Abstract
Liver homogenates produced from fasted and non-fasted C57BL/6J female mice were assayed for total lipolytic activity measured as hydrolysis of [9,10-3H(N)]-triolein into [3H] free fatty acids (FFA). Liver homogenates were also used for immunoblotting to determine levels of the lipolytic enzymes adipose-triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL), as well as site specific phosphorylation at the 14-3-3 binding site of ATGL and the serine 565 and serine 660 sites of HSL. Significantly higher triolein hydrolysis activity was observed in fasted liver samples, as well as a significant increase in total ATGL and a significant decrease in HSL phosphorylation at the S565 site.
Highlights
University of Waterloo, Department of Kinesiology, Faculty of Applied Health Sciences, 200 University Avenue W., BMH1110, Waterloo, Ontario, Canada N2L 3G1 article info
Liver homogenates produced from fasted and non-fasted C57BL/6J female mice were assayed for total lipolytic activity measured as hydrolysis of [9,10-3H(N)]-triolein into [3H] free fatty acids (FFA)
Higher triolein hydrolysis activity was observed in fasted liver samples, as well as a significant increase in total adipose-triglyceride lipase (ATGL) and a significant decrease in hormone-sensitive lipase (HSL) phosphorylation at the S565 site
Summary
C57BL/6J mice were bred in the central animal facilities at the University of Waterloo from founders originally purchased from Jackson Laboratories (Bar Harbor, Maine, USA). Fourteen to sixteen week old female mice were group housed in a temperature and humidity controlled environment with a standard 12 h light: h dark cycle. Standardized rodent diet (Research Diets #D12450H, New Brunswick, New Jersey, USA) and water were provided to the mice ad libitum for 9 days. Half of the mice were randomly chosen for a 16 h overnight fast from 5 pm to 9 am prior to cervical dislocation and tissue collection as we have previously described [1]. Protocols were approved by the Animal Care Committee at the University of Waterloo, with all procedures performed in accordance with the Canadian Council on Animal Care
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