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Studying connectivity and brain development with combinatorial Brainbow labels

Karine Loulier1, Katherine S. Matho1, Yann Le Franc2, Willy Supatto3, Raphaelle Barry1, Pierre Mahou3, Sio Hoi Ieng1, Stephane Fouquet1, Alain Chedotal1, Ryad B. Benosman1, Emmanuel Beaurepaire3, Xavier Morin4 and Jean Livet1*
  • 1 Institut de la Vision, INSERM U968, UPMC UMR_S 968, CNRS UMR 7210, France
  • 2 University of Antwerp, Belgium
  • 3 Ecole Polytechnique, France
  • 4 Ecole Normale Superieure, Institut de Biologie de l'ENS, INSERM U1024, CNRS UMR 8197, France

Neural circuits of the brain are highly complex cellular architectures that assemble through a series of elaborate developmental processes. Mapping these architectures and tracing back their developmental history at the individual cell level are essential tasks for deciphering the building principles of neural circuits. Monochrome markers used so far to single-out neural cells or clones in the brain are painstaking and necessitate a high degree of sparseness. Brainbow multicolor labeling strategies solve this problem by contrasting neighboring cells of a tissue from one another with combinations of spectrally distinct fluorescent proteins (FPs). We present an overview of this approach, highlighting new tools and methods that we have developed for expressing and analyzing multicolor labels in the nervous system of developing and adult vertebrate models. We show how dense Brainbow labeling can be used to map connections in specific brain stem circuits presenting large axons and synapses. In parallel, we present new strategies for semi-sparse multicolor labeling which open the way for multiplex tracing of neural anatomy and cell lineage in brain areas presenting smaller neural component such as the mouse forebrain. We demonstrate new imaging options suitable for visualizing Brainbow markers in these two contexts. Finally, we present methods to quantify FP combinations expressed by labeled cells for the purpose of deciphering their lineage relationships. These methodologies provide new ways to delineate cytoarchitecture, resolve spatially intermixed neural clones and specify neural and glial cell lineage.

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Keywords: Brainbow, lineage, multicolor marker, tissue reconstruction, central nervous system development

Conference: Imaging the brain at different scales: How to integrate multi-scale structural information?, Antwerp, Belgium, 2 Sep - 6 Sep, 2013.

Presentation Type: Lecture

Topic: Handling of light and magnetic resonance microscopy data sets

Citation: Loulier K, Matho KS, Le Franc Y, Supatto W, Barry R, Mahou P, Ieng S, Fouquet S, Chedotal A, Benosman RB, Beaurepaire E, Morin X and Livet J (2013). Studying connectivity and brain development with combinatorial Brainbow labels. Front. Neuroinform. Conference Abstract: Imaging the brain at different scales: How to integrate multi-scale structural information?. doi: 10.3389/conf.fninf.2013.10.00010

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Received: 21 Aug 2013; Published Online: 30 Aug 2013.

* Correspondence: Dr. Jean Livet, Institut de la Vision, INSERM U968, UPMC UMR_S 968, CNRS UMR 7210, Paris, France, jean.livet@inserm.fr