Abstract: TH-PO0547
Analysis of Renal Tubular Cell Turnover Using Doxycycline-Inducible Histone 2B-GFP Mice
Session Information
- Development, Stem Cells, and Regenerative Medicine
November 06, 2025 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 600 Development, Stem Cells, and Regenerative Medicine
Authors
- Takahashi, Shunsuke, Department of Nephrology and Hypertension, Saitama Medical Center, Saitama Medical University, Kawagoe, Saitama, Japan
- Okada, Mari, Department of Nephrology and Hypertension, Saitama Medical Center, Saitama Medical University, Kawagoe, Saitama, Japan
- Takayanagi, Kaori, Department of Nephrology and Hypertension, Saitama Medical Center, Saitama Medical University, Kawagoe, Saitama, Japan
- Hiromura, Keiju, Department of Nephrology and Rheumatology, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan
- Maeshima, Akito, Department of Nephrology and Hypertension, Saitama Medical Center, Saitama Medical University, Kawagoe, Saitama, Japan
Background
Renal tubular epithelial cells possess a remarkable ability to proliferate and regenerate following injury, ultimately differentiating into mature tubular cells. However, the mechanisms underlying their turnover under physiological conditions remain poorly understood. In this study, we investigated the dynamics of tubular cell turnover using TetOP-Histone 2B-GFP (TetOP-H2B-GFP) transgenic mice, in which proliferating cells can be temporally labeled with GFP under doxycycline (DOX) control.
Methods
Male TetOP-H2B-GFP mice aged 8 weeks, 12 months, and 18 months received DOX for various durations (8 hours, 24 hours, 72 hours, 1 week, 2 weeks, and 3 weeks). After defined chase periods, kidneys were harvested for histological analysis. Co-immunostaining with segment–specific nephron markers was performed to assess the localization and quantify GFP-positive cells in the normal kidney.
Results
GFP-positive tubular cells were observed following DOX administration, most of which were positive for AQP1 and negative for THP, NCC, and AQP2, indicating a proximal tubular origin. The number of GFP-positive cells increased proportionally with DOX exposure, with approximately 30% of cortical tubular cells labeled after 1 week of DOX treatment. During the chase period, the number of GFP-positive cells gradually decreased, reaching approximately 2% of cortical cells after 12 weeks of chase. Shortly after 8 hours of DOX administration, GFP-positive cells were observed as single cells, whereas after one week of chase, they were often found in pairs or triplets, suggesting cell division. In 18-month-old, aged mice, the number of GFP-positive cells was significantly reduced compared to that in 8-week-old mice.
Conclusion
These findings suggest that proximal tubular epithelial cells in the healthy kidney undergo continuous slow turnover, potentially contributing to the maintenance of renal tubular homeostasis.