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Abstract: SA-OR019

Renal Ischemia Resistant Rat Strains and Ischemic Preconditioned (IPC) Kidneys Share Upregulated Mitochondria Proteins

Session Information

Category: Acute Kidney Injury

  • 103 AKI: Mechanisms

Authors

  • Kolb, Alexander Louis, Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Collett, Jason Andrieu, Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Witzmann, Frank, Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Aryal, Uma, Bindley Bioscience Center, Purdue University, West Lafayette, Indiana, United States
  • Basile, David P., Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Bacallao, Robert L., Indiana University School of Medicine, Indianapolis, Indiana, United States
Background


Brown-Norway (BN) rats are resistant to renal ischemia reperfusion injury as compared to Sprague-Dawley (SD) or Dahl Salt Sensitive (DS) rats. This resistance is linked to 3 chromosomes in BN rats. Also IPC kidney mitochondria have significant changes in their proteome. The study goal was to identify mitochondria proteins that are over-expressed in both IPC and genetically resistant tissues. Quantitative proteomic analysis of mitochondria was performed in IPC kidneys and compared to sham conditioned kidneys. A second set of comparisons was performed on kidney cortex mitochondria isolated from BN rats and DS rats.

Methods


Mitochondria were isolated from renal cortex of BN rats, DS rats, IPC and IPC sham treated kidneys of SD rats. 4 animals were used for each group with 3 replicant samples. Mitochondria proteomes were identifed using label free quantitative proteomic analysis using LC-MS/MS. LS-MS/MS data were collected in Thermo Q Exactive Orbitrap High Field (HF) MS coupled to Dionex UltiMate 3000 RP HPLC system. Raw data were analyzed using MaxQuant and Andromeda search engines. Highest stringency conditions were used in protein identification in all searches. Proteins were identified by searching against the rat protein sequences database from Uniprot. Relative protein quantitation was performed using the MaxQuant LQF algorithm.

Results


Greater than 1300 proteins were identified in the sham versus IPC mitochondria, 571 proteins are significantly overexpressed in the IPC group (p values <0.05). In the proteomic comparison between BN and DS rats, 1184 proteins were identified with high confidence. Of the 1184 proteins 770 proteins were expressed 1.8-fold or greater in BN versus SS rats (p values <0.05 ). Comparing the two groups revealed 78 common proteins. Ingenuity pathway analysis revealed enhanced pathways that include response to drug, negative regulation of apoptosis, protein transport, metabolic processes and redox homeostasis.

Conclusion


78 endogenous mitochondria proteins are significantly overexpressed in mitochondria from kidney cells in an ischemic preconditioned state or isolated from a rat strain with genetic resistance to ischemia. These proteins represent important targets for pharmacologic exploration to develop preventative agents for acute kidney injury.

Funding

  • Veterans Affairs Support