De León M, Welcher A A, Nahin R H, Liu Y, Ruda M A, Shooter E M, Molina C A
Center for Molecular Biology and Gene Therapy, Loma Linda University, CA 92350, USA.
J Neurosci Res. 1996 May 1;44(3):283-92. doi: 10.1002/(SICI)1097-4547(19960501)44:3<283::AID-JNR9>3.0.CO;2-C.
Peripheral nerve trauma induces the expression of genes presumed to be involved in the process of nerve degeneration and repair. In the present study, an in vivo paradigm was employed to identify molecules which may have important roles in these processes. A cDNA library was constructed with RNA extracted from rat dorsal root ganglia (DRG) 3 days after a sciatic nerve crush. After differential hybridization to this library, several cDNAs were identified that encoded mRNAs that were upregulated in the DRG ipsilateral to the crush injury, as opposed to the contralateral or naive DRG. Approximately 0.15% of all the clones screened were found to be induced. This report presents the types of induced sequences identified and characterizes one of them, DA11. The 0.7 kb DA11 full length cDNA clone contains a 405 nucleotide open reading frame that encodes a putative protein of 15.2 kDa (135 amino acid residues) and is a member of the family of fatty acid binding proteins (FABP). The DA11 protein differs by one amino acid residue from the sequence of the C-FAPB protein and by eight residues from the sequence of mal1, proteins found in rat and mouse skin, respectively. Northern and Western blot analyses showed that the DA11 mRNA and protein were induced in the injured DRG. Furthermore, studies using antibodies generated against DA11 found that the DA11-like immunoreactivity was more pronounced in the nuclei of neurons located in the DRG ipsilateral to the sciatic cut than those located in the contralateral DRG. The induction of DA11 mRNA and protein in DRG neurons suggests, for the first time, the involvement of a neuronal FABP in the process of degeneration and repair in the nervous system.
外周神经损伤会诱导一些被认为参与神经退变和修复过程的基因表达。在本研究中,采用了一种体内实验模式来鉴定可能在这些过程中起重要作用的分子。用坐骨神经挤压3天后大鼠背根神经节(DRG)提取的RNA构建了一个cDNA文库。与该文库进行差异杂交后,鉴定出了几个cDNA,它们编码的mRNA在挤压损伤同侧的DRG中上调,而在对侧或未损伤的DRG中未上调。在所有筛选的克隆中,约0.15%被发现是诱导型的。本报告介绍了所鉴定的诱导序列类型,并对其中一个名为DA11的序列进行了表征。0.7 kb的DA11全长cDNA克隆包含一个405个核苷酸的开放阅读框,编码一个推定的15.2 kDa蛋白(135个氨基酸残基),是脂肪酸结合蛋白(FABP)家族的成员。DA11蛋白与C-FAPB蛋白序列有一个氨基酸残基的差异,并与大鼠和小鼠皮肤中分别发现的mal1蛋白序列有八个残基的差异。Northern和Western印迹分析表明,DA11 mRNA和蛋白在损伤的DRG中被诱导。此外,使用针对DA11产生的抗体进行的研究发现,与对侧DRG中的神经元相比,坐骨神经切断同侧DRG中神经元细胞核中的DA11样免疫反应性更明显。DRG神经元中DA11 mRNA和蛋白的诱导首次表明神经元FABP参与了神经系统的退变和修复过程。