A semi-quantitative method for the estimation of adenosine A1 receptor mRNA levels in rat kidney

OBJECTIVE: To develop Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) method for the estimation of adenosine A₁ receptor mRNA levels in rat kidney. MATERIAL AND METHODS: Total cellular RNA was isolated from whole rat kidney by small-scale total RNA preparation protocol and it was reverse transcribed into cDNA. The cDNA was subjected to PCR amplification using gen specific primers. The amplified cDNA was evaluated by gel electrophoresis and the intensity of the bands were visualized and quantitated with a FujiBAS 1000 PhosphorImager. Then the adenosine A₁ receptor mRNA levels were extrapolated from the standard curve. RESULTS: Adenosine A₁ receptor mRNA levels in rat kidney were measured as: 1.30 ± 0.17 X 10⁷ copies of adenosine A₁ receptor transcript/mg total RNA. CONCLUSION: The RT-PCR method developed for the estimation of adenosine A₁ receptor mRNA levels in rat kidney is sensitive and reliable.