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Animal Models

Neuronal growth regulator 1-deficient mice show increased adiposity and decreased muscle mass

International Journal of Obesity volume 43, pages 1769–1782 (2019)Cite this article

Subjects

Abstract

Background

Neuronal growth regulator 1 (NEGR1) is a glycosylphosphatidylinositol-anchored membrane protein that mediates neural cell communication and synapse formation. Multiple genome-wide association studies have reported that variations in NEGR1 are associated with human body weight control. Recently, we found that NEGR1 is involved in intracellular cholesterol trafficking, suggesting that it performs a non-central nervous system (CNS) function associated with human obesity.

Methods

We compared peripheral tissues such as the adipose, liver, and skeletal muscle tissues of Negr1−/− and Negr1+/+ (wild-type [WT]) C57BL/6 mice (n = 5–14). Intracellular lipid content was measured, and lipid accumulation was visualized by staining tissue cross-sections with lipid-specific stains. Muscle capacity of the WT and Negr1−/− mice was determined by performing a treadmill endurance test, and muscle fiber size was examined. Plasma glucose and insulin levels were measured, and glucose and insulin tolerance tests were performed.

Results

The Negr1−/− mice showed a significant increase in fat mass (~1.5-fold increase in the epididymal white adipose tissue, p = 0.000002), with abnormally enlarged adipose cells, compared with the WT mice. Primary adipocytes of the Negr1−/− mice contained enlarged cytosolic lipid droplets (p = 0.049). Moreover, these mice showed significant hepatic lipid accumulation (~2.3-fold increase, p = 0.043). Although the Negr1−/− mice did not show a significant change in plasma lipoprotein level, they showed a >1.3-fold increase in a serum glucose (p = 0.0002) and insulin (p = 0.016) levels. Moreover, the Negr1−/− mice showed decreased muscle capacity, as indicated by a decrease in muscle mass (p = 0.000003).

Conclusion

These results indicate that NEGR1 deficiency induces abnormal fat deposition in various peripheral cells, especially fat and liver tissue cells, and suggest that NEGR1 is a potential molecular target for designing anti-obesity drugs to regulate body weight both centrally and peripherally.

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References

  1. Funatsu N, Miyata S, Kumanogoh H, Shigeta M, Hamada K, Endo Y, et al. Characterization of a novel rat brain glycosylphosphatidylinositol-anchored protein (Kilon), a member of the IgLON cell adhesion molecule family. J Biol Chem. 1999;274:8224–30.

    Article  CAS  Google Scholar 

  2. Hashimoto T, Maekawa S, Miyata S. IgLON cell adhesion molecules regulate synaptogenesis in hippocampal neurons. Cell Biochem Funct. 2009;27:496–8.

    Article  CAS  Google Scholar 

  3. Kim H, Hwang JS, Lee B, Hong J, Lee S. Newly Identified Cancer-Associated Role of Human Neuronal Growth Regulator 1 (NEGR1). J Cancer. 2014;5:598–608.

    Article  CAS  Google Scholar 

  4. Willer CJ, Speliotes EK, Loos RJ, Li S, Lindgren CM, Heid IM, et al. Six new loci associated with body mass index highlight a neuronal influence on body weight regulation. Nat Genet. 2009;41:25–34.

    Article  CAS  Google Scholar 

  5. Wheeler E, Huang N, Bochukova EG, Keogh JM, Lindsay S, Garg S, et al. Genome-wide SNP and CNV analysis identifies common and low-frequency variants associated with severe early-onset obesity. Nat Genet. 2013;45:513–7.

    Article  CAS  Google Scholar 

  6. Lee AW, Hengstler H, Schwald K, Berriel-Diaz M, Loreth D, Kirsch M, et al. Functional inactivation of the genome-wide association study obesity gene neuronal growth regulator 1 in mice causes a body mass phenotype. PLoS ONE. 2012;7:e41537.

    Article  CAS  Google Scholar 

  7. Boender AJ, van Gestel MA, Garner KM, Luijendijk MC, Adan RA. The obesity-associated gene Negr1 regulates aspects of energy balance in rat hypothalamic areas. Physiological reports. 2014;2:e12083.

    Article  Google Scholar 

  8. Kim H, Chun Y, Che L, Kim J, Lee S, Lee S. The new obesity-associated protein, neuronal growth regulator 1 (NEGR1), is implicated in Niemann-Pick disease Type C (NPC2)-mediated cholesterol trafficking. Biochem Biophys Res Commun. 2017;482:1367–74.

