Yamada K, Watanabe M, Shibata T, Nagashima M, Tanaka K, Inoue Y
Department of Anatomy, Hokkaido University School of Medicine, Sapporo 060-8638, Japan.
J Neurosci. 1998 Aug 1;18(15):5706-13. doi: 10.1523/JNEUROSCI.18-15-05706.1998.
The glutamate transporter GLT-1 is expressed in astrocytes of the mature brain and spinal cord. In the present study, we examined its expression in the developing mouse spinal cord. By in situ hybridization, 35S-labeled antisense oligonucleotide probes for GLT-1 mRNA consistently labeled the mantle zone/gray matter from embryonic day 11 through the adult stage. However, immunohistochemistry with a specific antibody visualized distinct regional and cellular localizations during the time between the fetal and postnatal stages. At fetal stages, GLT-1 immunoreactivity predominated in the marginal zone/white matter, observed as tiny puncta in cross-sections and as thin fibers in longitudinal sections. The GLT-1-immunopositive structures were also labeled for neuron-specific enolase, a glycolytic enzyme specific to postmitotic neurons and endocrine cells. By electron microscopy, GLT-1 immunoreactivity was detected in axons forming frequent enlargements and was focally localized on a small portion of the axolemma, particularly that facing adjacent axons. At early postnatal stages, GLT-1 disappeared from axons in white matter tracts and, instead, appeared in astrocytic processes surrounding various neuronal elements in the gray matter. Therefore, before switching to astrocytic expression, GLT-1 is transiently expressed in neurons and localized in differentiating axons. Together with our previous finding on the localization of glutamate transporter GLAST in radial glial fibers, GLT-1 and GLAST are thus localized during development on distinct directional cellular elements along which young neurons elongate their axons or move their cell bodies, respectively.
谷氨酸转运体GLT - 1在成熟脑和脊髓的星形胶质细胞中表达。在本研究中,我们检测了其在发育中的小鼠脊髓中的表达。通过原位杂交,用于GLT - 1 mRNA的35S标记反义寡核苷酸探针在胚胎第11天到成年阶段持续标记套层区/灰质。然而,使用特异性抗体的免疫组织化学在胎儿期和出生后阶段之间的时间内显示出不同的区域和细胞定位。在胎儿期,GLT - 1免疫反应性在边缘区/白质中占主导,在横切面上表现为微小的斑点,在纵切面上表现为细纤维。GLT - 1免疫阳性结构也被神经元特异性烯醇化酶标记,这是一种有丝分裂后神经元和内分泌细胞特有的糖酵解酶。通过电子显微镜观察,在形成频繁膨大的轴突中检测到GLT - 1免疫反应性,并且在一小部分轴膜上呈局灶性定位,特别是面向相邻轴突的部分。在出生后早期,GLT - 1从白质束中的轴突中消失,取而代之的是出现在灰质中围绕各种神经元成分的星形胶质细胞突起中。因此,在转换为星形胶质细胞表达之前,GLT - 1在神经元中短暂表达并定位于分化的轴突中。连同我们之前关于谷氨酸转运体GLAST在放射状胶质纤维中的定位的发现,GLT - 1和GLAST在发育过程中分别定位于不同的定向细胞成分上,年轻神经元的轴突沿着这些成分延伸或其细胞体沿着这些成分移动。