Abstract
Quantifying the distribution of cells in every brain region is fundamental to attaining a comprehensive census of distinct neuronal and glial types. Until recently, estimating neuron numbers involved time-consuming procedures that were practically limited to stereological sampling. Progress in open-source image recognition software, growth in computing power, and unprecedented neuroinformatics developments now offer the potentially paradigm-shifting alternative of comprehensive cell-by-cell analysis in an entire brain region. The Allen Brain Atlas provides free digital access to complete series of raw Nissl-stained histological section images along with regional delineations. Automated cell segmentation of these data enables reliable and reproducible high-throughput quantification of regional variations in cell count, density, size, and shape at whole-system scale. While this strategy is directly applicable to any regions of the mouse brain, we first deploy it here on the closed-loop circuit of the hippocampal formation: the medial and lateral entorhinal cortices; dentate gyrus (DG); areas Cornu Ammonis 3 (CA3), CA2, and CA1; and dorsal and ventral subiculum. Using two independent image processing pipelines and the adult mouse reference atlas, we report the first cellular-level soma segmentation in every sub-region and non-principal layer of the left hippocampal formation through the full rostral-caudal extent. It is important to note that our techniques excluded the layers with the largest number of cells, DG granular and CA pyramidal, due to dense packing. The numerical estimates for the remaining layers are corroborated by traditional stereological sampling on a data subset and well match sparse published reports.
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Acknowledgements
We are grateful to Drs. Diek W. Wheeler (from the authors’ lab) and Christopher L. Rees (Harvard University) for critical discussions, and to Dr. Lydia Ng (Allen Institute for Brain Science) for sharing technical details about image scaling and resolution in the Allen Reference Atlas. This work was supported in part by grants R01 NS39600 and U01 MH114829 from the National Institutes of Health.
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Attili, S.M., Silva, M.F.M., Nguyen, Tv. et al. Cell numbers, distribution, shape, and regional variation throughout the murine hippocampal formation from the adult brain Allen Reference Atlas. Brain Struct Funct 224, 2883–2897 (2019). https://doi.org/10.1007/s00429-019-01940-7
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DOI: https://doi.org/10.1007/s00429-019-01940-7