Abstract: SA-PO104
Intravital Imaging of Single Collecting Ducts Micro-Perfused with Uropathogenic Escherichia coli Demonstrates Phagocytosis by Intercalated Cells
Session Information
- AKI: Mechanisms - Primary Injury and Repair - II
November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- Schwaderer, Andrew L., Indiana University, Zionsville, Indiana, United States
- Hato, Takashi, Indiana University, Zionsville, Indiana, United States
- Hains, David S., Indiana University, Zionsville, Indiana, United States
- Saxena, Vijay, Indiana University, Zionsville, Indiana, United States
- Arregui, Sam W., IU School of Medicine, Indianapolis, Indiana, United States
Background
Renal epithelial cells have a different lineage and historically were considered to have different functions than myeloid derived macrophages. However some epithelial cells, such as renal intercalated cells have been recently demonstrated to have distinct immune functions.
Methods
VATPaseB1-cre transgenic mice were crossed to tdTomato-loxp homozygous mice which results in red fluorescence of intercalated cells. Under isoflurane anesthesia a kidney was surgically exposed and a single collecting duct was cannulated with a 5 micron needle. The tubule was perfused with green fluorescent protein (GFP) expressing uropathogenic E.coli (UPEC) strain CFT 073 or E.coli coated bioparticles and imaged with Leica TCS SP8 (upright high-speed multiphoton and confocal imaging system).
Results
Single collecting ducts could be cannulated (Figure 1A). Micro-perfusion with UPEC could be confirmed by visualization GFP expressing bacteria flowing through a tubular structure (Figure 1B). Bacteria were internalized selectively in red fluorescent expressing intercalated cells (Figure 1C-D). Ecoli coated bioparticles were also selectively internalized by intercalated indicating an intercalated cell activated process not just bacterial invasion.
Conclusion
We demonstrate that micro-perfusion of single collecting ducts can be accomplished in vivo and imaged using intravital microscopy. Renal intercalated cells phagocytize bacteria similar to myeloid immune cells such as macrophages even though intercalated cells are renal epithelial cells.
Funding
- NIDDK Support