Quantitative neuroanatomy of all Purkinje cells with light sheet microscopy and high-throughput image analysis

Year: 2015

Authors: Silvestri L., Paciscopi M., Soda P., Biamonte F., Iannello G., Frasconi P., Pavone F. S.

Autors Affiliation: Istituto Nazionale di Ottica (INO-CNR), Sesto Fiorentino (FI), Italy;
Laboratorio Europeo di Spettroscopie Nonlineare (LENS) Sesto Fiorentino (FI), Italy;
Univ Florence, Dept Informat Engn, Florence, Italy;
University Campus Bio-Medico, Dept Engn, Rome, Italy;
Catholic Univ Sacred Heart A Gemelli, Inst Histol & Embryol, Rome, Italy
Univ Florence, Dept Phys & Astron, Sesto Fiorentino, Italy
[ 7 ] Int Ctr Computat Neurophoton, Sesto Fiorentino, Italy

Abstract: Characterizing the cytoarchitecture of mammalian central nervous system on a brainwide scale is becoming a compelling need in neuroscience. For example, realistic modeling of brain activity requires the definition of quantitative features of large neuronal populations in the whole brain. Quantitative anatomical maps will also be crucial to classify the cytoarchtitectonic abnormalities associated with neuronal pathologies in a high reproducible and reliable manner. In this paper, we apply recent advances in optical microscopy and image analysis to characterize the spatial distribution of Purkinje cells (PCs) across the whole cerebellum. Light sheet microscopy was used to image with micron-scale resolution a fixed and cleared cerebellum of an L7-GFP transgenic mouse, in which all PCs are fluorescently labeled. A fast and scalable algorithm for fully automated cell identification was applied on the image to extract the position of all the fluorescent PCs. This vectorized representation of the cell population allows a thorough characterization of the complex three-dimensional distribution of the neurons, highlighting the presence of gaps inside the lamellar organization of PCs, whose density is believed to play a significant role in autism spectrum disorders. Furthermore, clustering analysis of the localized somata permits dividing the whole cerebellum in groups of PCs with high spatial correlation, suggesting new possibilities of anatomical partition. The quantitative approach presented here can be extended to study the distribution of different types of cell in many brain regions and across the whole encephalon, providing a robust base for building realistic computational models of the brain, and for unbiased morphological tissue screening in presence of pathologies and/or drug treatments.


Volume: 9      Pages from: 68  to: 68

More Information: The research leading to these results has received funding from the European Union Seventh Framework Program (FP7/2007-2013) under grant agreements no. 604102 (Human Brain Project) and no. 284464 (LASERLAB-EUROPE). The research has also been supported by the Italian Ministry for Education, University and Research in the framework of the Flagship Project Nano MAX, by \”Ente Cassa di Risparmio di Firenze\” (private foundation).
KeyWords: quantitative neuroanatomy; Purkinje cells; cerebellum; light sheet microscopy; image analysis
DOI: 10.3389/fnana.2015.00068

Citations: 31
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