Department of Pathology and Medical Biology, Medical Biology Section, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.
Department of Critical Care, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.
Am J Physiol Renal Physiol. 2023 Sep 1;325(3):F299-F316. doi: 10.1152/ajprenal.00005.2023. Epub 2023 Jul 6.
Endothelial cells in blood vessels in the kidney exert different functions depending on the (micro)vascular bed they are located in. The present study aimed to investigate microRNA and mRNA transcription patterns that underlie these differences. We zoomed in on microvascular compartments in the mouse renal cortex by laser microdissecting the microvessels prior to small RNA- and RNA-sequencing analyses. By these means, we characterized microRNA and mRNA transcription profiles of arterioles, glomeruli, peritubular capillaries, and postcapillary venules. Quantitative RT-PCR, in situ hybridization, and immunohistochemistry were used to validate sequencing results. Unique microRNA and mRNA transcription profiles were found in all microvascular compartments, with dedicated marker microRNAs and mRNAs showing enriched transcription in a single microvascular compartment. In situ hybridization validated the localization of microRNAs mmu-miR-140-3p in arterioles, mmu-miR-322-3p in glomeruli, and mmu-miR-451a in postcapillary venules. Immunohistochemical staining showed that von Willebrand factor protein was mainly expressed in arterioles and postcapillary venules, whereas GABRB1 expression was enriched in glomeruli, and IGF1 was enriched in postcapillary venules. More than 550 compartment-specific microRNA-mRNA interaction pairs were identified that carry functional implications for microvascular behavior. In conclusion, our study identified unique microRNA and mRNA transcription patterns in microvascular compartments of the mouse kidney cortex that underlie microvascular heterogeneity. These patterns provide important molecular information for future studies into differential microvascular engagement in health and disease. Renal endothelial cells display a high level of heterogeneity depending on the (micro)vascular bed they reside in. The molecular basis contributing to these differences is poorly understood yet of high importance to increase understanding of microvascular engagement in the kidney in health and disease. This report describes m(icro)RNA expression profiles of microvascular beds in the mouse renal cortex and uncovers microvascular compartment-specific m(icro)RNAs and miRNA-mRNA pairs, thereby revealing important molecular mechanisms underlying renal microvascular heterogeneity.
肾脏血管中的内皮细胞根据其所在的（微血管）床而发挥不同的功能。本研究旨在探讨这些差异背后的 microRNA 和 mRNA 转录模式。我们通过在进行小 RNA 和 RNA 测序分析之前激光显微切割微血管，聚焦于小鼠肾皮质的微血管隔室。通过这些方法，我们对小动脉、肾小球、肾小管周围毛细血管和小静脉进行了 microRNA 和 mRNA 转录谱的特征描述。定量 RT-PCR、原位杂交和免疫组织化学用于验证测序结果。所有微血管隔室均具有独特的 microRNA 和 mRNA 转录谱，具有特定标记 microRNAs 和 mRNAs 的基因在单个微血管隔室中表现出丰富的转录。原位杂交验证了 microRNA mmu-miR-140-3p 在小动脉中的定位、mmu-miR-322-3p 在肾小球中的定位和 mmu-miR-451a 在小静脉中的定位。免疫组织化学染色显示，血管性血友病因子蛋白主要在小动脉和小静脉中表达，而 GABRB1 表达在肾小球中富集，IGF1 在小静脉中富集。鉴定出超过 550 个具有特定隔室的 microRNA-mRNA 相互作用对，这些相互作用对具有对微血管行为的功能意义。总之，本研究确定了小鼠肾皮质微血管隔室中独特的 microRNA 和 mRNA 转录模式，这些模式为未来研究健康和疾病状态下微血管的差异参与提供了重要的分子信息。肾脏内皮细胞根据其所在的（微血管）床表现出高度的异质性。导致这些差异的分子基础尚不清楚，但对于增加对肾脏微血管在健康和疾病中的参与的理解非常重要。本报告描述了小鼠肾皮质微血管床的 m(icro)RNA 表达谱，并揭示了微血管隔室特异性的 m(icro)RNAs 和 miRNA-mRNA 对，从而揭示了肾脏微血管异质性的重要分子机制。
