Measurement of gut hormone gene expression: mRNA and peptides.
During the past decade numerous methods for measurement of mRNA and peptides have been developed. Since the expression cascade from DNA to protein is regulated at all levels, the methods should be carefully designed to accomplish the purpose of the analysis. Regulation of the nuclear processing, the translational activity and the decay of a particular mRNA changes the proportionality between transcriptional activity and production of prepropeptide. Moreover, the post-translational maturation of the pro-hormones may be attenuated. Detection of mRNA is valuable and feasible because it is easy to generate cDNA probes for most hormones, and because mRNA demonstration unequivocally indicates the cellular site of gene expression. The deduction of preprohormone structures has also made it possible to improve the versatility of radioimmunoassays (RIA). Monospecific antibodies and pure tracers have allowed the development of sequence-specific RIA libraries for bioactive peptides and their precursors. Recently we have introduced a simple processing-independent analysis (PIA) for clinical use, since the post-translational maturation of gut peptides may be changed in gastrointestinal diseases. So far PIA has improved the diagnostic sensitivity for gut hormone tumours.
Nielsen
F C
FC
Department of Clinical Biochemistry, University of Copenhagen, Rigshospitalet, Denmark.
Rehfeld
J F
JF
D000818:Animals; D001681:Biological Assay; D015152:Blotting, Northern; D005768:Gastrointestinal Hormones / Q000096:biosynthesis* / Q000737:chemistry; D005786:Gene Expression Regulation; D006801:Humans; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000737:chemistry*; D011863:Radioimmunoassay; D014158:Transcription, Genetic
D005768:Gastrointestinal Hormones; D012333:RNA, Messenger