Regulation of uncoupling proteins 2 and 3 in porcine adipose tissue.
This study was performed to determine whether or not uncoupling protein 2 (UCP2) and UCP3 expression in porcine subcutaneous adipose tissue are hormonally regulated in vitro and whether their expression is correlated with changes in metabolic activity. Tissue slices (approximately 100 mg) were placed in 12-well plates containing 1 mL of DMEM/F12 with 25 mM Hepes, 0.5% BSA, pH 7.4. Triplicate slices were incubated with basal medium or hormone supplemented media at 37 degrees C with 95% air/5% CO2. Parallel cultures were maintained for either 2 or 24 h to evaluate metabolic viability of the tissue. Slices were transferred to test tubes containing 1 mL of DMEM/F12 with 25 mM Hepes, 3% BSA, 5.5 mM glucose, 1 microCi 14C-U-glucose/mL and incubated for an additional 2 h at 37 degrees C. Glucose metabolism in 2-h incubations did not differ from 24-h (chronic) incubations, indicating viability was maintained (P>0.05). Expression of UCP2 and UCP3 was assessed in slices following 24h of incubation with various combinations of hormones by semi-quantitative RT-PCR. Expression of UCP2 was induced by leptin (100 ng/mL; P<0.05). Growth hormone (100 ng/mL) inhibited UCP2 expression (P<0.05). Expression of UCP3 was inhibited by growth hormone (100 ng/mL; P<0.05), tri-iodothyronine (10 nM; P<0.05) or leptin (100 ng/mL; P<0.05). Changes in UCP expression could not be associated with overall changes in glucose metabolism by adipose tissue slices in chronic culture.
Ramsay
T G
TG
Growth Biology Laboratory, USDA-ARS, BARC-East, Bldg. 200, Rm. 207, Beltsville, MD 20705, USA. tramsay@anri.barc.usda.gov
Rosebrough
R W
RW
D000273:Adipose Tissue / Q000378:metabolism*; D000818:Animals; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D003907:Dexamethasone / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology*; D005947:Glucose / Q000378:metabolism; D013006:Growth Hormone / Q000378:metabolism; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism; D007473:Ion Channels; D020738:Leptin / Q000378:metabolism; D008297:Male; D026901:Membrane Transport Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000737:chemistry / Q000235:genetics; D011897:Random Allocation; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D013552:Swine / Q000378:metabolism / Q000502:physiology*; D014284:Triiodothyronine / Q000378:metabolism; D000071246:Uncoupling Protein 2; D000071247:Uncoupling Protein 3
D002352:Carrier Proteins; D007328:Insulin; D007473:Ion Channels; D020738:Leptin; D026901:Membrane Transport Proteins; D024101:Mitochondrial Proteins; D000071246:Uncoupling Protein 2; D000071247:Uncoupling Protein 3; D014284:Triiodothyronine; D012313:RNA; D003907:Dexamethasone; D013006:Growth Hormone; D005947:Glucose