Abstract
The aim of this study was to determine the expression of the endothelin receptor subtype mRNAs in human detrusor cultured smooth muscle cells using reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization (ISH). First strand cDNA was made from human detrusor cultured smooth muscle cells total RNA and used for PCR with primers designed to amplify fragments of the ET<sub>A</sub> and ET<sub>B</sub> endothelin receptor subtype cDNA sequences. Subcloned fragments of the ET<sub>A</sub> and ET<sub>B</sub> endothelin receptor cDNAs were used to synthesize digoxigenin-labeled cRNA probes by in vitro transcription. COS-7 cells transfected with the ET<sub>A</sub> and ET<sub>B</sub> receptor cDNAs were used as positive control and to confirm the absence of cross-hybridization due to sequence homology. Both ET<sub>A</sub> and ET<sub>B</sub> receptor mRNAs were detected by RT-PCR analysis. By ISH, both ET<sub>A</sub> and ET<sub>B</sub> receptor subtype mRNAs were detected. However, ET<sub>A</sub> signal was much more intense than ET<sub>B</sub> signal. These results indicate that mRNAs for both ET<sub>A</sub> and ET<sub>B</sub> receptors are expressed in detrusor smooth muscle cells of human urinary bladder. The ET<sub>A</sub> receptor is the predominant detrusor ET receptor.
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