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In vivo expression profile of a H⁺-K⁺-ATPase ₂-subunit promoter-reporter transgene

¹Internal Medicine and ²Integrative Biology and Pharmacology, and ³The Institute of Molecular Medicine for the Prevention of Human Diseases, The University of Texas Health Sciences Center at Houston, Houston, Texas 77030

Submitted 26 January 2003 ; accepted in final form 3 February 2004

Because little is known about the molecular basis of transcriptional regulation of the murine H⁺-K⁺-ATPase ₂ (HK₂) gene or other genes whose expression is restricted in part to the collecting duct, especially in vivo, we developed transgenic mice carrying an insertional HK₂ promoter-reporter gene construct. In these mice, the region –7,264/+253 of the HK₂ 5'-flanking region controls expression of the reporter gene enhanced green fluorescent protein (EGFP). Patterns of HK₂/EGFP transgene expression were examined by fluorescence microscopy and immunoblotting. Of 10 major organs examined, EGFP immunoreactivity was detected abundantly in the kidney, and to a far lesser extent, in the brain and lung. Within the kidney, EGFP fluorescence was detected exclusively in the collecting ducts of transgenic mice and colocalized with the cellular distribution of both endogenous HK₂ and aquaporin-2, consistent with the known expression pattern of endogenous HK₂ in principal cells. Surprisingly, no transgene expression was evident by immunoblotting or fluorescence microscopy in the distal colon, the site of the highest endogenous HK₂ expression. Although previous studies of steady-state mRNA levels suggested differences in HK₂ gene regulation in the kidney and colon, our results provide the first direct evidence of differential transcriptional control of the HK₂ gene in these organs and suggest that regions outside the 5'-flanking region or other regulatory factors play a role in HK₂ expression in the distal colon.

transgenic mice; gene regulation

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