title: A 27-mer oligonucleotide probe for the detection and measurement of the mRNA for uncoupling protein in brown adipose tissue of different species.
abstract: 1. Data from rats, cattle, mice, rabbits and humans indicate considerable species heterogeneity in the sequence of the gene coding for the mitochondrial uncoupling protein (UCP) in brown adipose tissue. A 27-base sequence of an exon region of the gene is, however, identical in rats and cattle; in mice, rabbits, and humans this same region shows only a single base difference from the sequence in rats and cattle. 2. A 27-mer oligonucleotide (3'-TGGAAGGGCGACCTGTGGCGGTTTCAG-5') complementary to the conserved region of the rat and cattle UCP genes has been synthesized as a potential probe for UCP mRNA in widely differing species. 3. Northern blots of RNA from rat brown fat showed that the oligonucleotide hybridized with a 1.5 kbase mRNA, indicative of UCP mRNA. No hybridization was observed with RNA from white fat (subcutaneous, internal), liver, kidney, skeletal muscle, heart and brain. Acute cold-exposure of rats and mice led to an increase in UCP mRNA level, while streptozotocin-induced diabetes resulted in a decrease. 4. The oligonucleotide hybridized with a 1.5 (or 1.9) kbase mRNA from brown fat of rats, mice, golden hamsters, Djungarian hamsters, and newborn rabbits, pipistrelle bats, lambs, goats and red deer. 5. The 27-mer oligonucleotide provides a simple probe for UCP mRNA across a wide range of mammals, obviating any need to obtain species-specific cDNAs.
authors.affiliation: Division of Biochemical Sciences, Rowett Research Institute, Bucksburn, Aberdeen, Scotland, U.K.
authors.forename: F
authors.initials: F
authors.lastname: Brander
authors.forename: J S
authors.initials: JS
authors.lastname: Keith
authors.forename: P
authors.initials: P
authors.lastname: Trayhurn
mesh_terms: D002001:Adipose Tissue, Brown / Q000737:chemistry*; D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics*; D002417:Cattle; D002685:Chiroptera; D006224:Cricetinae; D003670:Deer; D006041:Goats; D006801:Humans; D007473:Ion Channels; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000737:chemistry*; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits; D051381:Rats; D012756:Sheep; D013045:Species Specificity; D000071256:Uncoupling Protein 1
chemical_list: D002352:Carrier Proteins; D007473:Ion Channels; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D000071256:Uncoupling Protein 1