An anti N-terminal antibody localizes NBCn1 to rat renal and cardiac blood vessels

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    The electroneutral sodium bicarbonate cotransporter NBCn1 protein was localized in basolateral membrane of medullary thick ascending limbs, intercalated cells and later in duodenum and choroids plexus. NBCn1 was originally cloned from rat aorta and mRNA was detected in e.g. heart, spleen and aorta. However, the protein has not been detected in these tissues. Therefore, a 20 amino acid antibody was designed against n-terminal of rat NBCn1. The antibody was affinity purified and validated by immunoblotting, recognizing an approximately 150 kDa protein in rat kidney, cerebrum, cerebellum and duodenum. Preabsorption of the antibody with the immunizing peptide prevented detection of NBCn1. Interestingly we also found an approximately 150kDa band in aorta, both ventricles and atrium of the heart, mesenteric arteries, lung, spleen, liver, pylorus, jejunum, ileum, colon, pancreas, cerebrum, cerebellum, epididymis, parotid gland and inner medulla(IM) and cortex of the kidney. Immunohistochemical analyzis confirmed the previously described labeling in medullary thick ascending limb of the kidney, duodenum and the choroids plexus. We also found labeling of renal small arteries, and capillaries of the heart ventricles. Thus, the development of the n-terminal antibody allowed the localization of NBCn1 protein to major cardiovascular tissues. In conclusion, the findings agree, and possibly explains, the reported electroneutral Na+,HCO3- cotransport in vascular smooth muscle and heart.

     

    Periode5 apr. 2005
    Begivenhedstitel Experimental Biology 2005
    BegivenhedstypeKonference
    ArrangørFASEB (American Physiological Society) - IUPS joint meeting
    PlaceringSan Diego, USAVis på kort