MicehandledwiththesameprotocolasdepictedinFig.5B.pptVIP

Mice handled with the same protocol as depicted in Fig. 5B were evaluated 2 weeks 2w after treatment. Joseane Morari et al. Diabetes 2014;63:3770-3784 ?2014 by American Diabetes Association Mice handled with the same protocol as depicted in Fig. 5B were evaluated 2 weeks 2w after treatment. A: Hypothalami were obtained for RNA extraction. The cDNA that was produced from total RNA was used in real-time PCR assays to determine the expression of fractalkine CX3CL1 A . Body mass was determined weekly B ; body mass gain was determined at the end of the experimental period C , as was body mass composition by densitometric scanning: fat mass D and lean mass E . At the end of the experimental period, randomly selected mice were submitted to either a glucose tolerance test, where the area under the glucose curve AUC was calculated F , or an insulin tolerance test, where the constant for glucose decay kITT was calculated G . H–J: Hypothalami were obtained for RNA extraction. The cDNA that was produced from total RNA was used in real-time PCR assays to determine the expression of fractalkine receptor CX3CR1 H , TNFα I , and IL1β J . K: The protocol that was used to treat male Swiss mice with a bilateral intracerebral injection in the arcuate nucleus 2.0 μL, 40 pmol/L, each side of SI or SC against fractalkine; after SI or SC injections, mice were assigned an HFD for 2 weeks 2w , and gene expression of fractalkine CX3CL1 was determined by real-time PCR in the medium-basal hypothalamus MBH L and in the frontal cortex FC M ; some mice were randomly selected for a glucose tolerance test, where the area under the glucose curve AUC was calculated N . In real-time PCR experiments, the results are expressed as the relative gene expression. In all experiments, n 6: #P 0.05 compared with SC.