Furuta A, Rothstein J D, Martin L J
Department of Neurology, Division of Neuropathology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287, USA.
J Neurosci. 1997 Nov 1;17(21):8363-75. doi: 10.1523/JNEUROSCI.17-21-08363.1997.
Extracellular glutamate concentrations are regulated by glial and neuronal transporter proteins. Four glutamate transporter subtypes have been identified in rat brain; GLAST and GLT-1 are primarily astrocytic, whereas EAAC1 and EAAT4 are neuronal. Using immunoblotting and immunohistochemistry with subtype-specific antipeptide antibodies, we examined the protein expression and regional and cellular localization of each glutamate transporter subtype in embryonic and postnatal rat CNS. Each transporter had a specific pattern of expression. GLAST immunoreactivity was low prenatally but became enriched in cerebellar Bergmann glia early postnatally and then was also present in forebrain later postnatally. The post-translational modification of GLAST was unique among the subtypes; glycosylated GLAST increased with maturation, whereas nonglycosylated protein decreased in abundance postnatally. GLT-1 was present in fetal brain and spinal cord, with expression progressively increasing to adult levels throughout the neuraxis by postnatal day 26. Transient expression of GLT-1 immunoreactivity along axonal pathways was observed prenatally, in contrast to the exclusive localization of GLT-1 to astrocytes in the adult CNS. EAAC1, localized to neurons, was enriched in forebrain, diencephalon, and hindbrain during prenatal and postnatal development. EAAC1 expression was greater in newborn brain compared with adult brain. EAAT4 had a region-specific distribution; EAAT4 was mainly in cerebellum, localized to Purkinje cells, with much lower levels in forebrain. EAAT4 levels increased in cerebellum with age. We conclude that during CNS development the expression of glutamate transporter subtypes is differentially regulated, regionally segregated, and coordinated.
细胞外谷氨酸浓度由胶质细胞和神经元转运蛋白调节。在大鼠脑中已鉴定出四种谷氨酸转运体亚型;谷氨酸天冬氨酸转运体(GLAST)和谷氨酸转运体1(GLT-1)主要存在于星形胶质细胞中,而兴奋性氨基酸载体1(EAAC1)和兴奋性氨基酸转运体4(EAAT4)存在于神经元中。我们使用免疫印迹法和针对亚型特异性抗肽抗体的免疫组织化学方法,研究了胚胎期和出生后大鼠中枢神经系统中每种谷氨酸转运体亚型的蛋白表达、区域定位和细胞定位。每种转运体都有特定的表达模式。GLAST免疫反应性在出生前较低,但在出生后早期在小脑伯格曼胶质细胞中富集,随后在出生后晚期也出现在前脑中。GLAST的翻译后修饰在各亚型中是独特的;糖基化的GLAST随着成熟而增加,而非糖基化蛋白在出生后丰度降低。GLT-1存在于胎儿脑和脊髓中,到出生后第26天,其表达在整个神经轴中逐渐增加至成年水平。与成年中枢神经系统中GLT-1仅定位于星形胶质细胞相反,在出生前观察到GLT-1免疫反应性沿轴突途径短暂表达。定位于神经元的EAAC1在产前和产后发育期间在前脑、间脑和后脑富集。与成年脑相比,新生脑中EAAC1的表达更高。EAAT4具有区域特异性分布;EAAT4主要位于小脑,定位于浦肯野细胞,在前脑中水平低得多。随着年龄增长,小脑中EAAT4水平升高。我们得出结论,在中枢神经系统发育过程中,谷氨酸转运体亚型的表达受到差异调节、区域分隔和协调。