Advertisement

Diabetologia

, Volume 61, Issue 8, pp 1849–1855 | Cite as

Skeletal muscle-specific Cre recombinase expression, controlled by the human α-skeletal actin promoter, improves glucose tolerance in mice fed a high-fat diet

  • Rami Al Batran
  • Keshav Gopal
  • Mackenzie D. Martin
  • Kim L. Ho
  • Malak Almutairi
  • Hanin Aburasayn
  • Farah Eaton
  • Jonathan E. Campbell
  • John R. Ussher
Short Communication

Abstract

Aims/hypothesis

Cre-loxP systems are frequently used in mouse genetics as research tools for studying tissue-specific functions of numerous genes/proteins. However, the expression of Cre recombinase in a tissue-specific manner often produces undesirable changes in mouse biology that can confound data interpretation when using these tools to generate tissue-specific gene knockout mice. Our objective was to characterise the actions of Cre recombinase in skeletal muscle, and we anticipated that skeletal muscle-specific Cre recombinase expression driven by the human α-skeletal actin (HSA) promoter would influence glucose homeostasis.

Methods

Eight-week-old HSA-Cre expressing mice and their wild-type littermates were fed a low- or high-fat diet for 12 weeks. Glucose homeostasis (glucose/insulin tolerance testing) and whole-body energy metabolism (indirect calorimetry) were assessed. We also measured circulating insulin levels and the muscle expression of key regulators of energy metabolism.

Results

Whereas tamoxifen-treated HSA-Cre mice fed a low-fat diet exhibited no alterations in glucose homeostasis, we observed marked improvements in glucose tolerance in tamoxifen-treated, but not corn-oil-treated, HSA-Cre mice fed a high-fat diet vs their wild-type littermates. Moreover, Cre dissociation from heat shock protein 90 and translocation to the nucleus was only seen following tamoxifen treatment. These improvements in glucose tolerance were not due to improvements in insulin sensitivity/signalling or enhanced energy metabolism, but appeared to stem from increases in circulating insulin.

Conclusions/interpretation

The intrinsic glycaemia phenotype in the HSA-Cre mouse necessitates the use of HSA-Cre controls, treated with tamoxifen, when using Cre-loxP models to investigate skeletal muscle-specific gene/protein function and glucose homeostasis.

Keywords

Cre recombinase Glucose tolerance Insulin Skeletal muscle 

Abbreviations

BSA

Bovine serum albumin

CM

Conditioned media

D-Ala2 GIP

D-Ala2 glucose-dependent insulinotropic polypeptide

GSIS

Glucose-stimulated insulin secretion

GSK

Glycogen synthase kinase

HFD

High-fat diet

HSA

Human α-skeletal actin

Hsp

Heat shock protein

KO

Knockout

LFD

Low-fat diet

MCK

Muscle creatine kinase

mTOR

Mammalian target of rapamycin

PPAR

Peroxisome proliferator-activated receptor

TAG

Triacylglycerol

VDAC

Voltage-dependent anion-selective channel

WT

Wild-type

Notes

Acknowledgements

We thank J. Kruger (Health Sciences Laboratory Animal Services, University of Alberta) for husbandry and maintenance of our HSA-Cre mouse colony.

Contribution statement

RAB, JEC and JRU were involved with conception and design of the study. RAB, KG, MDM, KLH, MA, HA and FE were involved with data acquisition. RAB, KG and MDM were involved with data analysis and interpretation. RAB and JRU drafted the manuscript. All authors contributed to critically revising the article for important intellectual content and gave their final approval of the version to be published. JRU is the guarantor of this work.

Funding

These studies were supported in part by a discovery grant (RGPIN 04946) from the Natural Sciences and Engineering Research Council of Canada to JRU. JRU is a Scholar of the Canadian Diabetes Association and a New Investigator of the Heart and Stroke Foundation of Alberta, NWT & Nunavut. RAB is a postdoctoral fellow of the Canadian Institutes of Health Research and the Canadian Diabetes Association.

