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Skeletal muscle–specific ATGL deletion and its impact on lipid homeostasis ...
Published: 17 September 2013
FIG. 2. Skeletal muscle–specific ATGL deletion and its impact on lipid homeostasis in SMAKO mice. A: The LoxP-modified ATGL construct. B: ATGL mRNA expression relative to 18S control gene by quantitative PCR in muscle and nonmuscle tissues with endogenous ATGL expression in quadriceps arbitrarily set to 1 (♀, 10 weeks, chow, fasted 12 h; n = 5 to 6/group). Percent decrease in ATGL mRNA expression in SMAKO relative to WT mice for select muscle tissues is shown in the table (bottom). C: ATGL protein expression relative to Ran GTPase (RAN) control in skeletal versus cardiac muscle (♀, 10 weeks, chow, fasted 12 h, gastrocnemius and heart; n = 5 to 6/group). D: TAG hydrolase activity at baseline and in the presence of the HSL-specific inhibitor 76-0079, the ATGL-specific activator CGI-58, or HSL inhibitor plus CGI-58 (♂, 28 weeks, chow, fasted 12 h, red gastrocnemius; n = 6/group). E: Skeletal muscle histology of control (left), SMAKO (middle), and GAKO (right) mice including general morphology by H&E staining (top) and IMTG content by ORO staining (bottom) (♂, 28 weeks, chow, fasted 12 h, gastrocnemius-plantaris-soleus [GPS] complex). F: IMTG content by ORO staining using quantitative IF (♂, 28 weeks, fasted 12 h, GPS complex, average of four muscle areas each; n = 4/group). Type 2B fibers of chow-fed WT mice are arbitrarily set to 1. G: Intramyocellular DAG, ceramide, and FA-CoA content using biochemical analysis of whole muscle (♂, 28 weeks, fasted 12 h, quadriceps; n = 3 to 4/group). For overall effects having P < 0.05: D, diet; F, fiber type; G, genotype; T, treatment (with HSL-inhibitor or CGI-58). For specific comparisons having P < 0.05: #for effect of diet; *for effect of genotype; and @for effect of treatment. AU, arbitrary units; BAC, bacterial artificial chromosome; BAT, brown adipose tissue; D, diet; EDL, extensor digitorum longus; ES, embryonic stem; Gas, gastrocnemius; Hrt, heart; PGAT, perigonadal adipose tissue; Quad, quadriceps; Sol, soleus; TA, tibialis anterior. FIG. 2. Skeletal muscle–specific ATGL deletion and its impact on lipid homeostasis in SMAKO mice. A: The LoxP-modified ATGL construct. B: ATGL mRNA expression relative to 18S control gene by quantitative PCR in muscle and nonmuscle tissues with endogenous ATGL expression in quadriceps arbitrarily set to 1 (♀, 10 weeks, chow, fasted 12 h; n = 5 to 6/group). Percent decrease in ATGL mRNA expression in SMAKO relative to WT mice for select muscle tissues is shown in the table (bottom). C: ATGL protein expression relative to Ran GTPase (RAN) control in skeletal versus cardiac muscle (♀, 10 weeks, chow, fasted 12 h, gastrocnemius and heart; n = 5 to 6/group). D: TAG hydrolase activity at baseline and in the presence of the HSL-specific inhibitor 76-0079, the ATGL-specific activator CGI-58, or HSL inhibitor plus CGI-58 (♂, 28 weeks, chow, fasted 12 h, red gastrocnemius; n = 6/group). E: Skeletal muscle histology of control (left), SMAKO (middle), and GAKO (right) mice including general morphology by H&E staining (top) and IMTG content by ORO staining (bottom) (♂, 28 weeks, chow, fasted 12 h, gastrocnemius-plantaris-soleus [GPS] complex). F: IMTG content by ORO staining using quantitative IF (♂, 28 weeks, fasted 12 h, GPS complex, average of four muscle areas each; n = 4/group). Type 2B fibers of chow-fed WT mice are arbitrarily set to 1. G: Intramyocellular DAG, ceramide, and FA-CoA content using biochemical analysis of whole muscle (♂, 28 weeks, fasted 12 h, quadriceps; n = 3 to 4/group). For overall effects having P < 0.05: D, diet; F, fiber type; G, genotype; T, treatment (with HSL-inhibitor or CGI-58). For specific comparisons having P < 0.05: #for effect of diet; *for effect of genotype; and @for effect of treatment. AU, arbitrary units; BAC, bacterial artificial chromosome; BAT, brown adipose tissue; D, diet; EDL, extensor digitorum longus; ES, embryonic stem; Gas, gastrocnemius; Hrt, heart; PGAT, perigonadal adipose tissue; Quad, quadriceps; Sol, soleus; TA, tibialis anterior. More
Meeting Abstracts
Journal: Diabetes
Diabetes 2000;49(3):492–499
Published: 01 March 2000
..., and GATA52A04) was determined by contains multiple susceptibility genes (3 11). IDDM2 has sequence tagged site content mapping of bacterial arti- been identi ed as the variable number of tandem repeats ficial chromosome (BAC) and yeast artificial chromo- immediately 5 to the insulin gene (12 19), which appears...
