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Updated: Jun 27, 2026
5/6th Nephrectomy in Combination with High Salt Diet and Nitric Oxide Synthase Inhibition to Induce Chronic Kidney Disease in the Lewis Rat
Published on: July 3, 2013
Splice variants of neuronal nitric oxide synthase are present in the rat kidney
Cheryl Smith1, Michael Merchant, Andrea Fekete
1Department of Physiology and Functional Genomics, University of Florida, POB 100274, Gainesville, FL 36210, USA.
Background:
Decreased renal cortical neuronal NO synthase (nNOS) abundance/activity correlates with progression of chronic kidney disease (CKD) in a number of animal models.
Methods:
Western blotting with both N-terminal and C-terminal antibodies, immunoprecipitation, proteomics, RT-PCR and in situ hybridization were used to identify nNOS splice variants in the rat kidney.
Results:
We have identified two nNOS proteins and transcripts in the rat kidney; nNOSalpha (approximately 160 kDa) and nNOSbeta (approximately 140 kDa), a catalytically active exon-2 deletion variant, lacking both the PDZ and protein inhibitor of nNOS (PIN) domains. We also report that nNOSbeta protein abundance is increased in the kidney at 11 weeks following 5/6th nephrectomy (5/6NX)-induced CKD while nNOSalpha protein abundance is diminished. The transcript data parallel the protein data in 5/6NX. By in situ hybridization, there is abundant nNOSalpha mRNA widely distributed throughout the normal kidney cortex, with very sparse nNOSbeta mRNA confined to a few proximal tubules. In a second injury model (6 weeks after 5/6 renal mass reduction by combined right kidney ablation and infarction of approximately 2/3 of the left kidney; 5/6 A/I), nNOSalpha mRNA almost disappears from the kidney cortex while nNOSbeta mRNA abundance increases in tubules and tubulo-interstitium.
Conclusion:
The renal cortical nNOSbeta protein is present in low abundance in the normal kidney and increases with injury, in an inverse pattern of change with the nNOSalpha.
