Three-dimensional (3D) mural cell morphology is heterogeneous and coupled to vessel geometry, however, measurements from two-dimensional (2D) maximum intensity projections (MIP) obscure overlapping processes and cell-vessel contacts. Accordingly, we developed Mural-VISTA, a semi-automated Python workflow for mural cell-vessel interaction and single-cell topo-morphology analysis of reconstructed surface meshes. This workflow integrates mesh pretreatment, interactive centerline extraction, hierarchical segmentation of cell soma, main axis and secondary processes (branches), and extraction of 36 multiscale (cell process segment level, process level, and whole cell level) topo-morphological and vessel-referenced metrics. Mural-VISTA identified morphological changes in pericytes and vascular smooth muscle cells (vSMCs) with altered RhoA activity. Constitutive active RhoA (RhoA CA) over-expression reduced branch complexity and increased process alignment in both cell types, while increased whole-cell and branch solidity only in vSMCs. Dominant negative RhoA (RhoA DN) over-expression increased branch abundance and reduced branch solidity in pericytes but not vSMCs, suggesting cell-type specific effect of reduced RhoA activity. In conclusion, Mural-VISTA enables quantitative 3D profiling of mural cell architecture and its spatial relationship with the vessel.
Zeng, H., Hu, M., Phng, L.-K., Matsunaga, Y. T.
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