Application of polymerase chain reaction techniques to study of rabbit renin gene expression

Studies were performed to develop techniques to assess renin mRNA in a single microdissected juxtaglomerular apparatus (JGA) of rabbit using the polymerase chain reaction (PCR) method of amplification of cDNA sequences. In preliminary studies synthetic oligonucleotide primers corresponding to regions in the rat renin cDNA sequence, which are highly conserved between mouse, rat, and man, were found to yield good amplification efficiency with a rat renin cDNA template, but little product was observed with rabbit cDNA template. We therefore employed a nested primer PCR cloning technique to clone an 839 base pair portion of the rabbit renin cDNA to obtain species-specific sequence information for primer design. Here we report the nucleotide sequence of a partial rabbit renin cDNA clone and the use of species-specific primers that permit semiquantitative assessment of rabbit mRNA levels in the single JGA.