ge6 cells were culture in vitro. IGF1 were used respectively. Immunofluorescent staining were applied to observe the positive rate of caspase-3 and O4; western blot were applied to observe the IRS/MAPK and PI3K/Akt/PKB signaling pathway, which is the proportional change of p-Erk1/Erk2, p-Akt, p-Bad and Erk1/Erk2, Akt, Bad.

Results: Immunofluorescent staining indicated that the caspase-3 positive rate of experimental group increased compared to the control group (P<0.05). The other Immunofluorescent staining indicate that the O4 positive rate of experimental group decreased compared to the control group (P<0.05); Western blot indicated that p-Erk1/Erk2/ Erk1/Erk2 Protein expression levels of experimental group are higher than that control group (P<0.05), however P-Akt/Akt, p-Bad/Bad protein expression levels of experimental group are lower in control group (P<0.05).

Conclusion: The expression of IRS/MAPK and PI3K/Akt/PKB signaling pathway is changed in mutated oligodendrocytes which lead to abnormal proliferation, differentiation and apoptosis.

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