*Result*: Live imaging of excitable axonal microdomains in ankyrin-G-GFP mice.
Title:
Live imaging of excitable axonal microdomains in ankyrin-G-GFP mice.
Authors:
Thome C; Institute of Anatomy and Cell Biology, Johannes Kepler University, Linz, Austria.; Clinical Research Institute for Neurosciences, Johannes Kepler University, Linz, Austria.; Institute of Physiology and Pathophysiology, Heidelberg University, Heidelberg, Germany., Janssen JM; Institute of Anatomy and Cell Biology, Johannes Kepler University, Linz, Austria.; Clinical Research Institute for Neurosciences, Johannes Kepler University, Linz, Austria.; Institute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany., Karabulut S; Institute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany., Acuna C; Chica and Heinz Schaller Research Group, Institute of Anatomy and Cell Biology, Heidelberg University, Heidelberg, Germany., D'Este E; Optical Microscopy Facility, Max Planck Institute for Medical Research, Heidelberg, Germany., Soyka SJ; Institute of Anatomy and Cell Biology, Dept. of Functional Neuroanatomy, Heidelberg University, Heidelberg, Germany., Baum K; Institute of Anatomy and Cell Biology, Johannes Kepler University, Linz, Austria.; Clinical Research Institute for Neurosciences, Johannes Kepler University, Linz, Austria., Bock M; Institute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany., Lehmann N; Institute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany., Roos J; Institute of Anatomy and Cell Biology, Johannes Kepler University, Linz, Austria.; Clinical Research Institute for Neurosciences, Johannes Kepler University, Linz, Austria.; Institute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany., Stevens NA; Institute of Physiology and Pathophysiology, Heidelberg University, Heidelberg, Germany., Hasegawa M; German Center for Neurodegenerative Disease (DZNE), Neural Circuit Computations, Bonn, Germany., Ganea DA; Department of Biomedicine, University of Basel, Basel, Switzerland., Benoit CM; German Center for Neurodegenerative Disease (DZNE), Neural Circuit Computations, Bonn, Germany.; Department of Biomedicine, University of Basel, Basel, Switzerland., Gründemann J; German Center for Neurodegenerative Disease (DZNE), Neural Circuit Computations, Bonn, Germany.; Department of Biomedicine, University of Basel, Basel, Switzerland., Min LY; Department of Pharmacology, University of Michigan Medical School, Ann Arbor, United States., Bird KM; Department of Pharmacology, University of Michigan Medical School, Ann Arbor, United States., Schultz C; Institute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany., Bennett V; Department of Biochemistry, Duke University Medical Center, Durham, United States., Jenkins PM; Department of Pharmacology, University of Michigan Medical School, Ann Arbor, United States.; Department of Psychiatry, University of Michigan Medical School, Ann Arbor, United States., Engelhardt M; Institute of Anatomy and Cell Biology, Johannes Kepler University, Linz, Austria.; Clinical Research Institute for Neurosciences, Johannes Kepler University, Linz, Austria.; Institute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
Source:
ELife [Elife] 2025 Feb 03; Vol. 12. Date of Electronic Publication: 2025 Feb 03.
Publication Type:
Journal Article
Language:
English
Journal Info:
Publisher: eLife Sciences Publications, Ltd Country of Publication: England NLM ID: 101579614 Publication Model: Electronic Cited Medium: Internet ISSN: 2050-084X (Electronic) Linking ISSN: 2050084X NLM ISO Abbreviation: Elife Subsets: MEDLINE
