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Neuroimage. 2016 Sep;138:64-75. doi: 10.1016/j.neuroimage.2016.05.050. Epub 2016 May 21.

Column-like Ca(2+) clusters in the mouse neonatal neocortex revealed by three-dimensional two-photon Ca(2+) imaging in vivo.

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Hans-Berger Department of Neurology, University Hospital Jena, D-07747 Jena, Germany.
Institute of Biomedical Engineering and Informatics, Technical University Ilmenau, D-98693 Ilmenau, Germany.
Hans-Berger Department of Neurology, University Hospital Jena, D-07747 Jena, Germany. Electronic address:


Neuronal network activity in the developing brain is generated in a discontinuous manner. In the visual cortex during the period of physiological blindness of immaturity, this activity mainly comprises retinally triggered spindle bursts or Ca(2+) clusters thought to contribute to the activity-dependent construction of cortical circuits. In spite of potentially important developmental functions, the spatial structure of these activity patterns remains largely unclear. In order to address this issue, we here used three-dimensional two-photon Ca(2+) imaging in the visual cortex of neonatal mice at postnatal days (P) 3-4 in vivo. Large-scale voxel imaging covering a cortical depth of 200μm revealed that Ca(2+) clusters, identified as spindle bursts in simultaneous extracellular recordings, recruit cortical glutamatergic neurons of the upper cortical plate (CP) in a column-like manner. Specifically, the majority of Ca(2+) clusters exhibit prominent horizontal confinement and high intra-cluster density of activation involving the entire depth of the upper CP. Moreover, using simultaneous Ca(2+) imaging from hundreds of neurons at single-cellular resolution, we demonstrate that the degree of neuronal co-activation within Ca(2+) clusters displays substantial heterogeneity. We further provide evidence that co-activated cells within Ca(2+) clusters are spatially distributed in a non-stochastic manner. In summary, our data support the conclusion that dense coding in the form of column-like Ca(2+) clusters is a characteristic property of network activity in the developing visual neocortex. Such knowledge is expected to be relevant for a refined understanding of how specific spatiotemporal characteristics of early network activity instruct the development of cortical circuits.


Calcium imaging; Development; Network activity; Two-photon microscopy; Visual cortex

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