Moscoso L M, Sanes J R
Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
J Comp Neurol. 1995 Feb 13;352(3):321-34. doi: 10.1002/cne.903520302.
To identify cell adhesion molecules (CAMs) expressed by mammalian motoneurons, we applied the polymerase chain reaction to a murine motor neuron-like cell line, NSC-34. Using primers derived from a group of L1-related CAMs, we cloned two alternatively spliced forms of mouse L1, which differ by a 12-base-pair insert, plus putative murine orthologs of the chicken cell adhesion molecules Nr-CAM/Bravo and neurofascin. All four mRNAs are expressed in NSC-34 cells, but only neurofascin and the insert-minus form of L1 are expressed in its neuroblastoma parent, N18TG2. Analysis of RNA in neonatal tissues reveals expression largely restricted to the brain and spinal cord. In situ hybridization histochemistry of spinal cord shows that motoneurons express L1, Nr-CAM, and neurofascin as well as N-CAM. L1 and N-CAM RNAs are detected throughout the period studied (from embryonic day [E]11 to postnatal day [P]28), whereas Nr-CAM is expressed only at early ages (< E15) and neurofascin is predominantly expressed postnatally. Moreover, each CAM is expressed by distinct subsets of neighboring cells and at distinct times. For example, Nr-CAM mRNA is present in floor plate cells of embryonic spinal cord, whereas neurofascin is expressed by a subset of glia postnatally. Finally, we show that each CAM has a distinct spatiotemporal pattern of expression in dorsal root ganglia.
为了鉴定哺乳动物运动神经元所表达的细胞黏附分子(CAMs),我们将聚合酶链反应应用于一种小鼠运动神经元样细胞系NSC-34。使用源自一组与L1相关的CAMs的引物,我们克隆了小鼠L1的两种可变剪接形式,它们相差12个碱基对的插入片段,此外还克隆了鸡细胞黏附分子Nr-CAM/Bravo和神经束蛋白的假定小鼠直系同源物。所有这四种mRNA在NSC-34细胞中均有表达,但在其神经母细胞瘤亲本N18TG2中仅表达神经束蛋白和L1的无插入片段形式。对新生组织中的RNA分析显示,表达主要局限于脑和脊髓。脊髓的原位杂交组织化学表明,运动神经元表达L1、Nr-CAM、神经束蛋白以及N-CAM。在整个研究期间(从胚胎第[E]11天到出生后第[P]28天)均检测到L1和N-CAM的RNA,而Nr-CAM仅在早期(<E15)表达,神经束蛋白主要在出生后表达。此外,每种CAM由相邻细胞的不同亚群在不同时间表达。例如,Nr-CAM mRNA存在于胚胎脊髓的底板细胞中,而神经束蛋白在出生后由一部分神经胶质细胞表达。最后,我们表明每种CAM在背根神经节中具有独特的时空表达模式。