WM-CLICK, a new method for 3d detection, representation and analysis of dividing cells in the whole brain

Lazutkin, A, Shuvaev, S, Kiryanov, R, Anokhin, K, Koulakov, A, Enikolopov, G (June 2018) WM-CLICK, a new method for 3d detection, representation and analysis of dividing cells in the whole brain. Opera Medica et Physiologica, 4. p. 20. ISSN 2500-2287

DOI: 10.20388/omp2018.00s1.001


Recent years are marked by the active development of methods of three-dimensional optical visualization of the whole brain. Efficient imaging, quantification, and analysis carried out in 3D may reveal dynamics and hidden patterns of processes not recognized on conventional flat sections. This holds especially true for the study of neurogenesis both in the developing and adult nervous system where neural stem and progenitor cells divide in restricted regions and migrate along intricate trajectories to reach distant areas of the brain. We have recently developed a new histological technique for 3D imaging of proliferating cells in the whole brain of developing and adult mice, based on labeling the dividing cells with 5-ethynyl-2’-deoxyuridine (EdU) and detecting them with fluorescent azide using whole-mount click-reaction (WM-CLICK). We also developed novel methods for automatic volume registration, counting and morphing of 3D images. We applied these techniques, combined with scanning confocal microscopy, light-sheet microscopy and two-photon tomography for visualizing patterns of cell division in the mouse brain. We observed three proliferation/migration streams in the subventricular zone (SVZ) of adult mouse brain – dorsolateral, dorsomedial and ventral branches which merge together into a common rostral migration stream and then traced the dynamics of their formation. We also used WM-CLICK and our new mathematic algorithms to compare the 3D patterns of cell division in control and memantine-treated adult brain. Several areas with strong effects of memantine on cell division were revealed. The higher densities of EdU+cells were found in the CAs and DG regions of hippocampus, subcallosal zone, olfactory bulb, and piriform cortex. We also observed profound changes in the patterns of cell division at the perinatal age in two brain regions – SVZ, where streams are gradually formed during development, and cerebellum, where we observed heterochrony in proliferation intensity in the lateral and medial parts. The resulting data can be presented as pseudo time-lapse movie, thus allowing 4D representation of the dynamics of developmental changes in cell division and neurogenesis. This study was supported by grant 11.G34.31.0071 from Russian Ministry of Education and Science and by grants of RFBR no. 15-29-01305 and 17-29-01037 and RSCF no. 16-15-00294 and 17-15-01426

Item Type: Paper
Subjects: organs, tissues, organelles, cell types and functions > cell types and functions > cell functions > cell division
CSHL Authors:
Communities: CSHL labs > Koulakov lab
SWORD Depositor: CSHL Elements
Depositing User: CSHL Elements
Date: 1 June 2018
Date Deposited: 06 May 2021 15:50
Last Modified: 24 Apr 2024 15:10
URI: https://repository.cshl.edu/id/eprint/40008

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