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Differential gene expression following early renal ischemia/reperfusion
Suroj Supavekin1, Weijia Zhang, Raju Kucherlapati
1Pediatric Nephrology, Children's Hospital at Montefiore, Albert Einstein College of Medicine, Bronx, New York, USA.
Kidney International
|April 5, 2003
Summary
Early renal ischemia/reperfusion injury triggers tubule cell apoptosis via activated death receptor and mitochondrial pathways. Gene expression analysis reveals key proapoptotic factors, highlighting mechanisms for cell loss and potential repair.
Area of Science:
- Nephrology
- Molecular Biology
- Cellular Biology
Background:
- Acute renal failure (ARF) following ischemia/reperfusion (I/R) injury involves tubule cell apoptosis.
- The precise molecular mechanisms driving this early cell death are not fully understood.
Purpose of the Study:
- To identify apoptosis-related genes with differential expression in the early stages of renal I/R injury.
- To elucidate the molecular pathways involved in early tubule cell apoptosis post-ischemia.
Main Methods:
- Mice underwent unilateral renal artery clamping followed by reperfusion.
- Tubule cell apoptosis was assessed using DNA laddering and TUNEL assays.
- Global gene expression changes were analyzed using cDNA microarrays, with validation by RT-PCR and immunohistochemistry.
Main Results:
- Microarray analysis identified altered expression of transcription factors, growth factors, signal transduction molecules, and apoptotic factors.
- Key proapoptotic genes, including FADD, DAXX, BAD, BAK, and p53, were significantly upregulated.
- Semiquantitative RT-PCR and immunohistochemistry confirmed the upregulation of these proapoptotic genes.
Conclusions:
- Apoptosis is a critical mechanism contributing to early tubule cell loss after renal I/R injury.
- Both death receptor-dependent (FADD-DAXX) and mitochondrial (BAD-BAK) apoptotic pathways are activated.
- Upregulation of genes involved in growth, proliferation, transcription, and cytoskeletal factors suggests inherent mechanisms for renal tubule cell repair and regeneration.