A Simple Method for 3D Analysis of Immunolabeled Axonal Tracts in a Transparent Nervous System
Clearing techniques have been developed to transparentize mouse brains, thereby preserving 3D structure, but their complexity has limited their use. Here, we show that immunolabeling of axonal tracts followed by optical clearing with solvents (3DISCO) and light-sheet microscopy reveals brain connect...
Main Authors: | , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Elsevier
2014-11-01
|
Series: | Cell Reports |
Online Access: | http://www.sciencedirect.com/science/article/pii/S2211124714009061 |
id |
doaj-79c5b43caa5b408187a1c34a93ac2398 |
---|---|
record_format |
Article |
spelling |
doaj-79c5b43caa5b408187a1c34a93ac23982020-11-25T02:07:58ZengElsevierCell Reports2211-12472014-11-01941191120110.1016/j.celrep.2014.10.037A Simple Method for 3D Analysis of Immunolabeled Axonal Tracts in a Transparent Nervous SystemMorgane Belle0David Godefroy1Chloé Dominici2Céline Heitz-Marchaland3Pavol Zelina4Farida Hellal5Frank Bradke6Alain Chédotal7Sorbonne Universités, UPMC Université Paris 06, UMRS 968, Institut de la Vision, Paris 75012, FranceSorbonne Universités, UPMC Université Paris 06, UMRS 968, Institut de la Vision, Paris 75012, FranceSorbonne Universités, UPMC Université Paris 06, UMRS 968, Institut de la Vision, Paris 75012, FranceSorbonne Universités, UPMC Université Paris 06, UMRS 968, Institut de la Vision, Paris 75012, FranceSorbonne Universités, UPMC Université Paris 06, UMRS 968, Institut de la Vision, Paris 75012, FranceDeutsches Zentrum für Neurodegenerative Erkrankungen (DZNE), Axon Growth and Regeneration, Ludwig-Erhard-Allee 2, 53175 Bonn, GermanyDeutsches Zentrum für Neurodegenerative Erkrankungen (DZNE), Axon Growth and Regeneration, Ludwig-Erhard-Allee 2, 53175 Bonn, GermanySorbonne Universités, UPMC Université Paris 06, UMRS 968, Institut de la Vision, Paris 75012, FranceClearing techniques have been developed to transparentize mouse brains, thereby preserving 3D structure, but their complexity has limited their use. Here, we show that immunolabeling of axonal tracts followed by optical clearing with solvents (3DISCO) and light-sheet microscopy reveals brain connectivity in mouse embryos and postnatal brains. We show that the Robo3 receptor is selectively expressed by medial habenula axons forming the fasciculus retroflexus (FR) and analyzed the development of this commissural tract in mutants of the Slit/Robo and DCC/Netrin pathways. Netrin-1 and DCC are required to attract FR axons to the midline, but the two mutants exhibit specific and heterogeneous axon guidance defects. Moreover, floor-plate-specific deletion of Slit ligands with a conditional Slit2 allele perturbs not only midline crossing by FR axons but also their anteroposterior distribution. In conclusion, this method represents a unique and powerful imaging tool to study axonal connectivity in mutant mice.http://www.sciencedirect.com/science/article/pii/S2211124714009061 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Morgane Belle David Godefroy Chloé Dominici Céline Heitz-Marchaland Pavol Zelina Farida Hellal Frank Bradke Alain Chédotal |
spellingShingle |
Morgane Belle David Godefroy Chloé Dominici Céline Heitz-Marchaland Pavol Zelina Farida Hellal Frank Bradke Alain Chédotal A Simple Method for 3D Analysis of Immunolabeled Axonal Tracts in a Transparent Nervous System Cell Reports |
author_facet |
Morgane Belle David Godefroy Chloé Dominici Céline Heitz-Marchaland Pavol Zelina Farida Hellal Frank Bradke Alain Chédotal |
author_sort |
Morgane Belle |
title |
A Simple Method for 3D Analysis of Immunolabeled Axonal Tracts in a Transparent Nervous System |
title_short |
A Simple Method for 3D Analysis of Immunolabeled Axonal Tracts in a Transparent Nervous System |
title_full |
A Simple Method for 3D Analysis of Immunolabeled Axonal Tracts in a Transparent Nervous System |
title_fullStr |
A Simple Method for 3D Analysis of Immunolabeled Axonal Tracts in a Transparent Nervous System |
title_full_unstemmed |
A Simple Method for 3D Analysis of Immunolabeled Axonal Tracts in a Transparent Nervous System |
title_sort |
simple method for 3d analysis of immunolabeled axonal tracts in a transparent nervous system |
publisher |
Elsevier |
series |
Cell Reports |
issn |
2211-1247 |
publishDate |
2014-11-01 |
description |
Clearing techniques have been developed to transparentize mouse brains, thereby preserving 3D structure, but their complexity has limited their use. Here, we show that immunolabeling of axonal tracts followed by optical clearing with solvents (3DISCO) and light-sheet microscopy reveals brain connectivity in mouse embryos and postnatal brains. We show that the Robo3 receptor is selectively expressed by medial habenula axons forming the fasciculus retroflexus (FR) and analyzed the development of this commissural tract in mutants of the Slit/Robo and DCC/Netrin pathways. Netrin-1 and DCC are required to attract FR axons to the midline, but the two mutants exhibit specific and heterogeneous axon guidance defects. Moreover, floor-plate-specific deletion of Slit ligands with a conditional Slit2 allele perturbs not only midline crossing by FR axons but also their anteroposterior distribution. In conclusion, this method represents a unique and powerful imaging tool to study axonal connectivity in mutant mice. |
url |
http://www.sciencedirect.com/science/article/pii/S2211124714009061 |
work_keys_str_mv |
AT morganebelle asimplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT davidgodefroy asimplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT chloedominici asimplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT celineheitzmarchaland asimplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT pavolzelina asimplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT faridahellal asimplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT frankbradke asimplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT alainchedotal asimplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT morganebelle simplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT davidgodefroy simplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT chloedominici simplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT celineheitzmarchaland simplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT pavolzelina simplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT faridahellal simplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT frankbradke simplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem AT alainchedotal simplemethodfor3danalysisofimmunolabeledaxonaltractsinatransparentnervoussystem |
_version_ |
1724928462581399552 |