Abstract: FR-OR042
Observation of Renin Lineage Cell Migration Following Local Laser Damage by Longitudinal Intravital Multiphoton Microscopy
Session Information
- Development and Stem Cells
November 08, 2019 | Location: 152, Walter E. Washington Convention Center
Abstract Time: 04:42 PM - 04:54 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 500 Development, Stem Cells, and Regenerative Medicine
Authors
- Raith, Lisa, University Hospital CGC, Dresden University of Technology, Dresden, Germany
- Gerlach, Michael, TU Dresden, Dresden, Germany
- Sradnick, Jan, University Hospital CGC, Dresden University of Technology, Dresden, Germany
- Todorov, Vladimir T., University Hospital CGC, Dresden University of Technology, Dresden, Germany
- Hugo, Christian, University Hospital CGC, Dresden University of Technology, Dresden, Germany
Background
Several studies demonstrate intraglomerular migration of renin lineage cells (RLC) after antibody-induced damage of glomeruli. The aim of our study was to trigger and trace RLC migration patterns by non-systemic, spatially defined glomerular injury. Using an inducible transgenic RLC reporter mouse strain we were able to track cell migration by intravital 2-photon microscopy.
Methods
Renin lineage reporter mice expressing tdTomato (mRen-rtTAm2-LC1-tdT) were induced for 16 days. 24 hours before initial microscopy, abdominal body windows were implanted to allow repeated kidney imaging without further surgery. Single glomeruli were longitudinally examined six times (d0, d1, d2, d3, d6, d10) by 2-photon laser scanning microscope. Blood plasma was visualized by FITC dextran injection. Spatially defined damage was induced by intraglomerular scanning with a femtosecond pulsed 2-photon laser at high zoom factor, with a second observation 10 minutes after injury. Neighbouring non-damaged glomeruli in the same animal served as controls. Data was 3D analysed with Bitplane Imaris. Glomerular volume, total volume of intraglomerular tdTomato positive cells and maximal migration distance from the juxtaglomerular apparatus to tdTomato positive area were evaluated.
Results
RLC migration into the injured glomeruli could be observed as early as day 3 after damage. The total glomerular volume was reduced by injured glomerular over time. Compared to controls, intra-glomerular tdTomato positive cell volume continuously elevated on day 3, day 6 and day 10 (0.4 ±0.7%, 2.4 ±2.2% , 7.3 ± 7.8%). Maximal migration distance also progressively increased during observed times (day 3: 7.1 ± 6.7 µm, day 6: 21.3 ±11.9 µm and day 10: 26.8 ± 14.1 µm).
Conclusion
We were able to evaluate the temporal and spatial migration pattern of RLC into single glomeruli, starting as early as 3 days after injury. Moreover, we quantitatively assessed this process by 3D analysis. This new approach gives us the opportunity to characterize the migratory path of RLC after glomerular injury and evaluate the impact of local signalling due to spatially restricted damage.