Tissue Specific Inhibition of Diet-Induced Thermogenesis

<div>This is a funded MNORC small grant.</div><div><br></div><div>Our objective is to determine the specific roles of white adipose tissue, brown adipose tissue and muscle in diet-induced thermogenesis. We will test the hypothesis that adrenergic signaling in muscle is required for thermogenic adaptations to high fat diet. While our hypothesis focuses on muscle as the central thermogenic organ in response to increased calories, this application will test the roles of white, brown adipose tissue and muscle in mediating the thermogenic/adrenergic response to overnutrition. To do this we will utilize new technologies where  adrenergic signaling can be specifically and acutely ablated in a cell specific manner. We will utilize transgenic, tissue-specific expression of DREADD receptors, coupled to the inhibitory heterotrimeric G proteinGi.</div

Regulation of Hepatic PTG Expression by Glucocorticoids

Grant proposal through the NDSP funding mechanism, sponsored by NURSA. <div><div> </div></div

Cyclic nucleotide binding proteins in the Arabidopsis thaliana and Oryza sativa genomes - Figure 3

<p><strong>Copyright information:</strong></p> <p>Taken from "Cyclic nucleotide binding proteins in the and genomes"</p> <p>BMC Bioinformatics 2005;6():6-6.</p> <p>Published online 11 Jan 2005</p> <p>PMCID:PMC545951.</p> <p>Copyright © 2005 Bridges et al; licensee BioMed Central Ltd.</p> <p>re aligned against several well studied CNB domains including regulatory subunits of PKA (RIα and RIIβ), Epac1, Epac2, and cyclic GMP dependent kinase 2 (CGK2) from humans, HCN2 from mouse and CAP. Highlighted on the alignment are glycine residues involved in loop structures (dark grey arrows), residues forming the hydrophobic pocket for cNMP binding (green arrows) and residues proposed to contact the phosphate of the cNMP (blue arrows). The highly conserved helix capping acidic residue is shown in red. Secondary structure is denoted by arrows above the alignment, with light blue for alpha helices and pink for beta sheets and is based on the secondary structure of HCN2. (B) A homology model of atCNTE1 was generated from the known structures of CNB domains. Key residues are shown as stick representations and are colored and labeled according to the color scheme described in (A). The cGMP ligand is shown in magenta and is based on the structure of cGMP bound to HCN2 [pdb: 1Q3E] superimposed over our model. Figure was generated with Molscript [83] and Raster3D [84].</p

Cyclic nucleotide binding proteins in the Arabidopsis thaliana and Oryza sativa genomes - Figure 2

<p><strong>Copyright information:</strong></p> <p>Taken from "Cyclic nucleotide binding proteins in the and genomes"</p> <p>BMC Bioinformatics 2005;6():6-6.</p> <p>Published online 11 Jan 2005</p> <p>PMCID:PMC545951.</p> <p>Copyright © 2005 Bridges et al; licensee BioMed Central Ltd.</p> <p>re aligned against several well studied CNB domains including regulatory subunits of PKA (RIα and RIIβ), Epac1, Epac2, and cyclic GMP dependent kinase 2 (CGK2) from humans, HCN2 from mouse and CAP. Highlighted on the alignment are glycine residues involved in loop structures (dark grey arrows), residues forming the hydrophobic pocket for cNMP binding (green arrows) and residues proposed to contact the phosphate of the cNMP (blue arrows). The highly conserved helix capping acidic residue is shown in red. Secondary structure is denoted by arrows above the alignment, with light blue for alpha helices and pink for beta sheets and is based on the secondary structure of HCN2. (B) A homology model of atCNTE1 was generated from the known structures of CNB domains. Key residues are shown as stick representations and are colored and labeled according to the color scheme described in (A). The cGMP ligand is shown in magenta and is based on the structure of cGMP bound to HCN2 [pdb: 1Q3E] superimposed over our model. Figure was generated with Molscript [83] and Raster3D [84].</p

Cyclic nucleotide binding proteins in the Arabidopsis thaliana and Oryza sativa genomes - Figure 4

<p><strong>Copyright information:</strong></p> <p>Taken from "Cyclic nucleotide binding proteins in the <em>Arabidopsis thaliana</em> and <em>Oryza sativa</em> genomes"</p> <p>BMC Bioinformatics 2005;6():6-6.</p> <p>Published online 11 Jan 2005</p> <p>PMCID:PMC545951.</p> <p>Copyright © 2005 Bridges et al; licensee BioMed Central Ltd.</p> <p><strong>Biochemical evidence for lack of a cyclic nucleotide dependent kinase in <em>Arabidopsis thaliana</em></strong>. (A) Protein kinase assays using Kemptide as a substrate. Assays were conducted on identically prepared extracts of Arabidopsis and rat adipose tissue in the presence or absence (control) of 10 μM cyclic nucleotide as indicated. Scale is offset in order to visualize both sets of results. All assays were performed in duplicate from three separate preparations and error bars indicate standard error for three separate preparations. (B) Western blotting of extracts with PKA catalytic (PKAcs) and regulatory (RII) subunit polyclonal antibodies. The PKAcs antibody was affinity purified according to [82] and used at 0.5 μg/mL while the RII antibody was used as crude serum at 5000X dilution. Lanes are as follows (A), 10 ng of purified bovine PKAcs or RII, (B) 25 μg clarified crude Arabidopsis extract, (C) 25 μg clarified crude rat adipocyte extract. Positions of mammalian PKA and RII are indicated with arrows.</p

The Role of dTORC1 on Muscle Development, Function and Longevity in Drosophila

<p>The TORC1 signaling pathway is critical for cell growth and proliferation. It has been implicated in disorders ranging from diabetes and obesity to depression and cancer. Previous work has implicated the TORC1 pathway in the regulation of longevity and muscle function in a variety of model systems. In this study, we manipulated the activity of mTORC1 in muscle tissue by using the Drosophila GAL4/UAS system. We did this by knocking down both positive (Raptor) and negative (Tsc1) regulators of dTORC1 function in both cardiac and skeletal muscles. We observed that genetic inhibition of TORC1 in skeletal but not cardiac muscle leads to reduced viability using the skeletal muscle GAL4 drivers (C179-GAL4 and 24B-GAL4). Using climbing assays, we have also examined the effects of these manipulations on muscle function and have observed reduced y motility with both Raptor and Tsc1 inhibition in muscle. We found that activation of TORC1 in y skeletal muscle tissue also leads to signicant reductions in lifespan. Both the reduced muscle function and shortened lifespan are consistent with results obtained in a mouse model of muscle Tsc1 deletion. Expression of both positive and negative regulators of TORC1 specically in cardiac muscle using the Hand-GAL4 driver had no dramatic effects on either viability or longevity. These data provide insights into the role of muscle TORC1 activity in development, muscle function and longevity.</p