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鱿鱼巨轴突合成 NF 蛋白。

Squid Giant Axons Synthesize NF Proteins.

机构信息

Department of Biology, University of Naples Federico II, Via Cinthia, 80126, Naples, Italy.

Department of Biology and Evolution of Marine Organisms, Stazione Zoologica Anton Dohrn, Villa Comunale 1, Napoli, 80121, Italy.

出版信息

Mol Neurobiol. 2018 Apr;55(4):3079-3084. doi: 10.1007/s12035-017-0561-z. Epub 2017 May 2.

Abstract

Squid giant axon has been an excellent model system for studying fundamental topics in neurobiology such as neuronal signaling. It has been also useful in addressing the questions of local protein synthesis in the axons. Incubation of isolated squid giant axons with [S]methionine followed by immunoprecipitation with a rabbit antibody against all squid neurofilament (NF) proteins demonstrates the local synthesis of a major 180 kDa NF protein and of several NF proteins of lower molecular weights. Their identification as NF proteins is based on their absence in the preimmune precipitates. Immunoprecipitates washed with more stringent buffers confirmed these results. Our data are at variance with a recent study based on the same experimental procedure that failed to visualize the local synthesis of NF proteins by the giant axon and thereby suggested their exclusive derivation from nerve cell bodies (as reported by Gainer et al. in Cell Mol Neurobiol 37:475-486, 2017). By reviewing the pertinent literature, we confute the claims that mRNA translation is absent in mature axons because of a putative translation block and that most proteins of mature axons are synthesized in the surrounding glial cells. Given the intrinsic axonal capacity to synthesize proteins, we stress the glial derivation of axonal and presynaptic RNAs and the related proposal that these neuronal domains are endowed with largely independent gene expression systems (as reported by Giuditta et al. in Physiol Rev 88:515-555, 2008).

摘要

鱿鱼巨大轴突一直是研究神经生物学基本课题(如神经元信号传递)的极佳模式系统。它也有助于解决轴突中局部蛋白质合成的问题。用[S]甲硫氨酸孵育分离的鱿鱼巨大轴突,然后用针对所有鱿鱼神经丝(NF)蛋白的兔抗体进行免疫沉淀,证明了一种主要的 180 kDa NF 蛋白和几种分子量较低的 NF 蛋白的局部合成。它们被鉴定为 NF 蛋白是基于它们在未免疫沉淀物中的不存在。用更严格的缓冲液洗涤免疫沉淀物证实了这些结果。我们的数据与最近的一项基于相同实验程序的研究结果不一致,该研究未能观察到巨大轴突中 NF 蛋白的局部合成,因此暗示它们仅源自神经细胞体(正如 Gainer 等人在 Cell Mol Neurobiol 37:475-486,2017 年报道的那样)。通过回顾相关文献,我们驳斥了成熟轴突中由于所谓的翻译阻断而不存在 mRNA 翻译以及成熟轴突中的大多数蛋白质都是在周围神经胶质细胞中合成的观点。鉴于轴突具有内在的蛋白质合成能力,我们强调了胶质细胞衍生的轴突和突触前 RNA,以及相关的提议,即这些神经元区域具有很大程度上独立的基因表达系统(正如 Giuditta 等人在 Physiol Rev 88:515-555,2008 年报道的那样)。

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