PPARγ Agonists Down-Regulate the Expression of Atp10c mRNA during Adipogenesis

Abstract

Abstract:We have shown that Atp10c, a type 4 P-type ATPase, is a strong candidate affecting glucose and lipid metabolism in humans and mice. Atp10c is a putative phospholipid translocase associated with cell signaling and intracellular protein trafficking. In order to examine the biological role of Atp10c, semiquantitative reverse transcriptasepolymerase chain reaction was carried out. Atp10c mRNA is expressed in 3T3-L1 cells and in primary preadipocytes and adipocytes generated from mice. Atp10c mRNA is regulated during fat cell differentiation and modulated by PPAR agonists and antagonists as well as by hormonal factors (insulin and dexamethasone). Atp10c expression is regulated by both the process of adipocyte differentiation and by effectors of fat and glucose metabolism. Taken together these data along with the published phenotype of the Atp10c heterozygote mice suggest that ATP10C is a newly identified protein with a possible biological role in the development of obesity and obesity-related metabolic disorders.

Transient Silencing of a Type IV P-Type ATPase,Atp10c, Results in Decreased Glucose Uptake in C2C12 Myotubes

“…Our laboratory has demonstrated that Atp10c heterozygous mice are insulin resistant and have an altered insulin-stimulated response in peripheral tissues. Our obesity mouse model is dietinduced and shows insulin resistance characterized by hyperinsulinemia, hyperglycemia, hyperlipidemia, and obesity in association with glucose intolerance [ 2 , 3 , 11 ]. In fact, a recent publication has sited ATP10C as a potential biomarker for obesity and related metabolic disorders [ 32 ].…”

“…The following procedures were performed as described elsewhere [ 5 , 10 , 11 ]. Cells were washed with ice cold 1X PBS, and total RNAs were isolated using RNeasy Mini RNA kit (Qiagen), according to the manufacturer's instructions.…”

“…Glucose uptake in myotubes was measured as previously described [ 11 ]. Briefly, myoblasts were plated in 6-well cell culture plates at a density of 2 × 10 5 cells/well and allowed to differentiate under normal conditions.…”

Equine peripheral blood‐derived mesenchymal stem cells: Isolation, identification, trilineage differentiation and effect of hyperbaric oxygen treatment

“…Stromal vascular fraction cells from adipose tissue were isolated as previously described with slight modifications [3,18–20]. Adipose tissue was rinsed with a 10% bleaching solution for disinfection and, after washing thoroughly with 1× phosphate‐buffered saline (PBS) a , minced into small pieces.…”

“…The medium was changed every 3 days. Adipocytes were stained with Oil Red O after 0, 2, 4 and 6 days, as described previously [20]. For staining, cells were fixed with 10% formalin for 15 min and rinsed with deionised water, followed by the addition of 2 aliquots of 85% propylene glycol h for 5 min each.…”