Abstract
Abstract Basic fibroblast growth factor (bFGF) is a multipotential peptide with trophic, mitogenic, and differentiating effects on the central and the peripheral nervous system. Expression of the mRNA for bFGF has been shown to be temporally and regionally regulated in developing and adult rat brain. Although there are discernible levels of bFGF protein in the late embryonic rat brain and high levels in the mature rat brain, bFGF mRNA levels are low and difficult to detect at all stages. A sensitive Northern blotting technique for imaging bFGF transcripts is described. Thin surface-tension gels allow maximum resolution of mRNAs. Multiple bFGF mRNA transcripts are detected in adult rat brain at 6.0, 3.7, 2.5, 1.8, 1.6, 1.4, and 1.0 kb in about equal amounts, while in embryonic rat brain, the 1.8-kb mRNA is the most abundant. Homology between bFGF and the 28S ribosomal RNA leads to a hybridization pattern that is difficult to interpret in the developing brain. The design of probes to avoid this nonspecific interaction is discussed. Methods for resolving sense and antisense mRNAs are described. The high-affinity bFGF receptor FGFR1, which has tyrosine-specific protein kinase activity, is expressed in both developing and adult rat brain, with a message size of 4.3 kb detected by Northern analysis. Finally, methods for measuring bFGF protein to correlate with the presence of mRNA in the developing brain are discussed.
Published Version
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call