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培养的分离小鼠少突胶质细胞对γ-[³H]氨基丁酸的高亲和力摄取。

High-affinity uptake of gamma-[3H]aminobutyric acid by isolated mouse oligodendrocytes in culture.

作者信息

Reynolds R, Steffen C, Herschkowitz N

机构信息

Department of Pediatrics, University of Berne, Inselspital, Switzerland.

出版信息

Neurochem Res. 1987 Oct;12(10):885-90. doi: 10.1007/BF00966310.

Abstract

Oligodendrocytes were isolated from mixed glial cultures of neonatal mouse forebrain and further grown in serum-free hormone supplemented culture medium. Cell populations were identified by indirect immunofluorescence using a range of specific antibodies, revealing a predominantly immature population of oligodendrocytes, the majority expressing the myelin glycolipids galactocerebroside and sulfatide on their plasma membrane. Astroglial contamination was found to be minimal. Simultaneous autoradiography and immunofluorescence demonstrated the presence of a transport system for the major inhibitory neurotransmitter GABA in the oligodendrocytes. The transport system was found to be energy, sodium and temperature dependent. Kinetic analysis revealed a high affinity system, with a Km of 6.27 microM and Vmax of 0.714 nmol/min/mg protein, which is comparable to that found previously for CNS neurons and astrocytes.

摘要

少突胶质细胞从小鼠新生前脑的混合胶质细胞培养物中分离出来,并在添加了激素的无血清培养基中进一步培养。使用一系列特异性抗体通过间接免疫荧光鉴定细胞群体,结果显示主要是未成熟的少突胶质细胞群体,大多数细胞在其质膜上表达髓鞘糖脂半乳糖脑苷脂和硫脂。发现星形胶质细胞污染极少。同时进行的放射自显影和免疫荧光显示少突胶质细胞中存在主要抑制性神经递质γ-氨基丁酸(GABA)的转运系统。发现该转运系统依赖能量、钠和温度。动力学分析揭示了一个高亲和力系统,其米氏常数(Km)为6.27微摩尔,最大反应速度(Vmax)为0.714纳摩尔/分钟/毫克蛋白质,这与先前在中枢神经系统神经元和星形胶质细胞中发现的情况相当。

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