Summary

改良的小鼠神经性疼痛备用神经损伤手术模型

Published: January 25, 2022
doi:

Summary

改良手术是一种用于小鼠或大鼠幸免神经损伤模型的简化方法,只需要一次结扎和一次切割即可损伤腓总神经和腓肠神经。

Abstract

备用神经损伤 (SNI) 是一种模拟周围神经损伤主要症状的动物模型,用于研究小鼠和大鼠神经性疼痛的分子和细胞机制。目前,有两种类型的 SNI 模型,一种是用完整的腓肠神经切割和结扎腓总神经和胫神经,在本研究中定义为 SNIs,另一种是用完整的胫神经切割和结扎腓总神经和腓肠神经,在本研究中定义为 SNIt。因为腓肠神经是纯粹的感觉神经,而胫神经同时包含运动和感觉纤维,所以 SNIt 模型的运动缺陷比 SNIs 模型少得多。在传统的 SNIt 小鼠模型中,腓总神经和腓肠神经分别被切割和结扎。这里描述了一种改进的 SNIt 手术方法,只需一次结扎和一次切割即可损伤腓总神经和腓肠神经,手术时间更短,这更容易执行并降低了拉伸坐骨神经或胫神经的潜在风险,并产生与传统 SNIt 模型相似的机械超敏反应。

Introduction

手术或创伤后神经损伤引起的神经性疼痛具有损害生活质量的重大经济负担。成功开发了许多神经损伤模型,包括脊神经结扎术 (SNL)1、坐骨神经慢性缩窄性损伤 (CCI)2、部分坐骨神经结扎术 (pSNL)3、坐骨神经交易 (SNT)4 和幸存神经损伤 (SNI)5,6,7,8,以模拟大鼠和小鼠周围神经损伤的主要症状,用于研究神经性疼痛 6,7,8,9,10。然而,每种手术模型都有其优点和局限性,因此应特别注意探索和开发手术模型10

啮齿动物 SNI 模型对机械刺激产生长期超敏反应。但是,这有点令人困惑,因为有两种不同的 SNI 模型。最初的 SNI 模型是在 Woolf 的实验室开发的,其中腓总神经和胫神经受损,腓肠神经完好无损 5,6。第二个 SNI 模型是在 Basbaum 的实验室开发的,其中腓总神经和腓肠神经受损,胫神经完好无损 7,8。最初的 Woolf 模型在这里被定义为 SNIs,因为腓肠神经保持完整,而 Basbaum 的模型在这里被定义为 SNIt,因为胫神经保持完整。因为腓肠神经是纯粹的感觉神经,而胫神经同时包含运动和感觉纤维,所以 SNIt 模型的运动缺陷比 SNIs 模型少得多。然而,与 SNIs 模型不同的是,SNIt 模型中的小鼠不会发展为热超敏反应,但在两种模型中都会发展为机械超敏反应。虽然 SNIt 模型是一个相对简单的程序,但它需要分别结扎腓肠神经和腓总神经,并且存在拉伸坐骨神经或胫神经的潜在风险 6,7,8,9。

腓总神经、胫骨神经和腓肠神经是坐骨神经的三个分支,可以在腓肠肌的上缘清楚地识别出来(图 1):胫神经位于腓肠肌下方,腓总神经(头侧)和腓肠神经(尾侧)位于腓肠肌上方11.基于其解剖学特征,开发了一种改良的小鼠 SNIt 手术程序,只需一次神经结扎和一次神经切割即可将腓总神经和腓肠神经结扎在一起,从而缩短了手术时间。

Protocol

动物实验经 UCSF 机构动物护理和使用委员会批准,并根据 NIH 实验动物护理和使用指南进行。本研究使用体重 20-30 g 的成年 C57BL/6 小鼠。von Frey 评估在下午 1:00 到 3:00 之间进行。 1. 麻醉和小鼠准备 将小鼠以 1.0 L/min 的流速放入充满 2% 异氟醚的 O2 溶液中的塑料室中,直至完全麻醉。 使用棉绒涂抹器用眼药膏覆盖?…

Representative Results

改良方法和传统方法之间的手术时间比较。分别在 5 只采用改良方法的小鼠和 5 只采用传统方法的小鼠中记录从切割皮肤开始到闭合皮肤结束的手术时间。使用最少数量的动物来获得具有统计学意义的结果。与传统方法 7,14 执行 SNIt 的对照相比,修改后的方法花费了几乎一半的手术时间 (236.2 ± 28.6 s vs. 4…

Discussion

与分别结扎腓总神经和腓肠神经的传统小鼠 SNIt 方法相比 6,7,8,9,改进的 SNIt 模型具有三个优点:(1) 收缩或拉伸坐骨神经或胫神经的风险较小;(2) 神经切割后无需切除远端神经残端,因为通过将腓总神经和腓肠神经连接在一起,远端神经残端在解剖学上与近端残端分离?…

Divulgazioni

The authors have nothing to disclose.

Acknowledgements

Z.G. 由 NINDS R01NS100801 提供支持。

Materials

6-0 suture Henry Schein 9007482 Nerve ligation and close the muscular layer
Iris Scissors Integra Miltex 12460598 Cut muscle and fascia
Mayo dissecting scissors Fisherbrand 895120 Cut skin incision
Micro forcep Fisherbrand 16100110 Blunt dissection biceps femoris muscle
Micro Scissors Excelta 17467496 Cut nerve
Microdissection Forceps Fisherbrand 16100123 Separate the common peroneal and the sural nerves from the neighboring tissues
Needle Holder Fisherbrand 8966 Hold 6-0 needle
Prism software Graph Pad version 8.0 Statistical analysis software
Wound clips Roboz Surgical NC1878744 Close skin incision

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Citazione di questo articolo
He, L., Zhao, W., Zhang, L., Ilango, M., Zhao, N., Yang, L., Guan, Z. Modified Spared Nerve Injury Surgery Model of Neuropathic Pain in Mice. J. Vis. Exp. (179), e63362, doi:10.3791/63362 (2022).

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