Live imaging of excitable axonal microdomains in ankyrin-G-GFP mice.

IF 3.3 4区管理学 Q2 HOSPITALITY, LEISURE, SPORT & TOURISM Tourist Studies Pub Date : 2024-06-19 DOI:10.1101/2023.02.01.525891

Christian Thome, Jan Maximilian Janssen, Seda Karabulut, Claudio Acuna, Elisa D'Este, Stella J Soyka, Konrad Baum, Michael Bock, Nadja Lehmann, Johannes Roos, Nikolas A Stevens, Masashi Hasegawa, Dan A Ganea, Chloé M Benoit, Jan Gründemann, Lia Min, Kalynn M Bird, Christian Schultz, Vann Bennett, Paul M Jenkins, Maren Engelhardt

{"title":"Live imaging of excitable axonal microdomains in ankyrin-G-GFP mice.","authors":"Christian Thome, Jan Maximilian Janssen, Seda Karabulut, Claudio Acuna, Elisa D'Este, Stella J Soyka, Konrad Baum, Michael Bock, Nadja Lehmann, Johannes Roos, Nikolas A Stevens, Masashi Hasegawa, Dan A Ganea, Chloé M Benoit, Jan Gründemann, Lia Min, Kalynn M Bird, Christian Schultz, Vann Bennett, Paul M Jenkins, Maren Engelhardt","doi":"10.1101/2023.02.01.525891","DOIUrl":null,"url":null,"abstract":"The axon initial segment (AIS) constitutes not only the site of action potential initiation, but also a hub for activity-dependent modulation of output generation. Recent studies shedding light on AIS function used predominantly post-hoc approaches since no robust murine in vivo live reporters exist. Here, we introduce a reporter line in which the AIS is intrinsically labeled by an ankyrin-G-GFP fusion protein activated by Cre recombinase, tagging the native Ank3 gene. Using confocal, superresolution, and two-photon microscopy as well as whole-cell patch-clamp recordings in vitro, ex vivo, and in vivo, we confirm that the subcellular scaffold of the AIS and electrophysiological parameters of labeled cells remain unchanged. We further uncover rapid AIS remodeling following increased network activity in this model system, as well as highly reproducible in vivo labeling of AIS over weeks. This novel reporter line allows longitudinal studies of AIS modulation and plasticity in vivo in real-time and thus provides a unique approach to study subcellular plasticity in a broad range of applications.","PeriodicalId":47199,"journal":{"name":"Tourist Studies","volume":"11 1","pages":""},"PeriodicalIF":3.3000,"publicationDate":"2024-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11212890/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Tourist Studies","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1101/2023.02.01.525891","RegionNum":4,"RegionCategory":"管理学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"HOSPITALITY, LEISURE, SPORT & TOURISM","Score":null,"Total":0}

引用次数: 0

Abstract

The axon initial segment (AIS) constitutes not only the site of action potential initiation, but also a hub for activity-dependent modulation of output generation. Recent studies shedding light on AIS function used predominantly post-hoc approaches since no robust murine in vivo live reporters exist. Here, we introduce a reporter line in which the AIS is intrinsically labeled by an ankyrin-G-GFP fusion protein activated by Cre recombinase, tagging the native Ank3 gene. Using confocal, superresolution, and two-photon microscopy as well as whole-cell patch-clamp recordings in vitro, ex vivo, and in vivo, we confirm that the subcellular scaffold of the AIS and electrophysiological parameters of labeled cells remain unchanged. We further uncover rapid AIS remodeling following increased network activity in this model system, as well as highly reproducible in vivo labeling of AIS over weeks. This novel reporter line allows longitudinal studies of AIS modulation and plasticity in vivo in real-time and thus provides a unique approach to study subcellular plasticity in a broad range of applications.

查看原文

微信好友朋友圈 QQ好友复制链接

本刊更多论文

ankyrin-G-GFP小鼠可兴奋轴突微域的实时成像。

轴突起始节段（AIS）不仅是动作电位的起始部位，也是依赖活动调节输出生成的枢纽。由于没有可靠的小鼠体内活体报告基因，近期揭示 AIS 功能的研究主要采用事后研究的方法。在这里，我们引入了一种报告基因，其中的 AIS 被 Cre 重组酶激活的 ankyrin-G-GFP 融合蛋白内在标记，并标记了原生的 Ank3 基因。利用共焦、超分辨率和双光子显微镜以及体外、体内和体外全细胞膜片钳记录，我们证实 AIS 的亚细胞支架和标记细胞的电生理参数保持不变。我们进一步发现，在这一模型系统中，随着网络活动的增加，AIS 会迅速重塑，而且 AIS 的体内标记可在数周内高度重现。这种新颖的报告基因可以对体内 AIS 的调节和可塑性进行实时纵向研究，从而为研究亚细胞可塑性的广泛应用提供了一种独特的方法。

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分钟内获得全文去求助

来源期刊

Tourist Studies HOSPITALITY, LEISURE, SPORT & TOURISM-

CiteScore

6.90

自引率

4.20%

发文量

期刊介绍： Tourist Studies is a multi-disciplinary journal providing a platform for the development of critical perspectives on the nature of tourism as a social phenomenon through a qualitative lens. Theoretical and multi-disciplinary. Tourist Studies provides a critical social science approach to the study of the tourist and the structures which influence tourist behaviour and the production and reproduction of tourism. The journal examines the relationship between tourism and related fields of social inquiry. Tourism and tourist styles consumption are not only emblematic of many features of contemporary social change, such as mobility, restlessness, the search for authenticity and escape, but they are increasingly central to economic restructuring, globalization, the sociology of consumption and the aestheticization of everyday life. Tourist Studies analyzes these features of tourism from a multi-disciplinary perspective and seeks to evaluate, compare and integrate approaches to tourism from sociology, socio-psychology, leisure studies, cultural studies, geography and anthropology. Global Perspective. Tourist Studies takes a global perspective of tourism, widening and challenging the established views of tourism presented in current periodical literature. Tourist Studies includes: Theoretical analysis with a firm grounding in contemporary problems and issues in tourism studies, qualitative analyses of tourism and the tourist experience, reviews linking theory and policy, interviews with scholars at the forefront of their fields, review essays on particular fields or issues in the study of tourism, review of key texts, publications and visual media relating to tourism studies, and notes on conferences and other events of topical interest to the field of tourism studies.