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Kidney Week

Abstract: FR-OR111

Whole-Kidney Three-Dimensional Imaging Reveals the Progression of Renal Sympathetic Denervation after Ischemia-Reperfusion Injury

Session Information

Category: CKD (Non-Dialysis)

  • 1903 CKD (Non-Dialysis): Mechanisms

Authors

  • Hasegawa, Sho, The University of Tokyo Graduate School of Medicine, Tokyo, Japan
  • Tanaka, Tetsuhiro, The University of Tokyo Graduate School of Medicine, Tokyo, Japan
  • Susaki, Etsuo A., The University of Tokyo Graduate School of Medicine, Tokyo, Japan
  • Komaba, Hirotaka, Tokai University School of Medicine, Isehara, Japan
  • Wada, Takehiko, Tokai University School of Medicine, Isehara, Japan
  • Saito, Hisako, The University of Tokyo Graduate School of Medicine, Tokyo, Japan
  • Sugahara, Mai, The University of Tokyo Graduate School of Medicine, Tokyo, Japan
  • Uchida, Lisa, The University of Tokyo Graduate School of Medicine, Tokyo, Japan
  • Ito, Marie, The University of Tokyo Graduate School of Medicine, Tokyo, Japan
  • Fukagawa, Masafumi, Tokai University School of Medicine, Isehara, Japan
  • Ueda, Hiroki R., The University of Tokyo Graduate School of Medicine, Tokyo, Japan
  • Nangaku, Masaomi, The University of Tokyo Graduate School of Medicine, Tokyo, Japan
Background

The sympathetic nervous system is critical in maintaining the homeostasis of renal functions. Its three-dimensional (3D) structures in the kidney, however, have not been elucidated because of the limitation of conventional imaging methods. CUBIC (Clear, Unobstructed Brain/Body Imaging Cocktails and Computational analysis) is a newly developed tissue clearing technique, which enables 3D imaging of whole-organ with single-cell resolution. Here, we apply CUBIC to the kidney research, visualizing the 3D structure of renal sympathetic nerves, showing the process of denervation after ischemia-reperfusion injury (IRI).

Methods

Eight-week-old male C57BL/6 mice were subjected to sham operation or bilateral IRI for 45 minutes, and sacrificed 28 days after surgery. The kidneys were optically cleared using CUBIC protocol. After making kidneys transparent, the sympathetic nerves were labeled with 3D immunofluorescent staining. Whole-kidney imaging was acquired by a light-sheet fluorescent microscopy.

Results

The spatial distribution of sympathetic nerves was clearly visualized. Compared with the sham group (figure 1A), the nerve density in the cortical lesion was significantly decreased in the IRI group (figure 1B).

Conclusion

The new 3D imaging technique has demonstrated the sustained renal sympathetic denervation after IRI. Dysfunction of sympathetic nerves might be related to the progression of chronic kidney disease.

Figure 1. The representative 3D imaging of sympathetic nerves in the kidney. (A) Sham-operated group, (B) Bilateral IRI group.

Funding

  • Government Support - Non-U.S.