Imprint Name(s):
Original Publication: Cambridge, UK : eLife Sciences Publications, Ltd., 2012-
MeSH Terms:
Ankyrins*/genetics , Ankyrins*/metabolism , Axons*/physiology , Axon Initial Segment*/physiology, Green Fluorescent Proteins/genetics ; Green Fluorescent Proteins/metabolism ; Recombinant Fusion Proteins/genetics ; Recombinant Fusion Proteins/metabolism ; Animals ; Mice ; Mice, Transgenic ; Genes, Reporter ; Patch-Clamp Techniques ; Microscopy, Confocal ; Action Potentials
Comments:
Update of: bioRxiv. 2024 Jun 19:2023.02.01.525891. doi: 10.1101/2023.02.01.525891.. (PMID: 38948770)
Erratum in: Elife. 2025 Sep 11;14:e109210. doi: 10.7554/eLife.109210.. (PMID: 40932473)
Erratum in: Elife. 2025 Sep 11;14:e109210. doi: 10.7554/eLife.109210.. (PMID: 40932473)
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Nature. 2010 Jun 24;465(7301):1075-8. (PMID: 20543825)
J Cell Biol. 2007 Aug 27;178(5):875-86. (PMID: 17709431)
Proc Natl Acad Sci U S A. 2017 Jan 10;114(2):E191-E199. (PMID: 28003466)
Nat Biotechnol. 2003 May;21(5):562-5. (PMID: 12665802)
Nat Rev Neurosci. 2021 Jan;22(1):7-20. (PMID: 33239761)
Cell Rep. 2015 Dec 29;13(12):2781-93. (PMID: 26711344)
J Neurosci. 2016 Apr 20;36(16):4421-33. (PMID: 27098687)
Proc Natl Acad Sci U S A. 2015 Jan 27;112(4):1214-9. (PMID: 25552561)
Front Cell Neurosci. 2019 Jan 22;13:1. (PMID: 30723396)
Neuron. 2007 Aug 16;55(4):633-47. (PMID: 17698015)
J Comp Neurol. 2007 Jan 10;500(2):339-52. (PMID: 17111377)
Annu Rev Neurosci. 2012;35:249-65. (PMID: 22443507)
Neuron. 2019 Oct 23;104(2):305-321.e8. (PMID: 31474508)
Proc Natl Acad Sci U S A. 2002 Feb 19;99(4):2303-8. (PMID: 11842202)
J Neurosci. 2008 Dec 31;28(53):14329-40. (PMID: 19118165)
Front Cell Neurosci. 2019 Oct 15;13:456. (PMID: 31680869)
J Neurosci. 2000 Nov 15;20(22):8354-64. (PMID: 11069942)
J Cell Biol. 2004 Sep 27;166(7):983-90. (PMID: 15381686)
Trends Neurosci. 2016 Oct;39(10):656-667. (PMID: 27637565)
Proc Natl Acad Sci U S A. 2017 Jan 3;114(1):154-159. (PMID: 27994149)
J Cell Biol. 1998 Sep 21;142(6):1571-81. (PMID: 9744885)
Sci Adv. 2018 Nov 28;4(11):eaau8621. (PMID: 30498783)
Nature. 2010 Jun 24;465(7301):1070-4. (PMID: 20543823)
Neuron. 2012 Jan 26;73(2):235-47. (PMID: 22284179)
Appl Opt. 1983 Feb 1;22(3):396-9. (PMID: 18195798)
Front Cell Neurosci. 2017 Jun 06;11:157. (PMID: 28634442)
Nat Rev Neurosci. 2010 Aug;11(8):552-62. (PMID: 20631711)
Sci Adv. 2022 Sep 9;8(36):eabo6333. (PMID: 36083912)
Nat Neurosci. 2009 Aug;12(8):996-1002. (PMID: 19633666)
Proc Natl Acad Sci U S A. 2015 Aug 4;112(31):9757-62. (PMID: 26195803)
J Neurosci. 2019 Jun 19;39(25):4864-4873. (PMID: 30967428)
J Biol Chem. 2012 Dec 21;287(52):43995-4005. (PMID: 23129772)
Sci Adv. 2023 Sep 15;9(37):eadf3885. (PMID: 37713493)
Proc Natl Acad Sci U S A. 2021 Nov 23;118(47):. (PMID: 34799447)
Front Cell Neurosci. 2019 Jul 30;13:318. (PMID: 31417359)
Brain Struct Funct. 2018 Nov;223(8):3855-3873. (PMID: 30094604)
Cereb Cortex. 2017 Sep 1;27(9):4662-4675. (PMID: 28922860)
Elife. 2020 Jun 17;9:. (PMID: 32553116)
J Neurosci. 2020 Oct 28;40(44):8413-8425. (PMID: 32978288)
Nat Commun. 2021 Jan 4;12(1):23. (PMID: 33397944)
J Cell Biol. 2007 Jun 4;177(5):857-70. (PMID: 17548513)
J Physiol. 2018 Aug;596(16):3759-3773. (PMID: 29873393)
Front Cell Neurosci. 2016 Nov 23;10:268. (PMID: 27932952)
Nat Commun. 2019 Sep 19;10(1):4264. (PMID: 31537802)
Neuron. 2014 Oct 22;84(2):399-415. (PMID: 25374361)
