抄録
Exposure to cold potentiates cell proliferation and differentiation in brown adipose tissue (BAT) by activating the sympathetic nerve systems, and causes an increase in mitochondrial biogenesis as well as the activation of genes encoding several proteins. We investigated the expression of proteins on Xenopus laevis oocytes injected with total RNA purified from the cold acclimated BAT by immuno electron microscopy, and further the induction in UCPs (UCP1, 2, 3) in the BAT after cold acclimation of rats by Real-Time PCR technique.
Total RNA from rat interscapular BAT of warm- or cold-acclimated rats was extracted using the AGPC method. RNA was digested with RNase-free DNase to remove eventual traces of genomic DNA. mRNA was isolated by poly A selection and then double stranded cDNA was synthesized. Immuno-gold electron microscopy using anti UCP polyclonal antibody revealed the localization of immuno-gold particles on the inner membrane of mitochondria like structure of the newly synthesized membrane in the oocytes injected with the cold acclimated rat transcript. Real-Time PCR assay that combines TaqMan technology with fluorescent-based high resolution capillary electrophoresis and GeneScan DNA fragment analysis was applied for detection and quantification of BAT UCP mRNA. Real-Time assays were performed using primers corresponding to conserved domains of rat UCP1, UCP2 and UCP3. The result revealed that UCP2 transcripts were elevated in the RNA fraction purified from BAT of cold acclimated rats than that of warm acclimated rats. [Jpn J Physiol 54 Suppl:S245 (2004)]
