ISOLATION OF CDNA CLONES ENCODING RAT GLIAL FIBRILLARY ACIDIC PROTEIN - EXPRESSION IN ASTROCYTES AND IN SCHWANN-CELLS
ISOLATION OF CDNA CLONES ENCODING RAT GLIAL FIBRILLARY ACIDIC PROTEIN - EXPRESSION IN ASTROCYTES AND IN SCHWANN-CELLS
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DOI：
10.1002/jnr.490320102
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发表时间：
1992-05-01
影响因子：
4.2
通讯作者：
MILNER, RJ
中科院分区：
文献类型：
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作者：
FEINSTEIN, DL;WEINMASTER, GA;MILNER, RJ
中枢神经系统(CNS)中星形胶质细胞表达的胶质纤维酸性蛋白(GFAP)已被广泛研究，但周围神经系统(PNS)中相关分子的分子同一性尚不清楚。为了研究CNS和PNS GFAP之间可能的结构差异，我们从培养的星形胶质细胞和雪旺细胞库中分离了大鼠GFAP的cDNA克隆。核苷酸序列分析表明，PNS和CNS GFAP克隆含有相同的编码区，预测的蛋白质产物为430个氨基酸。然而，从雪旺细胞文库中分离的克隆rGFA15的5‘非翻译区比预测的脑源性GFAP mRNA要长。从RT4-D6雪旺细胞系中分离的RNA的引物延伸分析表明，PNS GFAP mRNA的起始点位于中枢神经系统的上游169个碱基。此外，从培养的星形胶质细胞和雪旺细胞制备的GFAP的胰酶肽图谱显示，CNS GFAP中有一个主要的肽片段，而PNS GFAP中没有。这些结果表明这两种细胞的GFAP在核酸和蛋白质水平上都存在结构差异，这与先前观察到的CNS和PNS GFAP之间存在的免疫化学差异是一致的。
Glial fibrillary acidic protein (GFAP) expressed by astrocytes in the central nervous system (CNS) has been extensively characterized but the molecular identity of related molecules in the peripheral nervous system (PNS) remains unclear. To examine possible structural differences between CNS and PNS GFAP, we have isolated cDNA clones for rat GFAP from both cultured astrocyte and Schwann cell libraries. Nucleotide sequence analysis indicated that the PNS and CNS GFAP clones contained identical coding regions, with a predicted protein product of 430 amino acids. However, the 5'-untranslated region of clone rGFA15, isolated from the Schwann cell library, was longer than that predicted for brain-derived GFAP mRNA. Primer extension analysis of RNA isolated from the RT4-D6 Schwann cell line indicated that the start site for PNS GFAP mRNA lies 169 bases upstream from that used in the CNS. In addition, tryptic peptide mapping of GFAP prepared from cultured astrocytes and Schwann cells revealed one major peptide fragment present in CNS GFAP but absent from PNS GFAP. These results suggest structural differences between GFAP in these two cell types, at both the nucleic acid and protein level, and are consistent with previous observations of immunochemical differences existing between CNS and PNS GFAP.