Quantitative Assessment of Mesoscale Cellular Order and Organization in the Mouse Hippocampus

The hippocampus is characterized by a stereotypical macroscopic structure, where the nuclei are densely and heterogeneously packed among different subregions of the hippocampus. Despite the fact that tissue-specific cellular organization has been implicated in neural function, it has been technically challenging to quantitatively analyze mesoscopic cellular organization in the hippocampus due to its high cellular density. To overcome this technical hurdle, we developed Computational Biophysical Histomorphometry Software (CBHS), an automated image-analysis pipeline aimed at quantifying nuclear shape and the order of the cellular ensemble in high-density areas. When applied to the subfields of the hippocampus, we found that denser regions, most notably the dentate gyrus, were the most positionally, but least orientationally ordered. Nuclear shape exhibited a dependence on the local environment in a packing-dependent manner. This association was cell-type specific, with neurons, but not astrocytes displaying nuclear shape that varied with neighbor proximity, although astrocytes demonstrated greater intrinsic shape variance. The results reveal the presence of reproducible mesoscale cell packing order in hippocampal tissue, and are consistent with a nucleus-driven mechanical coupling between neighboring cells. The present study provides a quantitative framework with which to understand mesoscopic tissue organization, thus enabling the formulation of testable hypotheses for future investigation.