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采用小鼠正中神经模型进行周围神经再生的实验评估。

Employment of the mouse median nerve model for the experimental assessment of peripheral nerve regeneration.

作者信息

Tos P, Ronchi G, Nicolino S, Audisio C, Raimondo S, Fornaro M, Battiston B, Graziani A, Perroteau I, Geuna S

机构信息

UOD di Microchirurgia Ricostruttiva, Dipartimento di Ortopedia e Traumatologia, Ospedale C.T.O., Torino, Italy.

出版信息

J Neurosci Methods. 2008 Mar 30;169(1):119-27. doi: 10.1016/j.jneumeth.2007.11.030. Epub 2007 Dec 8.

DOI:10.1016/j.jneumeth.2007.11.030
PMID:18201767
Abstract

The experimental investigation of nerve regeneration after microsurgical repair is usually carried out in rats, rather than mice, because of the larger sized peripheral nerves. Today however, the availability of genetically modified mice makes the use of this laboratory animal very intriguing for investigating nerve regeneration at a molecular level. In this study we aimed to provide a standardization of the experimental model based on microsurgical direct repair, by 12/0 suture, of the left median nerve in adult male mice. Postoperative recovery was regularly assessed by the grasping test. At day-75 postoperative, regenerated median nerve fibers were analyzed by design-based quantitative morphology and electron microscopy. Yet, sections were immuno-labelled using two axonal antibodies commonly employed for rat nerve fibers. Results indicated that functional recovery begun at day-15 and progressively increased reaching values not significantly different from normal by day-50. Quantitative morphology showed that, at day-75, the number of regenerated nerve fibers was not significantly different in comparison to controls. In contrast, differences were detected in fiber density, mean axon and fiber diameter and myelin thickness which were all significantly lower than controls. Immunohistochemistry showed that axonal markers commonly used for rat nerves studies are effective also for mouse nerves. Similar to the rat, the mouse median nerve model is superior to sciatic nerve model for the minimal impact on animal well-being and the effectiveness of the grasping test for motor function evaluation. The main limitation is the small nerve size which requires advanced microsurgical skills for performing 12/0 epineurial suturing.

摘要

由于周围神经尺寸较大,显微外科修复后神经再生的实验研究通常在大鼠而非小鼠身上进行。然而如今,基因工程小鼠的可得性使得利用这种实验动物在分子水平研究神经再生非常具有吸引力。在本研究中,我们旨在对成年雄性小鼠左侧正中神经采用12/0缝线进行显微外科直接修复的实验模型进行标准化。术后通过抓握试验定期评估恢复情况。术后75天时,通过基于设计的定量形态学和电子显微镜分析再生的正中神经纤维。此外,切片使用通常用于大鼠神经纤维的两种轴突抗体进行免疫标记。结果表明,功能恢复在第15天开始,并逐渐增加,到第50天时达到与正常情况无显著差异的值。定量形态学显示,在第75天时,再生神经纤维的数量与对照组相比无显著差异。相比之下,在纤维密度、平均轴突和纤维直径以及髓鞘厚度方面检测到差异,这些均显著低于对照组。免疫组织化学表明,常用于大鼠神经研究的轴突标记物对小鼠神经也有效。与大鼠类似,小鼠正中神经模型对动物健康的影响最小,且抓握试验对运动功能评估有效,优于坐骨神经模型。主要限制是神经尺寸小,这需要先进的显微外科技术来进行12/0神经外膜缝合。

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