All protocols followed Canadian Council on Animal Care guidelines and were approved by the University of Guelph and York University Animal Care Committees. For co-culture experiments, five 10 week old ZDF and five age matched lean Zucker rats (Charles River, St. Constant, QC, Canada) were singly housed in standard clear, plastic cages. Male rats were used to remove any effects of estrogen on the adipose tissue adipokine secretion profile as previously described [15 (link)]. Animals habituated in a temperature and humidity (50–60%) controlled room for seven days and were fed a standard show diet (Purina 5012 Chows, Ralston Purina, St. Louis, MO) and water ad libitum.
For RSV supplementation experiments, four-week old male ZDF rats were housed as previously described [21 (link)]. Following acclimatization, animals were fed either a standard powdered chow diet (Purina 5008 diet; Purina; ZDF) or a diet supplemented with ~200 mg/kg body weight RSV (Cayman Chemical, Ann Arbor, MI, USA; ZDF+RSV) for 6 weeks (4). While this dose of RSV is likely not attainable through diet alone, it is similar to many other rodent-based reports in the literature [20 (link),22 (link),23 (link)]. Zucker rats served as lean controls.
For RSV supplementation experiments, four-week old male ZDF rats were housed as previously described [21 (link)]. Following acclimatization, animals were fed either a standard powdered chow diet (Purina 5008 diet; Purina; ZDF) or a diet supplemented with ~200 mg/kg body weight RSV (Cayman Chemical, Ann Arbor, MI, USA; ZDF+RSV) for 6 weeks (4). While this dose of RSV is likely not attainable through diet alone, it is similar to many other rodent-based reports in the literature [20 (link),22 (link),23 (link)]. Zucker rats served as lean controls.