DIFFERENTIAL DISTRIBUTION OF (NA,K)-ATPASE-ALPHA ISOFORM MESSENGER-RNAS IN THE PERIPHERAL NERVOUS-SYSTEM
DIFFERENTIAL DISTRIBUTION OF (NA,K)-ATPASE-ALPHA ISOFORM MESSENGER-RNAS IN THE PERIPHERAL NERVOUS-SYSTEM
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DOI：
10.1016/0006-8993(91)91157-v
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发表时间：
1991-04-12
期刊：
影响因子：
2.9
通讯作者：
FINK, DJ
中科院分区：
文献类型：
--
作者：
MATA, M;SIEGEL, GJ;FINK, DJ
先前已在神经系统中鉴定了（Na，K）-ATP酶α亚基的3种同种型的mRNA转录物（命名为α-1、α-2和α-3）。 为了研究不同α亚型在周围神经系统中的定位和表达，我们从α-1 cDNA的独特3'非翻译区和从α-3 cDNA的翻译区制备探针。 将这些探针用于大鼠脊髓、背根神经节（DRG）和坐骨神经的斑点杂交和原位杂交分析。 在腰脊髓的腹角内，在一组离散的横向放置的运动神经元中发现了α-1 mRNA，而在脊髓的所有识别的神经元中发现了α-3 mRNA，包括含有α-1的运动神经元。 在腰DRG中，α-3均匀分布在DRG神经元中，而α-1在一些神经元中丰富，但在其他神经元中很少或没有发现。 卫星细胞既不包含异构体。 坐骨神经中的雪旺细胞用α-1探针以核周分布标记，但不含可检测的α-3。 斑点杂交显示脊髓和背根神经节中有α 1和α 3，而周围神经中只有α 1。 这些结果表明：（1）核糖探针能够在原位杂交条件下区分α-1和α-3同种型mRNA，（2）在神经元中发现α-1和α-3同种型的（Na，K）-ATP酶，（3）一些运动和感觉神经元含有丰富的α-3，具有很少或不可检测的α-1，而含有丰富的α-1和α-3，（4）在不存在α-3的情况下，腰脊髓或DRG中没有神经元含有可检测的α-1，和（5）雪旺细胞含有α-1同种型，但没有可检测的α-3同种型mRNA。
mRNA transcripts for 3 isoforms of the alpha subunit of (Na,K)-ATPase have been previously identified in the nervous system (designated alpha-1, alpha-2 and alpha-3). In order to study the localization and expression of the different alpha isoforms in the peripheral nervous system, we prepared probes from the unique 3' untranslated region of alpha-1 cDNA, and from the translated region of alpha-3 cDNA. These probes were used in dot blot and in situ hybridization assays of rat spinal cord, dorsal root ganglia (DRG), and sciatic nerve. Within the ventral horn of lumbar spinal cord, alpha-1 mRNA was found in a discrete set of laterally placed motor neurons, while alpha-3 was found in all the identified neurons of the spinal cord, including those motor neurons containing alpha-1. In the lumbar DRG, alpha-3 was uniformly distributed in DRG neurons, while alpha-1 was abundant in some neurons but little or none was found in other neurons. Satellite cells contained neither isoform. Schwann cells in sciatic nerve were labeled with the alpha-1 probe in a perinuclear distribution, but contained no detectable alpha-3. Dot blot analysis showed alpha-1 and alpha-3 in spinal cord and DRG, but only alpha-1 in peripheral nerve. These results imply that: (1) the riboprobes are able to distinguish alpha-1 from alpha-3 isoform mRNAs under in situ hybridization conditions, (2) both alpha-1 and alpha-3 isoforms of (Na,K)-ATPase are found in neurons, (3) some motor and sensory neurons contain abundant alpha-3 with little or no detectable alpha-1, while contain abundant alpha-1 and alpha-3, (4) no neurons in the lumbar spinal cord or DRG contain detectable alpha-1 in the absence of alpha-3, and (5) Schwann cells contain the alpha-1 isoform but no detectable alpha-3 isoform mRNA.