Wensley C H, Stone D M, Baker H, Kauer J S, Margolis F L, Chikaraishi D M
Molecular Biology and Microbiology Department, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
J Neurosci. 1995 Jul;15(7 Pt 1):4827-37. doi: 10.1523/JNEUROSCI.15-07-04827.1995.
The separation between the cell bodies of olfactory receptor neurons in the nasal cavity and their axon terminals in the olfactory bulb make them attractive for studying axonal transport. Although high molecular weight RNAs are generally believed to be excluded from axons of mature neurons, we demonstrate here that mRNA for olfactory marker protein (OMP), an abundant cytoplasmic protein selectively expressed in mature receptor cells, is present in rodent olfactory receptor axons. OMP RNA was detected by in situ hybridization at the light microscope level in axons and in terminals. By nuclease protection, the level of OMP RNA in the olfactory bulb was 5-10% of that in the olfactory epithelium where the cell bodies reside. In contrast to axonally transported vasopressin and oxytocin mRNAs, which are deficient in their 3' polyA tails, axonal OMP RNA fractionated as polyA+. OMP RNA was lost from axons and terminals after deafferentation, suggesting that OMP RNA was synthesized in receptor cell bodies in the epithelium and was transported into axons and terminals in the olfactory bulb. RNA for G(olf), a G-protein highly expressed in dendrites of mature olfactory receptor neurons, was not detected in the olfactory bulb. We hypothesize that the immature nature of the cytoskeleton and, specifically, the lack of tightly bundled microtubules allows transport of particular mRNAs in olfactory receptor axons.
鼻腔中嗅觉受体神经元的细胞体与其在嗅球中的轴突终末之间的分隔,使得它们成为研究轴突运输的理想对象。尽管一般认为高分子量RNA会被排除在成熟神经元的轴突之外,但我们在此证明,嗅觉标记蛋白(OMP)的mRNA存在于啮齿动物的嗅觉受体轴突中,OMP是一种在成熟受体细胞中选择性表达的丰富细胞质蛋白。通过光学显微镜水平的原位杂交在轴突和终末中检测到了OMP RNA。通过核酸酶保护实验发现,嗅球中OMP RNA的水平是其细胞体所在的嗅上皮中OMP RNA水平的5%-10%。与轴突运输的血管加压素和催产素mRNA不同,后者在其3'多聚腺苷尾存在缺陷,轴突中的OMP RNA被分离为多聚腺苷酸阳性。去传入神经后,轴突和终末中的OMP RNA消失,这表明OMP RNA是在上皮中的受体细胞体中合成的,并被运输到嗅球的轴突和终末中。在嗅球中未检测到G(olf)的RNA,G(olf)是一种在成熟嗅觉受体神经元树突中高度表达的G蛋白。我们推测,细胞骨架的不成熟性质,特别是缺乏紧密排列的微管,使得特定的mRNA能够在嗅觉受体轴突中运输。