Adiponectin, an insulin-sensitizing factor secreted from adipose tissue, is decreased in individuals with type 2 diabetes (T2D) and increased in response to thiazolidinedione (TZD) therapy. Changes in its secretion and assembly into higher-order forms affect insulin sensitivity. To determine the relative potency of TZDs on intra-adipocyte multimerization and secretion of adiponectin, we assessed the impact of in vivo low-or high-dose rosiglitazone treatment alone or combined with metformin in subjects with T2D. T2D subjects received high-dose rosiglitazone (8 mg/day), high-dose metformin (2,000 mg/day), or low-dose combination rosiglitazone-metformin therapy (4 mg ϩ 1,000 mg/day) for 4 mo. All subjects were then switched to high-dose rosiglitazonemetformin combination therapy (8 mg ϩ 2,000 mg/day) for another 4 mo. Low-dose rosiglitazone increased serum adiponectin, whereas the high dose increased both adipocyte content and serum adiponectin levels. TZDs selectively increased the percentage of circulating adiponectin in the potent, high-molecular-weight (HMW) form. No TZD effects were evident on multimer distribution in the cell. Expression of the chaperone protein ERp44, which retains adiponectin within the cell, was decreased by TZD treatment. No changes occurred in Ero1-L␣ expression. Metformin had no effect on any of these measures. Increases in adiponectin correlated with improvements in insulin sensitivity. In vivo, TZDs have apparent dose-dependent effects on cellular and secreted adiponectin. TZD-mediated improvements in whole body insulin sensitivity are associated with increases in circulating but not cellular levels of the HMW adiponectin multimer. Finally, TZDs promote the selective secretion of HMW adiponectin, potentially, in part, through decreasing the expression of the adiponectin-retaining protein ERp44. thiazolidinedione; adipocyte; ERp44; Ero1-L␣; chaperone proteins AS THE PREVALENCE OF OBESITY CONTINUES to rise at an alarming rate, so do associated rates of obesity-related metabolic dysfunction; among these are coagulation abnormalities, dyslipidemia, hyperinsulinemia, glucose intolerance, and type 2 diabetes (13, 34). Although long recognized to play a role in fat metabolism, the endocrine role of adipose tissue is a relatively recent and important discovery, potentially providing a link between obesity and disorders of metabolism. Among the growing list of identified adipose endocrine products, adiponectin is uniquely related to changes in insulin sensitivity. Low levels are seen in obesity and insulin-resistant states and are predictive of type 2 diabetes (15), whereas high levels are associated with leanness and greater insulin sensitivity (28, 40) and reduced risk for development of type 2 diabetes (14).Although synthesized as a 32-kDa monomeric protein, adiponectin undergoes extensive posttranslational processing and assembly into low-molecular-weight (LMW) trimers, middlemolecular-weight (MMW) heximers, and high-molecularweight (HMW) complexes prior to secretion (38). In vitro, the HM...