Introduction: We observed that two-lines of transgenic mice ubiquitously expressing Spinacia microsomal fad2 gene for omega-6 desaturase were lean phenotype. These mice were shown to be resistant to obesity induced by high fat diet, indicating their basal metabolic rate is also elevated. Here, we investigated about differentially expressed proteins between two-lines of transgenic and wild-type mice.
Methods: Two mg of white adipose tissue and 1mg of skeletal muscle were homogenized and added 4uL of lysis buffer, containing 7M urea, 2M thiourea, 4% CHAPS, 0.5%IPG buffer pH 3-11 nonlinear,0.05%TBP and protease inhibitor cocktail. Proteins were separated by 2-DE and were visualized using SYPRO Ruby. The separated proteins were identified by MALDI-TOF/TOF and Mascot search (Matrix science). Expression levels of the separated proteins calculated with PG220 and TT900 software (Nonlinear dynamics). The statistical analysis of quantitative expression values of each protein was done between two lines of transgenic and wild type mice.
Result: In the white adipose tissue and skeletal muscle, 810 and 635 protein spots were commonly detected on 2-DE gels and 329 and 315 protein spots were identified by MALDI-TOF/TOF, respectively. When two lines of transgenic and wild-type mice were compared in the white adipose tissue, we observed that 38 protein spots were differentially expressed. These included glycolysis-related proteins. Moreover, 30 proteins spots were shown to be differential expressed in comparison between skeletal muscle from two lines of transgenic and wild-type mice. These proteins were involving in glycolysis and mitochondrial function. The current data suggest that up-regulated energy metabolism may induce lean phenotype in transgenic mice bearing fad2 gene.
Acknowledgement: This work was supported by grant from the Wakayama Prefecture Collaboration of Regional Entities for the Advancement of Technological Excellence of JST.
