alterations of gene expression and glutamate clearance in astrocytes derived from an mecp2-null mouse model of rett syndrome改变基因表达和谷氨酸的间隙在星形胶质细胞来源于一个mecp2-null rett综合症的小鼠模型.pdfVIP

Alterations of Gene Expression and Glutamate Clearance in Astrocytes Derived from an MeCP2-Null Mouse Model of Rett Syndrome Yasunori Okabe1,2, Tomoyuki Takahashi1,3*, Chiaki Mitsumasu1,3, Ken-ichiro Kosai1,3,4, Eiichiro Tanaka1,2, Toyojiro Matsuishi 1,3 1 Division of Gene Therapy and Regenerative Medicine, Cognitive and Molecular Research Institute of Brain Diseases, Kurume University, Kurume, Japan, 2 Department of Physiology, Kurume University of Medicine, Kurume, Japan, 3 Department of Pediatrics, Kurume University of Medicine, Kurume, Japan, 4 Department of Gene Therapy and Regenerative Medicine, Advanced Therapeutics Course, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan Abstract Rett syndrome (RTT) is a neurodevelopmetal disorder associated with mutations in the methyl-CpG–binding protein 2 (MeCP2) gene. MeCP2-deficient mice recapitulate the neurological degeneration observed in RTT patients. Recent studies indicated a role of not only neurons but also glial cells in neuronal dysfunction in RTT. We cultured astrocytes from MeCP2- null mouse brain and examined astroglial gene expression, growth rate, cytotoxic effects, and glutamate (Glu) clearance. Semi-quantitative RT-PCR analysis revealed that expression of astroglial marker genes, including GFAP and S100b, was significantly higher in MeCP2-null astrocytes than in control astrocytes. Loss of MeCP2 did not affect astroglial cell morphology, growth, or cytotoxic effects, but did alter Glu clearance in astrocytes. When high extracellular Glu was added to the astrocyte cultures and incubated, a time-dependent decrease of extracellular Glu concentration occurred due to Glu clearance by astrocytes. Although the shapes of the profiles of Glu concentration versus time for each strain of astrocytes were grossly similar, Glu concentration in the m