Methods for assessing and modulating UCP2 expression and function.
Uncoupling protein 2 (UCP2) is a member of the uncoupling protein family. It is expressed in the inner mitochondrial membrane and plays a role in the control of free radical production, oxidative damage, insulin secretion, and fatty-acid peroxide exportation. Although UCP2 expression occurs in several tissues, some of its most remarkable functions are exerted in organs of difficult experimental access, such as the central nervous system, particularly the hypothalamus and the pancreatic islets. In addition, due to its low levels of expression in the mitochondrial membrane, studying UCP2 expression and function depends on specific- and well-established methods. This chapter describes methods for directly assessing UCP2 expression and function in different tissues. Purified mitochondria preparations are used for enhancing the capacity of detection of UCP2 protein or for evaluating the role of UCP2 in mitochondria respiration. Exposure of experimental animals to cold environment leads to increased UCP2 expression, while reduction of its expression can be achieved directly by targeting its mRNA with antisense oligonucleotides, or indirectly by targeting PGC-1alpha expression with antisense oligonucleotides.
Velloso
Lício A
LA
Department of Internal Medicine, University of Campinas, Campinas, SP, Brazil.
Degasperi
Giovanna R
GR
Vercesi
Aníbal E
AE
Saad
Mário A
MA
D000818:Animals; D016375:Antisense Elements (Genetics); D003080:Cold Temperature; D005786:Gene Expression Regulation; D007031:Hypothalamus / Q000378:metabolism; D015151:Immunoblotting; D007473:Ion Channels / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D007515:Islets of Langerhans / Q000378:metabolism; D008297:Male; D051379:Mice; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism; D000071248:Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; D016133:Polymerase Chain Reaction; D016601:RNA-Binding Proteins / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D014157:Transcription Factors / Q000235:genetics; D000071246:Uncoupling Protein 2
D016375:Antisense Elements (Genetics); D007473:Ion Channels; D024101:Mitochondrial Proteins; D000071248:Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; C501429:Ppargc1a protein, rat; D016601:RNA-Binding Proteins; D014157:Transcription Factors; C000604390:Ucp2 protein, mouse; C000604391:Ucp2 protein, rat; D000071246:Uncoupling Protein 2; D010100:Oxygen