Journal Articles
Journal: Diabetes
Diabetes 2011;60(3):775–783
Published: 21 February 2011
... in inflammatory states, we generated mice lacking murine resistin but transgenic for a bacterial artificial chromosome containing human resistin (BAC-Retn), whose expression was similar to that in humans. The metabolic and molecular phenotypes of BAC-Retn mice were assessed after acute...
Includes: Supplementary data
Meeting Abstracts
Journal: Diabetes
Diabetes 2000;49(3):508–512
Published: 01 March 2000
... at 7419 and 137 were in LD with D6S281/AFM234 (Table 2). Indeed, LD was strong, Additional information can be found in an online appendix at www. even between D6S281 and 137 (D , 0.63; 2, 89.6), a physical diabetes.org/diabetes/appendix.asp. distance of ~142 kb. BAC, bacterial artificial chromosome; EST...
Journal Articles
Journal: Diabetes
Diabetes 2006;55(7):2153–2156
Published: 01 July 2006
... requests to Professor Roger D. Cox, Medical Research Council, Mammalian Genetics Unit, Harwell, Oxfordshire, OX11 0RD, U.K. E-mail: r.cox@har.mrc.ac.uk 18 4 2006 17 3 2006 DIABETES 2006 BAC, bacterial artificial chromosome IPGTT, intraperitoneal glucose tolerance test Nnt...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2002;51(7):2334–2336
Published: 01 July 2002
... January 2002 and accepted in revised form 16 April 2002. BAC, bacterial artificial chromosome; IFN-γ, interferon-γ; IL, interleukin; LPS, lipopolysaccharides; SNP, single-nucleotide polymorphism; TDT, transmission disequilibrium test; UTR, untranslated region. The microsatellite marker (ATT)n...
Journal Articles
Journal: Diabetes
Diabetes 2003;52(6):1551–1556
Published: 01 June 2003
... of an Idd10 B6 bacterial artificial chromosome (BAC) clone contig and the ascertainment of its gene content. Consequently, the Idd10 interval was more precisely defined by fine mapping the recombination break points of key congenic strains using polymorphic microsatellite markers obtained from...
Journal Articles
Journal: Diabetes
Diabetes 1997;46(6):1081–1086
Published: 01 June 1997
... gene. 3 BAC, bacterial artifical chromosome; HNF-1α, hepatocyte nuyclear factor-1α; MODY, maturity-onset diabetes of the young; PAC, P1 artificial chromosome; PCR, polymerase chain reaction; SSCP, single-strand conformation polymorphism. 12 2 1997 10 3 1997 10 3 1997...
Journal Articles
Journal: Diabetes
Diabetes 2002;51(11):3318–3325
Published: 01 November 2002
... in subsequent rounds of library screening. Additional contigs were initiated by library screening with the following microsatellites: D1S1644, D1S439, D1S1656, and D1S2712. Bacterial artificial chromosome (BAC) clones (n = 104) were isolated by screening high...
Journal Articles
Journal: Diabetes
Diabetes 2006;55(12):3366–3371
Published: 01 December 2006
... for the role of the neurotrophin BDNF in human energy homeostasis, as well as in cognitive function, memory, and behavior. PCR was used to generate ∼500 bp probes mapping to restriction fragments of the genomic region of interest, using bacterial artificial chromosome (BAC)-derived miniprep DNA...
