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The molecular cloning of glial fibrillary acidic protein in Gekko japonicus and its expression changes after spinal cord transection

机译:壁虎脊髓横断后胶质纤维酸性蛋白的分子克隆及其表达变化。

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摘要

The glial fibrillary acidic protein (GFAP) is an astrocyte-specific member of the class III intermediate filament proteins. It is generally used as a specific marker of astrocytes in the central nervous system (CNS). We isolated a GFAP cDNA from the brain and spinal cord cDNA library of Gekko japonicus, and prepared polyclonal antibodies against gecko GFAP to provide useful tools for further immunochemistry studies. Both the real-time quantitative PCR and western blot results revealed that the expression of GFAP in the spinal cord after transection increased, reaching its maximum level after 3 days, and then gradually decreased over the rest of the 2 weeks of the experiment. Immunohistochemical analyses demonstrated that the increase in GFAP-positive labeling was restricted to the white matter rather than the gray matter. In particular, a slight increase in the number of GFAP positive star-shaped astrocytes was detected in the ventral and lateral regions of the white matter. Our results indicate that reactive astrogliosis in the gecko spinal cord took place primarily in the white matter during a short time interval, suggesting that the specific astrogliosis evaluated by GFAP expression might be advantageous in spinal cord regeneration.
机译:胶质纤维酸性蛋白(GFAP)是III类中间丝蛋白的星形胶质细胞特异性成员。它通常用作中枢神经系统(CNS)中星形胶质细胞的特异性标记。我们从日本壁虎的大脑和脊髓cDNA文库中分离出GFAP cDNA,并制备了针对壁虎GFAP的多克隆抗体,为进一步的免疫化学研究提供了有用的工具。实时定量PCR和Western印迹结果均显示,横断后脊髓中GFAP的表达增加,在3天后达到最高水平,然后在实验的其余2周逐渐减少。免疫组织化学分析表明,GFAP阳性标记的增加仅限于白质而不是灰质。特别地,在白质的腹侧和外侧区域中检测到GFAP阳性星形星形胶质细胞的数量略有增加。我们的结果表明,壁虎脊髓中的反应性星形胶质沉着症主要发生在短时间内的白质中,这表明通过GFAP表达评估的特定星形胶质沉着症可能对脊髓再生有利。

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