Identification of sensory and motor nerve fascicles by immunofluorescence staining after peripheral nerve injury.
Identification of sensory and motor nerve fascicles by immunofluorescence staining after peripheral nerve injury.
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DOI：
10.1186/s12967-021-02871-w
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发表时间：
2021-05-13
影响因子：
7.4
通讯作者：
Jia X
中科院分区：
文献类型：
--
作者：
Zhou X;Du J;Qing L;Mee T;Xu X;Wang Z;Xu C;Jia X
神经修复或神经移植后运动和感觉纤维的不适当匹配可导致神经恢复失败。运动和感觉纤维的识别对于促进神经再生的新方法的开发和具有感觉反馈技术的下一代神经信号控制的神经假肢的开发是重要的。只有少数几种方法已被报道区分感觉和运动神经束，这些技术的可靠性是未知的。免疫荧光染色是区分感觉和运动神经纤维最常用的方法之一，然而，其准确性仍然未知。在这项研究中，我们的目的是确定流行的免疫荧光标记的运动和感觉神经纤维的疗效。我们收获了大鼠的面神经（主要是运动神经束）和腓肠神经（主要是感觉神经束），并检测了运动标志物（胆碱乙酰转移酶（ChAT），酪氨酸激酶（TrkA））和感觉标志物[神经丝蛋白200 kDa（NF-200），降钙素基因相关肽（CGRP）和瞬时受体电位香草酸亚型1（TRPV 1）]的免疫荧光染色表达。采用平均面积百分比、平均灰度值和轴突计数三种方法对免疫荧光图像中神经标记物的阳性表达进行定量。结果表明，平均灰度值法是最可靠的方法。运动束ChAT和TRKA免疫荧光平均灰度值分别为63.0 ± 0.76%和47.6 ± 0.43%，明显高于感觉束（ChAT：49.2 ± 0.72%，P < 0.001; TRKA：29.1 ± 0.85%，P < 0.001）。此外，TRPV 1的平均灰度值（51.5 ± 0.83%），NF-200运动束的CGRP（61.5 ± 0.62%）和CGRP（37.7 ± 1.22%）明显低于感觉束（71.9 ± 2.32%、69.3 ± 0.46%和54.3 ± 1.04%）（P < 0.001）。使用CHAT/CRCP比率出现最准确的分界点，其中0.855的值具有100%的灵敏度和100%的特异性来识别运动和感觉神经，ROC曲线下面积为1.000（P < 0.001）。ChAT和CGRP的组合建议区分运动和感觉神经纤维。
Inappropriate matching of motor and sensory fibers after nerve repair or nerve grafting can lead to failure of nerve recovery. Identification of motor and sensory fibers is important for the development of new approaches that facilitate neural regeneration and the next generation of nerve signal-controlled neuro-prosthetic limbs with sensory feedback technology. Only a few methods have been reported to differentiate sensory and motor nerve fascicles, and the reliability of these techniques is unknown. Immunofluorescence staining is one of the most commonly used methods to distinguish sensory and motor nerve fibers, however, its accuracy remains unknown. In this study, we aim to determine the efficacy of popular immunofluorescence markers for motor and sensory nerve fibers. We harvested the facial (primarily motor fascicles) and sural (primarily sensory fascicles) nerves in rats, and examined the immunofluorescent staining expressions of motor markers (choline acetyltransferase (ChAT), tyrosine kinase (TrkA)), and sensory markers [neurofilament protein 200 kDa (NF-200), calcitonin gene-related peptide (CGRP) and Transient receptor potential vanillic acid subtype 1 (TRPV1)]. Three methods, including the average area percentage, the mean gray value, and the axon count, were used to quantify the positive expression of nerve markers in the immunofluorescence images. Our results suggest the mean gray value method is the most reliable method. The mean gray value of immunofluorescence in ChAT (63.0 ± 0.76%) and TRKA (47.6 ± 0.43%) on the motor fascicles was significantly higher than that on the sensory fascicles (ChAT: 49.2 ± 0.72%, P < 0.001; and TRKA: 29.1 ± 0.85%, P < 0.001). Additionally, the mean gray values of TRPV1 (51.5 ± 0.83%), NF-200 (61.5 ± 0.62%) and CGRP (37.7 ± 1.22%) on the motor fascicles were significantly lower than that on the sensory fascicles respectively (71.9 ± 2.32%, 69.3 ± 0.46%, and 54.3 ± 1.04%) (P < 0.001). The most accurate cutpoint occurred using CHAT/CRCP ratio, where a value of 0.855 had 100% sensitivity and 100% specificity to identify motor and sensory nerve with an area under the ROC curve of 1.000 (P < 0.001). A combination of ChAT and CGRP is suggested to distinguish motor and sensory nerve fibers.
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影响因子：
3.3
作者：
Castañeda-Corral G;Jimenez-Andrade JM;Bloom AP;Taylor RN;Mantyh WG;Kaczmarska MJ;Ghilardi JR;Mantyh PW
通讯作者：
Mantyh PW
DOI：
10.1002/ar.23843
发表时间：
2018-10-01
影响因子：
2
作者：
Dubovy, Petr;Klusakova, Ilona;Joukal, Marek
通讯作者：
Joukal, Marek
影响因子：
5.3
作者：
Davis-Lopez de Carrizosa, Maria A.;Morado-Diaz, Camilo J.;Pastor, Angel M.
通讯作者：
Pastor, Angel M.
影响因子：
15.9
作者：
Kalin, Stefanie;Miller, Kelly R.;Heppner, Frank L.
通讯作者：
Heppner, Frank L.
影响因子：
3.1
作者：
Choi SI;Lim JY;Yoo S;Kim H;Hwang SW
通讯作者：
Hwang SW