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.

微血管研究领域的新兴方向聚焦于神经与血管模式构建之间的关联。然而，成年组织中血管与神经生长之间的功能依赖性仍未得到充分探究。
本研究旨在明确成年微血管生长时序过程中，血管与神经网络之间的时空协同关系。
研究采集了成年雄性Wistar大鼠的肠系膜组织，分别于刺激前，以及肥大细胞脱颗粒诱导血管生成后的第2、10和30天取材。
对组织进行免疫标记，以检测PECAM（内皮细胞标记物）与Ⅲ类β微管蛋白（周围神经标记物）。
研究人员按血管类别对神经血管对齐程度开展定量分析：包括小动脉（直径>20μm）、毛细血管前动脉（直径10~20μm）、毛细血管后微静脉（直径10~20μm）、微静脉（直径>20μm）、毛细血管（直径<10μm）以及毛细血管芽。
在第2天和第10天，毛细血管前动脉、毛细血管、毛细血管后微静脉以及微静脉的神经血管对齐程度相较于未刺激组出现下降。
该下降程度与各血管类别的血管密度升高呈负相关。
至第30天时，对齐程度要么恢复至未刺激组水平，要么相较于第10天有所提升。
上述结果表明，神经血管对齐程度在微血管网络生长完成后形成，并存在于小动脉、微静脉乃至毛细血管中。