5 008 chow

All the experiments were carried out according to the Norma Oficial Mexicana (NOM-062-ZOO-1999, 9.4.2.1.3.) for the care and handling of laboratory animals. The protocols were reviewed and approved by the Animal Care and Use Committee of the Benemerita Universidad Autonoma de Puebla, identification code: BERRSAL71, 18-05-2017. Experiments were carried out in male ZDF rats (3 months old) from Charles River Laboratories, California, U.S.A. Throughout the text, diabetic-obese ZDF rats (ZDF-Lepr^fa/fa) will be designated as OZDF rats, and lean controls, non-obese non-diabetic ZDF (ZDF-Lepr^+/+) as LZDF. The rats were kept at the University Animal Core Facilities under controlled environmental temperature and, exposed to light-dark cycles of 12 h, with ad libitum consumption of water and high energy diet, Purina 5,008 chow.

All procedures involving animals were reviewed and approved by the Pfizer Institutional Animal Care and Use Committee. Lean and obese ZSF1 littermates were obtained from Charles River Laboratories at 8 weeks of age (strain codes #379 and #378, respectively). A total of 60 male rats, 30 lean and 30 obese, were enrolled in the study. Animals were housed and acclimated under 12 hour light-dark cycles with free access to water and Purina 5008 chow. 24 hour urine samples were collected from animals in metabolic cages prior to the termination of each cohort at 12, 20, 24, 29, 34, and 41 weeks of age. Between 34 and 41 weeks of age, one lean animal was euthanized for hematuria, and two obese animals were found dead of unknown cause with no outward clinical signs. Animals were sacrificed by CO₂ asphyxiation, and blood was collected via cardiac puncture. Kidney sections were immersion fixed for slide preparation, stored on ice for isolation of glomerular-enriched tissue, or snap frozen in liquid nitrogen for additional analyses.