A Simple Method for 3D Analysis of Immunolabeled Axonal Tracts in a Transparent Nervous System

被引：147

作者：

Belle, Morgane ^{[1
,2
,3
]}

Godefroy, David ^{[1
,2
,3
]}

Dominici, Chloe ^{[1
,2
,3
]}

Heitz-Marchaland, Celine ^{[1
,2
,3
]}

Zelina, Pavol ^{[1
,2
,3
]}

Hellal, Farida ^{[4
]}

Bradke, Frank ^{[4
]}

Chedotal, Alain ^{[1
,2
,3
]}

机构：

[1] Sorbonne Univ, Univ Paris 06, UMRS 968, Inst Vis, F-75012 Paris, France

[2] INSERM, Inst Vis, UMRS 968, F-75012 Paris, France

[3] CNRS, UMR 7210, F-75012 Paris, France

[4] Deutsch Zentrum Neurodegenerat Erkrankungen DZNE, D-53175 Bonn, Germany

来源：

CELL REPORTS | 2014年 / 9卷 / 04期

关键词：

GROWTH CONE GUIDANCE; SEMAPHORIN; 3F; MOUSE; HABENULA; SLIT; NEURONS; GENES; RECONSTRUCTION; RIG-1/ROBO3; EXPRESSION;

D O I：

10.1016/j.celrep.2014.10.037

中图分类号：

Q2 [细胞生物学];

学科分类号：

071013 [干细胞生物学];

摘要：

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.

引用

页码：1191 / 1201

页数：11

共 57 条

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Identification of the Zebrafish Ventral Habenula As a Homolog of the Mammalian Lateral Habenula [J].