Paradies M A, Steward O
Department of Neuroscience, University of Virginia School of Medicine, Charlottesville 22908, USA.
J Neurobiol. 1997 Oct;33(4):473-93. doi: 10.1002/(sici)1097-4695(199710)33:4<473::aid-neu10>3.0.co;2-d.
Previous studies have established that most of the mRNAs that neurons express are localized in the cell body and very proximal dendrites, whereas a small subset of mRNAs is present at relatively high levels in dendrites. It is not clear, however, whether particular mRNAs have the same subcellular distribution in different types of neurons or whether different types of neurons sort mRNAs in different ways. The present study was undertaken to address these questions. Nonisotopic in situ hybridization techniques were used to define the subcellular localization of representative mRNAs including beta-tubulin, low-molecular-weight neurofilament protein (NF-68), high-molecular-weight microtubule-associated protein (MAP2), growth-associated protein 43 (F1/GAP43), the alpha subunit of calcium/calmodulin-dependent protein kinase II (alpha CaMII kinase), and poly (A+) mRNA. The mRNAs for beta-tubulin, neurofilament 68, and F1/GAP43 were restricted to the region of the cell body and very proximal dendrites in most neurons. In some neuron types, however, labeling for NF-68 extended for considerable distances into dendrites. In some neurons that express MAP2, the mRNA was present at the highest levels in the proximal third to half of the dendritic arbor, whereas in other neurons the highest levels of labeling were in the cell body. In most neurons that express alpha CaMII kinase, the highest levels of the mRNA were in the cell body, but labeling was also present throughout dendrites. However, in a few types of neurons, alpha CaMII kinase mRNA was largely restricted to the cell body. The fact that there are no general rules for mRNA localization that apply to all neuron types implies the existence of neuron type-specific mechanisms that regulate mRNA distribution.
以往的研究表明,神经元表达的大多数mRNA定位于细胞体和非常靠近细胞体的树突中,而一小部分mRNA在树突中的含量相对较高。然而,尚不清楚特定的mRNA在不同类型的神经元中是否具有相同的亚细胞分布,或者不同类型的神经元是否以不同的方式对mRNA进行分类。本研究旨在解决这些问题。采用非同位素原位杂交技术来确定代表性mRNA的亚细胞定位,这些mRNA包括β-微管蛋白、低分子量神经丝蛋白(NF-68)、高分子量微管相关蛋白(MAP2)、生长相关蛋白43(F1/GAP43)、钙/钙调蛋白依赖性蛋白激酶II的α亚基(α CaMII激酶)和聚腺苷酸(A+)mRNA。β-微管蛋白、神经丝68和F1/GAP43的mRNA在大多数神经元中局限于细胞体区域和非常靠近细胞体的树突中。然而,在某些神经元类型中,NF-68的标记延伸到树突中的相当远距离。在一些表达MAP2的神经元中,mRNA在树突分支近端三分之一到一半的区域含量最高,而在其他神经元中,标记的最高水平在细胞体中。在大多数表达α CaMII激酶的神经元中,mRNA的最高水平在细胞体中,但树突中也有标记。然而,在少数几种神经元中,α CaMII激酶mRNA主要局限于细胞体。不存在适用于所有神经元类型的mRNA定位通用规则这一事实意味着存在调节mRNA分布的神经元类型特异性机制。