J Biol Chem. 2013 May 17;288(20):14018-14031. (PMID: 23530049)
Elife. 2017 Jan 28;6:. (PMID: 28130923)
J Neurosci. 2015 Feb 4;35(5):2283-92. (PMID: 25653382)
J Cell Biol. 2000 Nov 27;151(5):985-1002. (PMID: 11086001)
Exp Neurol. 2018 Apr;302:169-180. (PMID: 29305051)
Brain Struct Funct. 2014 Jul;219(4):1433-50. (PMID: 23728480)
J Biol Chem. 2004 Apr 16;279(16):16706-14. (PMID: 14757759)
Hippocampus. 2010 Apr;20(4):558-71. (PMID: 19603521)
J Comp Neurol. 2007 Oct 10;504(5):441-9. (PMID: 17701995)
J Cell Biol. 2002 Jan 21;156(2):337-48. (PMID: 11807096)
Cereb Cortex. 2012 Jul;22(7):1648-61. (PMID: 21940706)
Nat Neurosci. 2008 Feb;11(2):178-86. (PMID: 18204443)
Neuroscience. 2018 Jan 1;368:268-282. (PMID: 28739523)
Proc Natl Acad Sci U S A. 2006 Feb 21;103(8):2920-5. (PMID: 16473933)
Front Cell Neurosci. 2017 Nov 06;11:332. (PMID: 29170630)
J Cell Biol. 2014 Jul 21;206(2):273-88. (PMID: 25049274)
Proc Natl Acad Sci U S A. 2015 Jan 27;112(4):957-64. (PMID: 25552556)
J Cell Biol. 1998 Nov 30;143(5):1295-304. (PMID: 9832557)
Front Neuroanat. 2018 Sep 11;12:74. (PMID: 30254572)
Mol Psychiatry. 2020 Nov;25(11):2800-2817. (PMID: 30504823)
Front Neuroanat. 2014 Mar 11;8:11. (PMID: 24653680)
Front Cell Neurosci. 2016 Oct 25;10:250. (PMID: 27826229)
J Neurosci. 2018 Feb 28;38(9):2135-2145. (PMID: 29378864)
Neuron. 2009 Jan 29;61(2):259-71. (PMID: 19186168)
J Neurosci. 2020 Aug 26;40(35):6709-6721. (PMID: 32719016)
Front Cell Neurosci. 2017 May 09;11:136. (PMID: 28536506)
Nature. 2010 Jun 24;465(7301):1075-8. (PMID: 20543825)
J Cell Biol. 2007 Aug 27;178(5):875-86. (PMID: 17709431)
Proc Natl Acad Sci U S A. 2017 Jan 10;114(2):E191-E199. (PMID: 28003466)
Nat Biotechnol. 2003 May;21(5):562-5. (PMID: 12665802)
Nat Rev Neurosci. 2021 Jan;22(1):7-20. (PMID: 33239761)
Cell Rep. 2015 Dec 29;13(12):2781-93. (PMID: 26711344)
J Neurosci. 2016 Apr 20;36(16):4421-33. (PMID: 27098687)
Proc Natl Acad Sci U S A. 2015 Jan 27;112(4):1214-9. (PMID: 25552561)
Front Cell Neurosci. 2019 Jan 22;13:1. (PMID: 30723396)
Neuron. 2007 Aug 16;55(4):633-47. (PMID: 17698015)
J Comp Neurol. 2007 Jan 10;500(2):339-52. (PMID: 17111377)
Annu Rev Neurosci. 2012;35:249-65. (PMID: 22443507)
Neuron. 2019 Oct 23;104(2):305-321.e8. (PMID: 31474508)
Grant Information:
PP00P3_170672 Switzerland SNSF_ Swiss National Science Foundation; EN 1240/2-1 Deutsche Forschungsgemeinschaft; Starting Grant 803870 International ERC_ European Research Council; SFB 1134,TP03 Deutsche Forschungsgemeinschaft; PN 458054460 Deutsche Forschungsgemeinschaft; T32 GM140223 United States GM NIGMS NIH HHS; R01MH126960 United States NH NIH HHS; T32GM140223 Interdepartmental Training in Pharmacological Sciences; R01 MH126960 United States MH NIMH NIH HHS
Contributed Indexing:
Keywords: AIS plasticity; ankryin-G; axon initial segment; live imaging; mouse; neuroscience; node of Ranvier
Molecular Sequence:
Dryad 10.5061/dryad.n02v6wx76
Substance Nomenclature:
0 (Ankyrins)
147336-22-9 (Green Fluorescent Proteins)
0 (Ank3 protein, mouse)
0 (Recombinant Fusion Proteins)
147336-22-9 (Green Fluorescent Proteins)
0 (Ank3 protein, mouse)
0 (Recombinant Fusion Proteins)
Entry Date(s):
Date Created: 20250203 Date Completed: 20250505 Latest Revision: 20250913
Update Code:
20260130
PubMed Central ID:
PMC11790247
DOI:
10.7554/eLife.87078
PMID:
39898808
Database:
MEDLINE
*Further Information*
*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.
(© 2023, Thome et al.)*
*CT, JJ, SK, CA, ED, SS, KB, MB, NL, JR, NS, MH, DG, CB, JG, LM, KB, CS, VB, PJ, ME No competing interests declared*