Visualization of Intracellular ATP Dynamics in Different Nephron Segments Under Pathophysiological Conditions Using the Kidney Slice Culture System

Authors

• Shigenori Yamamoto
• Shinya Yamamoto
• Masahiro Takahashi
• Akiko Mii
• Akihiro Okubo
• Naoya Toriu
• Shunsaku Nakagawa
• Takaaki Abe
• Shingo Fukuma
• Hiromi Imamura
• Masamichi Yamamoto
• Motoko Yanagita

Language

English

Publication Date

9-1-2024

Journal

Kidney International

DOI

10.1016/j.kint.2024.05.028

PMID

38996810

Abstract

ATP depletion plays a central role in the pathogenesis of kidney diseases. Recently, we reported spatiotemporal intracellular ATP dynamics during ischemia reperfusion (IR) using GO-ATeam2 mice systemically expressing an ATP biosensor. However, observation from the kidney surface did not allow visualization of deeper nephrons or accurate evaluation of ATP synthesis pathways. Here, we established a novel ATP imaging system using slice culture of GO-ATeam2 mouse kidneys, evaluated the ATP synthesis pathway, and analyzed intracellular ATP dynamics using an ex vivo IR-mimicking model and a cisplatin nephropathy model. Proximal tubules (PTs) were found to be strongly dependent on oxidative phosphorylation (OXPHOS) using the inhibitor oligomycin A, whereas podocytes relied on both OXPHOS and glycolysis using phloretin an active transport inhibitor of glucose. We also confirmed that an ex vivo IR-mimicking model could recapitulate ATP dynamics in vivo; ATP recovery in PTs after reoxygenation varied depending on anoxic time length, whereas ATP in distal tubules (DTs) recovered well even after long-term anoxia. After cisplatin administration, ATP levels in PTs decreased first, followed by a decrease in DTs. An organic cation transporter 2 inhibitor, cimetidine, suppressed cisplatin uptake in kidney slices, leading to better ATP recovery in PTs, but not in DTs. Finally, we confirmed that a mitochondria protection reagent (Mitochonic Acid 5) delayed the cisplatin-induced ATP decrease in PTs. Thus, our novel system may provide new insights into the energy dynamics and pathogenesis of kidney disease.

Keywords

Animals, Adenosine Triphosphate, Cisplatin, Oxidative Phosphorylation, Kidney Tubules, Proximal, Mice, Glycolysis, Podocytes, Reperfusion Injury, Disease Models, Animal, Cimetidine, Male, Kidney Tubules, Distal, Organ Culture Techniques, Mice, Transgenic, Oligomycins, Phloretin, Mice, Inbred C57BL, AKI, ATP, cisplatin nephropathy, energy dynamics, imaging, mitochondria

Published Open-Access

yes

Recommended Citation

Yamamoto, Shigenori; Yamamoto, Shinya; Takahashi, Masahiro; et al., "Visualization of Intracellular ATP Dynamics in Different Nephron Segments Under Pathophysiological Conditions Using the Kidney Slice Culture System" (2024). Faculty, Staff and Students Publications. 8713.
https://digitalcommons.library.tmc.edu/baylor_docs/8713