Figure 3. Gene expression of A₃ adenosine receptor in captured cells from the ganglion cell layer

Expression of A₃ adenosine receptor mRNA was detected from material obtained from labeled cells in the ganglion cell layer. Panels A-C shows results from traditional PCR run on 1% agarose gels. A: The 641 bp PCR product of the 5' end of the encoding sequence from primer pair A₃U is shown. B: The 724 bp PCR product of the 3' portion from primer pair A₃L. C: The 123 bp PCR product of the cDNA of A₃ adenosine receptor using the primers for real-time PCR. The left lanes correspond to PCR marker; size of the DNA fragments is indicated on the left. Negative controls, obtained by omitting the reverse transcriptase, are indicated by "RT-". D: A typical result from real-time PCR analysis using primers for the GAPDH and A₃ receptor genes. The abscissa indicates the number of PCR cycles and the ordinate shows the fluorescence emitted by SYBR green dye-labeled PCR products. The curves on the left represent the PCR product of GAPDH and the curves on the right represent the PCR products of A₃ adenosine receptor. Each reaction has been performed here in triplicate. Similar results were found with RNA from 5 different microdissections.