Murine and pediatric myocardial growth factor mRNA expression using reverse transcription-polymerase chain reaction

We utilized reverse transcription-polymerase chain reaction (RT-PCR) to examine the myocardial mRNA expression of growth factors in mice and children. Total cellular RNA was extracted from these tissues using acidified-phenol guanidinium thiocyanate. Optimal oligonucleotide primer pairs for RT-PCR were selected with the aid of the computer program Oligo 4.0. RT-PCR analysis of total cellular RNA demonstrated the measurable presence of the mRNA for the following growth factors in the myocardium of both mice and children: acidic fibroblast growth factor (aFGF), basic FGF (bFGF), insulin-like growth factor (IGF)-1, IGF-2, platelet-derived growth factor (PDGF)-A chain, PDGF-B chain, transforming growth factor (TGF)beta-1, TGFbeta-2, and TGFbeta-3. The amplified cDNA message for the growth factors and beta-actin migrated as a discrete band at the expected base pair number on agarose gels stained with the intercalating fluorescent dye ethidium bromide. Densitometry of the photographic negatives of these gels permitted the rapid and semiquantitative comparison of these factors. These data demonstrate the feasibility and reproducibility of utilizing RT-PCR for the specific detection and semiquantitation of mRNA expression of myocardial aFGF, bFGF, PDGF-A chain, PDGF-B chain, lGF-1, lGF-2, TGFbeta-1, TGFbeta-2, and TGFbeta-3 in mice and children.