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Published on: October 18, 2016
Three-Dimensional Visualization With Tissue Clearing Uncovers Dynamic Alterations of Renal Resident Mononuclear
Kipyo Kim1, Yun-Gyeong Kim2, Su Woong Jung2
1Division of Nephrology and Hypertension, Department of Internal Medicine, Inha University School of Medicine, Incheon, South Korea.
Insights
Optical tissue clearing with advanced microscopy reveals dynamic changes in immune cells during acute kidney injury (AKI). This study shows NLRP3 inhibition improves kidney function by reducing immune cell activation in AKI.
Area of Science:
- Nephrology
- Immunology
- Microscopy
Background:
- Traditional histology struggles to capture dynamic immune cell changes in acute kidney injury (AKI).
- Optical tissue clearing combined with multiphoton microscopy (MPM) or light sheet fluorescence microscopy (LSFM) offers advanced deep tissue visualization.
- These techniques enhance understanding of immune cell dynamics during tissue injury and repair.
Purpose of the Study:
- To investigate morphological and functional alterations of renal mononuclear phagocytes (MNPs) in lipopolysaccharide (LPS)-induced AKI.
- To evaluate the effect of the NLRP3 inhibitor MCC950 on CD11c+ cell activation in LPS-induced AKI.
- To utilize renal clearing and advanced microscopy for 3D visualization of immune cell changes in AKI.
Main Methods:
- Utilized CD11c-YFP mice for studying renal mononuclear phagocytes (MNPs).
- Employed optical tissue clearing techniques for whole mouse kidney imaging.
- Performed deep tissue imaging using light sheet fluorescence microscopy (LSFM) and multiphoton microscopy (MPM).
- Assessed immune cell activation markers (MHC class II, CD86) and renal function.
Main Results:
- LPS-induced AKI significantly increased CD11c+ cell numbers and density, particularly in periglomerular and perivascular areas.
- High-resolution MPM revealed morphological changes in CD11c+ cells, including increased dendrites.
- Activated CD11c+ cells showed increased MHC class II and CD86 expression.
- Treatment with NLRP3 inhibitor MCC950 attenuated CD11c+ cell activation and improved renal function.
Conclusions:
- 3D visualization techniques (MPM, LSFM) combined with kidney clearing effectively reveal dynamic changes in renal MNPs during AKI.
- These dynamic immune cell alterations are directly linked to renal function.
- NLRP3 inhibition represents a potential therapeutic strategy for AKI by modulating immune cell activation.
Abstract:
Traditional histologic methods are limited in detecting dynamic changes in immune cells during acute kidney injury (AKI). Recently, optical tissue clearing combined with multiphoton microscopy (MPM) or light sheet fluorescence microscopy (LSFM) has become an emerging method for deep tissue evaluation and three-dimensional visualization. These new approaches have helped expand our understanding of tissue injury and repair processes, including tracing the changes in immune cells. We designed this study to investigate the morphological and functional alterations of renal mononuclear phagocytes (MNPs) in lipopolysaccharide (LPS)-induced AKI using renal clearing in CD11c-YFP mice. We also evaluated the effect of the NLRP3 inhibitor MCC950 to determine whether NLRP3 inhibition attenuates the activation of CD11c+ cells in an LPS-induced AKI model. Transverse sectioned whole mouse kidney imaging by LSFM showed that CD11c+ cells were mainly distributed in the cortex, especially the tubulointerstitial area. The number of CD11c+ cells was significantly more densely interspersed, particularly in periglomerular and perivascular lesions, in the saline-treated LPS-exposed kidney than in the control kidney. Deep imaging of the kidney cortex by MPM demonstrated an increased number of CD11c+ cells in the saline-treated LPS group compared with the control group. This quantitative alteration of CD11c+ cells in AKI was accompanied by morphological changes at high resolution, showing an increased number and level of dendrites. These morphological and behavioral changes in the saline-treated LPS group were accompanied by increased MHC class II and CD86 on CD11c-YFP+ cells. MCC950 attenuated the activation of CD11c+ cells after AKI and improved renal function. In conclusion, wide and deep three-dimensional visualization using MPM or LSFM combined with kidney clearing uncovers dynamic changes of renal MNPs, which are directly linked to renal function in AKI.
