phenous vein was collected and blood glucose levels were immediately measured using the One Touch II glucose meter. Serum insulin levels were determined using a mouse insulin ELISA kit. Nonesterified fatty acids were determined using a diagnostic reagent kit and triglycerides were determined using a trinder kit. old male mice) using Trizol reagent according to the manufacturer’s protocol and stored at 280uC in 0.1 X SET. To prepare RNA for qRT-PCR, samples were precipitated in 100% ethanol, washed twice with 70% ethanol, then resuspended in autoclaved double-distilled water. Relative levels of PGC-1a, PEPCK, GLUT2, GCK, GAPDH, and G6Pase were determined using TaqMan probe and primer sets designed for each gene, and relative amounts of mRNA were determined based on a standard curve using Stratagene Mx3000P software. Statistical analyses All data are presented as the mean 6 standard error of the mean. Statistical significance was determined by one-way ANOVA and repeated measures ANOVA was used on the glucose tolerance tests followed by Tukey’s Multiple Comparison Test. A p-value of less than 0.05 was considered statistically significant. PrismTM was used to plot data and determine statistical significance. Intraperitoneal glucose tolerance test 10 to 16 week old male mice were injected intraperitoneally with 2.0 mg glucose per gram of body weight using a 20% glucose solution. Blood samples were collected from the hind limb 23472002 saphenous vein at 215, 0, 15, 30, 60 and 120 min after injection and glucose levels were immediately measured. Real-time QRT-PCR One step real time PCR was performed using the Mx3000P and Brilliant QRT-PCR reagents. Total RNA was extracted from frozen mouse livers Comparison of adenosine 39:59-monophosphate-dependent protein kinases from rabbit skeletal and bovine heart muscle. J Biol Chem 250: 7795801. 2. Corbin JD, Sugden PH, West L, Flockhart DA, Lincoln TM, et al. Studies on the properties and mode of action of the purified regulatory subunit of bovine heart adenosine 39:59-monophosphate-dependent protein kinase. J Biol Chem 253: 3997003. 3. Meinkoth JL, Ji Y, Taylor SS, Feramisco JR Dynamics of the distribution of WP 1130 Cyclic AMP-dependent protein kinase in living cells. Proc Natl Acad Sci U S A 87: 9595599. 4. Walsh DA, Van Patten SM Multiple pathway signal transduction by the cAMP-dependent protein kinase. Faseb J 8: 1227236. 5. McKnight GS Cyclic AMP second messenger systems. Curr Opin Cell Biol 3: 21317. 6. Desseyn JL, Burton KA, McKnight GS Expression of a nonmyristylated variant of the catalytic subunit of protein kinase A during male germ-cell development. Proc Natl Acad Sci U S A 97: 6433438. 7. Guthrie CR, Skalhegg BS, McKnight GS Two novel brain-specific splice variants of the murine Cbeta gene of cAMP-dependent protein kinase. J Biol Chem 272: 295609565. 8. Brandon EP, Gerhold KA, Qi M, McKnight GS, Idzerda RL Derivation of novel embryonic stem cell lines and targeting of cyclic AMP-dependent protein kinase genes. Recent Prog Horm Res 50: 40308. 9. Cummings DE, Brandon EP, Planas JV, Motamed K, Idzerda RL, et al. Genetically lean mice result 
from targeted disruption of the RII beta subunit of protein kinase A. Nature 382: 62226. 10. Burton KA, Johnson BD, Hausken ZE, Westenbroek RE, Idzerda RL, et al. Type II regulatory subunits are not required for the anchoring-dependent modulation of Ca2+ channel activity by cAMP-dependent protein kinase. Proc Natl Acad Sci U S A 94: 110671072. 11. Amieux PS, Howe DG, Knicphenous vein was collected and blood glucose levels were immediately measured using the One Touch II glucose meter. Serum insulin levels were determined using a mouse insulin ELISA kit. Nonesterified fatty acids were determined using a diagnostic reagent kit and triglycerides were determined using a trinder kit. old male mice) using Trizol reagent according to the manufacturer’s protocol and stored at 280uC in 0.1 X SET. To prepare RNA for qRT-PCR, samples were precipitated in 100% ethanol, washed twice with 70% ethanol, then resuspended in autoclaved double-distilled water. Relative levels of PGC-1a, PEPCK, GLUT2, GCK, GAPDH, and G6Pase were determined using TaqMan probe and primer sets designed for each gene, and relative amounts of mRNA were determined based on a standard curve using Stratagene Mx3000P software. Statistical analyses All data are presented as the mean 6 standard error of the mean. Statistical significance was determined by one-way ANOVA and repeated measures ANOVA was used on the glucose tolerance tests followed by Tukey’s Multiple Comparison Test. A p-value of less than 0.05 was considered statistically significant. PrismTM was used to plot data and determine statistical significance. Intraperitoneal glucose tolerance test 10 to 16 week old male mice were injected intraperitoneally with 2.0 mg glucose per gram of body weight using a 20% glucose solution. Blood samples were collected from the hind limb saphenous vein at 215, 0, 15, 30, 60 and 120 min after injection and glucose levels were immediately measured. Real-time QRT-PCR One step real time PCR was performed using the Mx3000P and Brilliant QRT-PCR reagents. Total RNA was extracted from frozen mouse livers Comparison of adenosine 39:59-monophosphate-dependent protein kinases from rabbit skeletal and bovine heart muscle. J Biol Chem 250: 7795801. 2. Corbin JD, Sugden PH, West L, Flockhart DA, Lincoln TM, et al. Studies on the properties and mode of action of the purified regulatory subunit of bovine heart adenosine 39:59-monophosphate-dependent protein kinase. J Biol Chem 253: 3997003. 3. Meinkoth JL, Ji Y, Taylor SS, Feramisco JR Dynamics of the distribution of cyclic AMP-dependent protein kinase in living cells. Proc Natl Acad Sci U S A 87: 9595599. 4. Walsh DA, Van Patten SM Multiple pathway signal transduction by the cAMP-dependent protein kinase. Faseb J 8: 1227236. 5. McKnight GS Cyclic AMP second messenger systems. Curr Opin Cell Biol 3: 21317. 6. Desseyn JL, Burton KA, McKnight GS Expression of a nonmyristylated variant of the catalytic subunit of protein kinase A during male germ-cell development. Proc Natl Acad Sci U S A 97: 6433438. 7. Guthrie CR, Skalhegg BS, McKnight GS Two novel brain-specific splice variants of the murine Cbeta gene of cAMP-dependent protein kinase. J Biol Chem 272: 295609565. 8. Brandon EP, Gerhold KA, Qi M, McKnight GS, Idzerda RL Derivation of novel embryonic stem cell lines and targeting of cyclic AMP-dependent protein kinase genes. Recent Prog Horm Res 50: 40308. 9. Cummings DE, Brandon EP, Planas JV, Motamed K, Idzerda RL, et al. Genetically lean mice result from targeted disruption of the RII beta subunit of protein kinase A. Nature 382: 62226. 10. Burton KA, Johnson BD, Hausken ZE, Westenbroek RE, Idzerda RL, et al. Type II regulatory subunits are not required for the anchoring-dependent modulation of Ca2+ channel activity by cAMP-dependent protein kinase. Proc Natl Acad Sci U S A 94: 110671072. 11. Amieux PS, Howe DG, Knic
