Supplementary Material for: Spatiotemporal Distribution of Neurovascular Alignment in Remodeling Adult Rat Mesentery Microvascular Networks

An emerging area of microvascular research focuses on the links between neural and vascular patterning. However, the functional dependence between vascular and neural growth in adult tissues remains underinvestigated. The objective of this study was to determine the spatial and temporal coordination between vascular and neural networks over a time course of adult microvascular growth. Mesentery tissues from adult male Wistar rats were harvested prior to stimulation, and 2, 10 and 30 days after angiogenesis stimulated by mast cell degranulation. Tissues were immunolabeled for PECAM (endothelial cell marker) and class III β-tubulin (peripheral nerve marker). Neurovascular alignment was quantified per vessel category: arterioles (>20 µm), pre-capillary arterioles (10–20 µm), post-capillary venules (10–20 µm), venules (>20 µm), capillaries (<10 µm) and capillary sprouts. Neurovascular alignment along pre-capillary arterioles, capillaries, post-capillary venules and venules was decreased compared to unstimulated levels on days 2 and 10. These decreases inversely correlated with increases in vessel density per vessel category. By day 30, alignment either returned to unstimulated levels or was increased compared to day 10. These results suggest that neurovascular alignment arises after microvascular network growth and is present along arterioles, venules and even capillaries.