Includes: Supplementary data
Meeting Abstracts
Journal: Diabetes
Diabetes 1999;48(10):2099–2102
Published: 01 October 1999
... The nucleotide sequences reported in this paper have been submitted to were 231 diabetic and 152 nondiabetic family members. The GenBank with accession numbers Z49826, AF133253, and AF133493-504. Additional information can be found in an online appendix at www. diabetes.org/diabetes/appendix.htm. BAC, bacterial...
Meeting Abstracts
Journal: Diabetes
Diabetes 1999;48(9):1877–1880
Published: 01 September 1999
... pre- 19 May 1999. vious analysis with antisense probes directed against the 3 -end of the mRNA (5) and shows that R5020 regulation of BAC, bacterial artificial chromosome; GRE, glucocorticoid response IRS-2 mRNA occurs at the transcriptional level. element; h, human; IRS, insulin receptor substrate...
Journal Articles
Journal: Diabetes
Diabetes 2002;51(11):3342–3346
Published: 01 November 2002
...), but the strength of the association was not sufficient to account for the 1q linkage in the Pimas ( 12 ). Using BLAST searches of GenBank, we identified three bacterial artificial chromosomes (BACs) (201L5, 536C5, and 2338N5) containing portions of the KCNJ10 locus. We determined that this gene spans...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2005;54(4):1179–1184
Published: 01 April 2005
... is found at highest levels in pancreas and small intestine ( 6 ). BAC, bacterial artificial chromosome EGF, epidermal growth factor SNP, single nucleotide polymorphism SSCP, single-strand conformational polymorphism UT, untranslated region We genotyped African-American and Caucasian...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2008;57(1):133–141
Published: 01 January 2008
... 2008 BAC, bacterial artificial chromosome EGFP, enhanced green fluorescent protein FRT, FLP recombination target G6P, glucose-6-phosphate G6PC, glucose-6-phosphatase catalytic subunit IGRP, islet-specific G6PC-related protein IRES, internal ribosome entry site TSS, transcription start site...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2002;51(3):591–598
Published: 01 March 2002
... DNA sequencing service managed by Dr. Nick Helps (www.dnaseq.co.uk). The human clone, which contained 200 bp that were highly similar to nucleotide (nt) 1–200 of rat GL, was used in a hybridization screen of a human Bacterial Artificial Chromosome (BAC) library (performed by Genome Systems...
Journal Articles
Journal: Diabetes
Diabetes 2004;53(5):1375–1384
Published: 01 May 2004
... to Alessandro Doria, MD, PhD, Section on Genetics & Epidemiology, Joslin Diabetes Center, One Joslin Place, Boston, MA 02215. E-mail: alessandro.doria@joslin.harvard.edu 29 1 2004 10 11 2003 DIABETES 2004 BAC, bacterial artificial chromosome CEPH, Centre d’Etude du Polymorphisme...
Journal Articles
Journal: Diabetes
Diabetes 2005;54(11):3319–3325
Published: 01 November 2005
... DIABETES 2005 Primers were designed to amplify and sequence exons B-6 from the PPARG genomic sequence (NCBI accession no. AY157024), exons A1–A2 from bacterial artificial chromosome (BAC) clone 335I9 (AC091492), the γ2 promoter from BAC clone 30G23 (AC090947), and the 3′ flanking sequence...
Journal Articles
Journal: Diabetes
Diabetes 2014;63(4):1182–1184
Published: 13 March 2014
... for homology regions. iCre stands for improved Cre, where prokaryotic codons are humanized and CpG islands are minimized. Rosa26-tdRFP is a red fluorescent protein knock-in to Rosa26 locus in the mouse. Richards et al. generated GLP1R transgenic mice using a bacterial artificial chromosome (BAC)-based...
Journal Articles
Journal: Diabetes
Diabetes 2014;63(2):815
Published: 16 January 2014
... record. The authors have replicated the results of the original figure using their remaining bacterial artificial chromosome (BAC) mouse line (see Fig. 1C replacement below) and have also previously replicated the rescue of the glucose tolerance phenotype in this line, thus showing...