Detection of brown adipose tissue uncoupling protein mRNA in adult patients by a human genomic probe.
1. Studies on human brown adipose tissue require specific molecular probes. A human genomic library has been screened with a complementary DNA corresponding to the uncoupling protein (UCP) of rat brown adipose tissue mitochondria. 2. Two recombinant phages were isolated, carrying genomic sequences of human UCP. From them we have subcloned a 0.5 kilobase fragment. This fragment, H-Ucp-0.5, contained two intronic regions and two exonic regions. Exonic regions encoded a sequence of 84 amino acids which exhibited a strong homology with central domain at rat UCP. The organization of H-Ucp-0.5 was confirmed by SI mapping analysis. 3. A Southern analysis suggested that the gene is single type in the human, as it is in rodents. 4. In Northern analysis experiments, H-Ucp-0.5 detected a specific 1.8 kb mRNA in human brown adipose tissue obtained from six patients with phaeochromocytoma and from one patient with a hibernoma. This molecular probe is a new, sensitive and reliable tool with which to study human brown adipocytes.
Bouillaud
F
F
Centre de Recherche sur la Nutrition, Centre National de la Recherche Scientifique, Meudon, France.
Villarroya
F
F
Hentz
E
E
Raimbault
S
S
Cassard
A M
AM
Ricquier
D
D
D002001:Adipose Tissue, Brown / Q000032:analysis*; D000328:Adult; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins; D004247:DNA; D005819:Genetic Markers; D006801:Humans; D007473:Ion Channels; D008565:Membrane Proteins / Q000235:genetics*; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D051381:Rats; D000071256:Uncoupling Protein 1
D002352:Carrier Proteins; D005819:Genetic Markers; D007473:Ion Channels; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D012333:RNA, Messenger; D000071256:Uncoupling Protein 1; D004247:DNA