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Supplementary material

125_2018_4643_MOESM1_ESM.pdf (30 kb)
ESM Table 1 (PDF 29 kb)

References

  1. 1.
    Glaser S, Anastassiadis K, Stewart AF (2005) Current issues in mouse genome engineering. Nat Genet 37:1187–1193CrossRefPubMedGoogle Scholar
  2. 2.
    Oropeza D, Jouvet N, Budry L et al (2015) Phenotypic characterization of MIP-CreERT1Lphi mice with transgene-driven islet expression of human growth hormone. Diabetes 64:3798–3807CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Koitabashi N, Bedja D, Zaiman AL et al (2009) Avoidance of transient cardiomyopathy in cardiomyocyte-targeted tamoxifen-induced MerCreMer gene deletion models. Circ Res 105:12–15CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Ussher JR, Jaswal JS, Wagg CS et al (2009) Role of the atypical protein kinase Czeta in regulation of 5'-AMP-activated protein kinase in cardiac and skeletal muscle. Am J Physiol Endocrinol Metab 297:E349–E357CrossRefPubMedGoogle Scholar
  5. 5.
    Hohmeier HE, Mulder H, Chen G, Henkel-Rieger R, Prentki M, Newgard CB (2000) Isolation of INS-1-derived cell lines with robust ATP-sensitive K+ channel-dependent and -independent glucose-stimulated insulin secretion. Diabetes 49:424–430CrossRefPubMedGoogle Scholar
  6. 6.
    Ussher JR, Fillmore N, Keung W et al (2016) Genetic and pharmacological inhibition of malonyl CoA decarboxylase does not exacerbate age-related insulin resistance in mice. Diabetes 65:1883–1891CrossRefPubMedGoogle Scholar
  7. 7.
    Ussher JR, Baggio LL, Campbell JE et al (2014) Inactivation of the cardiomyocyte glucagon-like peptide-1 receptor (GLP-1R) unmasks cardiomyocyte-independent GLP-1R-mediated cardioprotection. Mol Metab 3:507–517CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Kitamura T, Kitamura Y, Nakae J et al (2004) Mosaic analysis of insulin receptor function. J Clin Invest 113:209–219CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Gopal K, Saleme B, Al Batran R et al (2017) FoxO1 regulates myocardial glucose oxidation rates via transcriptional control of pyruvate dehydrogenase kinase 4 expression. Am J Physiol Heart Circ Physiol 313:H479–H490CrossRefPubMedGoogle Scholar
  10. 10.
    Muoio DM, Neufer PD (2012) Lipid-induced mitochondrial stress and insulin action in muscle. Cell Metab 15:595–605CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Vandanmagsar B, Warfel JD, Wicks SE et al (2016) Impaired mitochondrial fat oxidation induces FGF21 in muscle. Cell Rep 15:1686–1699CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Holt LJ, Brandon AE, Small L et al (2018) Ablation of Grb10 specifically in muscle impacts muscle size and glucose metabolism in mice. Endocrinology 159:1339–1351CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Rami Al Batran
    • 1
    • 2
    • 3
  • Keshav Gopal
    • 1
    • 2
    • 3
  • Mackenzie D. Martin
    • 4
  • Kim L. Ho
    • 1
    • 2
    • 3
  • Malak Almutairi
    • 1
    • 2
    • 3
  • Hanin Aburasayn
    • 1
    • 2
    • 3
  • Farah Eaton
    • 1
    • 2
    • 3
  • Jonathan E. Campbell
    • 4
  • John R. Ussher
    • 1
    • 2
    • 3
  1. 1.Katz Centre for Pharmacy and Health Research, Faculty of Pharmacy and Pharmaceutical SciencesUniversity of AlbertaEdmontonCanada
  2. 2.Alberta Diabetes InstituteUniversity of AlbertaEdmontonCanada
  3. 3.Mazankowski Alberta Heart InstituteUniversity of AlbertaEdmontonCanada
  4. 4.Duke Molecular Physiology InstituteDuke UniversityDurhamUSA

Personalised recommendations