Differentiated proteome pattern of genetic absence epilepsy rats treated with ketogenic diet.

Objective: The effect of ketogenic diet (KD) on genetic generalized epilepsy has not been specifically reported. Absence epilepsy is a common form of childhood genetic generalized epilepsy. This study aimed to understand the anti-absence effect of the KD on the absence epilepsy in genetic absence epilepsy rats from Strasbourg (GAERS). Methods: 3 Naive GAERS with normal diet and 4 GAERS with KD for 30 days were used in this study. Rats on normal and KD were decapitated, and then frontal cortex and hippocampus were dissected immediately after the brain was removed. The brains were washed in cold 1¥PBS containing protease and phosphatase inhibitors to remove blood and other tissue debris. Brain compartments were pulverized in liquid N2 and homogenized in 50 mM Tris.HCl pH 7.5 buffer including 150 mM NaCl, 1%NP40, 0.2% TritonX100, protease and phosphatase inhibitor cocktails. The homogenate was centrifuged at 16000xg for 15 min at cold, the supernatant was collected and digested as below. Protein from each sample obtained from both tissues was reduced with dithiothreitol (DTT) and alkylated with iodoacetamide (IAA). Proteins were precipitated using methanol/chloroform precipitation protocol. The protein was digested with trypsin (1:100 (w/w, trypsin-LysC/protein). Peptides were run on an EASY-Spray column connected to an Ultimate 3000 RSL nano system (Dionex, Thermo Scientific) in a Q Exactive Plus MS in the data-dependent mode. The MS/MS data were processed using Mascot search engine against the Rat_Protemoe database (29,947 sequences). Localization of identified proteins is predicted with EnrichR tool. Results: As a result, 5 proteins are down-regulated and 6 are up-regulated in the KD-treated GAERS cortex compared to the cortex of GAERS-normal diet. No differentiated protein was detected in the hippocampus of the animals. Conclusion: The differentiated proteins are primarily found in the ketone body metabolism, glycogen metabolism, and trafficking of myristoylated proteins to the cilium. [ABSTRACT FROM AUTHOR]