5p76 prolab isopro rmh 3000

Mice were randomly assigned to receive the following treatments of n-3 PUFAs after TBI: (1) vehicle control: mice were fed a regular laboratory rodent diet (Prolab Isopro RMH 3000 5P76; LabDiet, St. Louis, MO, USA), in which the n-3 PUFA content was low (0.36%), and received intraperitoneal (IP) injections of 0.9% NaCl (300 µl per injection, 2 h after TBI and then daily for 14 days); (2) injections of mixed n-3 PUFAs (“N3”): mice were fed a regular diet and received IP injections of EPA and DHA (7 mg of EPA and 3 mg of DHA/kg body weight, diluted in 300 μl of 0.9% NaCl per injection, 2 h after TBI and then daily for 14 days); (3) fish oil dietary supplementation (FO): mice were fed a diet supplemented with n-3 PUFAs (DHA and EPA, triple-strength n-3 fish oil; Puritan’s Pride, Oakdale, NY, USA) to reach a 4% final n-3 PUFA concentration beginning 1 day after TBI and for up to 35 days and received IP injections of 0.9% NaCl (300 μl per injection, 2 h after TBI and then daily for 14 days); (4) combination of treatments (2) and (3) (“N3 + FO”): mice were fed a diet supplemented with fish oil and received injections of DHA and EPA as described above. The EPA/DHA ratio (70%/30%) in the injectable N3 mixture was dictated by their respective contents in the triple-strength n-3 fish oil (624 mg of EPA and 244 mg of DHA in 1,360 mg of fish oil per capsule).

All experiments were approved by the Institutional Animal Care and Use Committee of the University of Pittsburgh and conducted in accordance with the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals. Specific pathogen free C57BL/6J (B6) and CD-1 mice were purchased from the Jackson Laboratory (Bar Harbor, ME). Mice were habituated to the University of Pittsburgh animal facility for at least one week prior to initiation of experiments. Mice were housed in ventilated caging (Allentown Inc., Allentown, NJ) under 12 h light/dark cycles (0700-1900) and had ad libitum access to food (irradiated 5P76 ProLab IsoPro RMH 3000, [LabDiet, St. Louis, MO]) and water.

All experiments were approved by the Institutional Animal Care and Use Committee of the University of Pittsburgh and conducted in accordance with the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals. C57BL/6J male and female mice used to generate embryos for electroporation and the purchased control group were procured from The Jackson Laboratory (Bar Harbor, ME). CD-1 recipient females and vasectomized males were procured from Charles River Laboratories, Inc. (Wilmington, MA). Mice were housed under 12-h light/dark cycles, with lights on at 7 a.m. and had ad libitum access to food (irradiated 5P76 ProLab IsoProRMH3000; LabDiet, St. Louis, MO) and water.