RQ-PCR analysis of L-type calcium channel transcripts in ON, and corresponding ON-protein expression. A. RQ-PCR analysis. Gene-specific primers for the α1A and α1D subunits of the L-type calcium subunits were generated from Genbank. RQ-PCR results were internally normalized using primers for cyclophilin. There is 7-10 fold less α1A and 5-7 fold less α1D mRNA in human and rhesus macaque ON than in rat ON. B. Western analysis. ON homogenates from rat brain (RBr), rat ON (RON), monkey ON (MON) and human ON (HON) were subjected to PAGE, transferred to PVDF membrane, and probed with a mouse monoclonal antibody to the α1D subunit of the L-type calcium channel. The specific protein (170kD; arrow) is detectable as a strong band present in the rat brain and -ON homogenates. Signal strength for the α1D subunit is considerably less in homogenates from monkey and human ON. The lower inset band is relative β-actin signal from each lane.