Kidney Week

Abstract: TH-OR83

Cellular-Level Transcriptomics and Three-Dimensional Imaging for Recruited Renin Cells

Session Information

• Mechanisms of Hypertension and Cardiorenal Disease: From the Vasculature to the Gut
  November 02, 2023 | Location: Room 108, Pennsylvania Convention Center
  Abstract Time: 04:48 PM - 04:57 PM

Category: Hypertension and CVD

• 1601 Hypertension and CVD: Basic

Authors

• Yamaguchi, Hiroki, University of Virginia School of Medicine, Charlottesville, Virginia, United States

Hiroki Yamaguchi, MD, PhD

• Yamaguchi, Manako, University of Virginia School of Medicine, Charlottesville, Virginia, United States

Manako Yamaguchi,

• Watanabe, Hirofumi, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan

Hirofumi Watanabe,

• Medrano, Silvia, University of Virginia School of Medicine, Charlottesville, Virginia, United States

Silvia Medrano,

• Sequeira Lopez, Maria Luisa S., University of Virginia School of Medicine, Charlottesville, Virginia, United States

Maria Luisa S. Sequeira Lopez,

• Gomez, Roberto Ariel, University of Virginia School of Medicine, Charlottesville, Virginia, United States

Roberto Ariel Gomez,

Background

Renin synthesis is tightly regulated by the ability of juxtaglomerular cells (JG) to sense arterial pressure signals. Once severe hypotension occurs in adult animals, JGs promote renin synthesis first. Subsequently, a subset of vascular smooth muscle cells (VSMC) derived from renin progenitors turn on renin expression to restore homeostasis: this phenomenon is termed recruitment. However, identifying the mechanisms of the recruitment of renin lineage cells has been challenging due to their rarity and structural complexity. In this study, we investigated the transcriptome profiles and expression patterns of individual recruited renin cells by single-cell RNA-seq and three-dimensional (3D) imaging.

Methods

Since forkhead box protein D1 (FoxD1) expressing stromal cells are the progenitors for JGs and VSMCs of the renal arteriole, we generated FoxD1-GC; R26R^TdTomato mice, where all FoxD1⁺ descendants express tdTomato reporter. We applied a surgical model of aortic coarctation (AoCo) between the base at the renal arteries of the left kidney (LK) and right kidneys (RK) that received low and high perfusion pressure, respectively. We FACS sorted tdTomato⁺ cells from the LK and RK cortices and performed single-cell RNA-seq to analyze the differentially expressed genes (DEG) and the gene ontologies between LK and RK. To visualize renin-expressing cells in 3D, we applied the clear, unobstructed brain/body imaging cocktails (CUBIC) protocol for clearing kidneys of the Ren1^c-tdTomato mice that underwent AoCo surgery. We used Zeiss Lightsheet7 for 3D imaging.

Results

By single-cell RNA-seq, FoxD1⁺ cells were clustered into JG, VSMC, and other cell types. DEG analysis in LK compared to RK showed that Ren1 was the upregulated DEG in JG and VSMC. In VSMC, gene ontology analysis showed the upregulation of the cellular response to laminar fluid shear stress pathway in LK, suggesting that extracellular force changes affect the gene expression. By CUBIC protocol, renin-expressing cells in the Ren1^c-tdTomato mouse kidneys could be visualized in 3D. In LK, we could identify recruited VSMCs forming a stripped and ring pattern along with afferent arterioles, some extending close to or beyond the bifurcation point.

Conclusion

Severe hypotension causes the widespread recruitment of renin lineage VSMCs in the renal arteriole.

Funding

• Other NIH Support