Spinal cord trunk preparation for analyzing cross-segmental primary afferent signal transmission and modulation

IF 2.7 4区 医学 Q2 BIOCHEMICAL RESEARCH METHODS
Caifeng Shao , Mingwei Zhao , Kun Yang
{"title":"Spinal cord trunk preparation for analyzing cross-segmental primary afferent signal transmission and modulation","authors":"Caifeng Shao ,&nbsp;Mingwei Zhao ,&nbsp;Kun Yang","doi":"10.1016/j.jneumeth.2025.110440","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><div>The spinal cord dorsal horn is pivotal for primary afferent signal transmission and modulation. Primary afferent fibers from each dorsal root arrive at the dorsal horn and travel 1–2 segments caudally and rostrally. Usually, in vitro spinal cord slices or in vivo preparations are employed for primary afferent stimulation and patch-clamp recordings to assess input signals. However, the spinal cord slices lose \"intact\" cross-segmental pathways, and in vivo studies are technically challenging.</div></div><div><h3>New method</h3><div>Here, we describe the preparation of a spinal cord trunk for analyzing afferent signal cross-segmental transmission in adult rats. By combining patch-clamp recording, Lissauer's tract stimulation, and ambient temperature manipulation, our methods enable accessing primary afferent pathways within several segments.</div></div><div><h3>Results</h3><div>Our present spinal trunk preparation can be maintained healthy for about 5 h. Lissauer’s tract stimulation induced evoked excitatory postsynaptic currents (eEPSCs) recorded in 6–10 mm rostrally in the ipsilateral dorsal horn. The eEPSCs, spontaneous EPSCs (sEPSCs), and neural excitability can be modulated by ambient temperature rise. Neuropharmacological studies can also be conducted on this spinal trunk preparation.</div></div><div><h3>Compared with existing methods</h3><div>Compared with conventional in vitro spinal cord slices, our present method maintains a relatively intact cross-segment pathway in the dorsal horn; compared with in vivo study, it avoids mechanical vibration and other technical challenges in living animals.</div></div><div><h3>Conclusion</h3><div>The rodent spinal cord trunk can be maintained for an extended period in a fully submerged chamber; combined with patch clamp recordings, our protocol facilitates the study of primary afferent transmission and modulation in the dorsal horn within adjacent segments.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"418 ","pages":"Article 110440"},"PeriodicalIF":2.7000,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Neuroscience Methods","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0165027025000810","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0

Abstract

Background

The spinal cord dorsal horn is pivotal for primary afferent signal transmission and modulation. Primary afferent fibers from each dorsal root arrive at the dorsal horn and travel 1–2 segments caudally and rostrally. Usually, in vitro spinal cord slices or in vivo preparations are employed for primary afferent stimulation and patch-clamp recordings to assess input signals. However, the spinal cord slices lose "intact" cross-segmental pathways, and in vivo studies are technically challenging.

New method

Here, we describe the preparation of a spinal cord trunk for analyzing afferent signal cross-segmental transmission in adult rats. By combining patch-clamp recording, Lissauer's tract stimulation, and ambient temperature manipulation, our methods enable accessing primary afferent pathways within several segments.

Results

Our present spinal trunk preparation can be maintained healthy for about 5 h. Lissauer’s tract stimulation induced evoked excitatory postsynaptic currents (eEPSCs) recorded in 6–10 mm rostrally in the ipsilateral dorsal horn. The eEPSCs, spontaneous EPSCs (sEPSCs), and neural excitability can be modulated by ambient temperature rise. Neuropharmacological studies can also be conducted on this spinal trunk preparation.

Compared with existing methods

Compared with conventional in vitro spinal cord slices, our present method maintains a relatively intact cross-segment pathway in the dorsal horn; compared with in vivo study, it avoids mechanical vibration and other technical challenges in living animals.

Conclusion

The rodent spinal cord trunk can be maintained for an extended period in a fully submerged chamber; combined with patch clamp recordings, our protocol facilitates the study of primary afferent transmission and modulation in the dorsal horn within adjacent segments.
脊髓干准备分析跨节段初级传入信号的传输和调制
脊髓背角对初级传入信号的传递和调制起关键作用。各背根的初级传入纤维到达背角,沿尾侧和喙侧行进1-2节。通常,体外脊髓切片或体内制剂用于初级传入刺激和膜片钳记录以评估输入信号。然而,脊髓切片失去了“完整的”跨节段通路,体内研究在技术上具有挑战性。新方法在此,我们描述了用于分析成年大鼠传入信号跨节段传递的脊髓干的制备。通过结合膜片钳记录、利索耳束刺激和环境温度操纵,我们的方法能够访问几个片段内的主要传入通路。结果本实验制备的脊髓干可保持健康5 h左右。利索道刺激诱发的兴奋性突触后电流(eEPSCs)记录在同侧背角背侧6-10 mm。EPSCs、自发EPSCs (sEPSCs)和神经兴奋性可以通过环境温度升高来调节。神经药理学研究也可以对这种脊髓干制剂进行。与传统的体外脊髓切片相比,我们的方法在脊髓背角中保持了相对完整的横段通路;与体内研究相比,它避免了活体动物的机械振动和其他技术挑战。结论鼠类脊髓干在全浸没腔中可长时间保持;结合膜片钳记录,我们的协议有助于研究相邻段内背角的主要传入传输和调制。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Journal of Neuroscience Methods
Journal of Neuroscience Methods 医学-神经科学
CiteScore
7.10
自引率
3.30%
发文量
226
审稿时长
52 days
期刊介绍: The Journal of Neuroscience Methods publishes papers that describe new methods that are specifically for neuroscience research conducted in invertebrates, vertebrates or in man. Major methodological improvements or important refinements of established neuroscience methods are also considered for publication. The Journal''s Scope includes all aspects of contemporary neuroscience research, including anatomical, behavioural, biochemical, cellular, computational, molecular, invasive and non-invasive imaging, optogenetic, and physiological research investigations.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信