    Article  CAS  Google Scholar 

  9. Chun Y, Kim R, Lee S. Centromere Protein (CENP)-W interacts with Heterogeneous Nuclear Ribonucleoprotein (hnRNP) U and may contribute to kinetochore-microtubule attachment in mitotic cells. PLoS ONE. 2016;11:e0149127.

    Article  Google Scholar 

  10. Fisher FM, Kleiner S, Douris N, Fox EC, Mepani RJ, Verdeguer F, et al. FGF21 regulates PGC-1alpha and browning of white adipose tissues in adaptive thermogenesis. Genes Dev. 2012;26:271–81.

    Article  CAS  Google Scholar 

  11. Handschin C, Chin S, Li P, Liu F, Maratos-Flier E, Lebrasseur NK, et al. Skeletal muscle fiber-type switching, exercise intolerance, and myopathy in PGC-1alpha muscle-specific knock-out animals. J Biol Chem. 2007;282:30014–21.

    Article  CAS  Google Scholar 

  12. Lefterova MI, Zhang Y, Steger DJ, Schupp M, Schug J, Cristancho A, et al. PPARgamma and C/EBP factors orchestrate adipocyte biology via adjacent binding on a genome-wide scale. Genes Dev. 2008;22:2941–52.

    Article  CAS  Google Scholar 

  13. Beller M, Thiel K, Thul PJ, Jackle H. Lipid droplets: a dynamic organelle moves into focus. FEBS Lett. 2010;584:2176–82.

    Article  CAS  Google Scholar 

  14. Morrison S, McGee SL. 3T3-L1 adipocytes display phenotypic characteristics of multiple adipocyte lineages. Adipocyte. 2015;4:295–302.

    Article  CAS  Google Scholar 

  15. Ducharme NA, Bickel PE. Lipid droplets in lipogenesis and lipolysis. Endocrinology. 2008;149:942–9.

    Article  CAS  Google Scholar 

  16. Bechmann LP, Hannivoort RA, Gerken G, Hotamisligil GS, Trauner M, Canbay A. The interaction of hepatic lipid and glucose metabolism in liver diseases. J Hepatol. 2012;56:952–64.

    Article  CAS  Google Scholar 

  17. Wendel AA, Lewin TM, Coleman RA. Glycerol-3-phosphate acyltransferases: rate limiting enzymes of triacylglycerol biosynthesis. Biochim Biophys Acta. 2009;1791:501–6.

    Article  CAS  Google Scholar 

  18. Lee SJ, Kang JH, Iqbal W, Kwon OS. Proteomic analysis of mice fed methionine and choline deficient diet reveals marker proteins associated with steatohepatitis. PLoS ONE. 2015;10:e0120577.

    Article  Google Scholar 

  19. Coleman SK, Rebalka IA, D’Souza DM, Hawke TJ. Skeletal muscle as a therapeutic target for delaying type 1 diabetic complications. World J Diabetes. 2015;6:1323–36.

    Article  Google Scholar 

  20. Sutton AK, Myers MG Jr., Olson DP. The Role of PVH Circuits in Leptin Action and Energy Balance. Annu Rev Physiol. 2016;78:207–21.

    Article  CAS  Google Scholar 

  21. Yan X, Wang Z, Schmidt V, Gauert A, Willnow TE, Heinig M, et al. Cadm2 regulates body weight and energy homeostasis in mice. Mol Metab. 2018;8:180–8.

    Article  CAS  Google Scholar 

  22. Yang HJ, Xia YY, Wang L, Liu R, Goh KJ, Ju PJ, et al. A novel role for neural cell adhesion molecule in modulating insulin signaling and adipocyte differentiation of mouse mesenchymal stem cells. J Cell Sci. 2011;124(Pt 15):2552–60.

    Article  Google Scholar 

  23. Miyata S, Taguchi K, Maekawa S. Dendrite-associated opioid-binding cell adhesion molecule localizes at neurosecretory granules in the hypothalamic magnocellular neurons. Neuroscience. 2003;122:169–81.

    Article  CAS  Google Scholar 

  24. Miyata S, Matsumoto N, Taguchi K, Akagi A, Iino T, Funatsu N, et al. Biochemical and ultrastructural analyses of IgLON cell adhesion molecules, Kilon and OBCAM in the rat brain. Neuroscience. 2003;117:645–58.

    Article  CAS  Google Scholar 

  25. Speakman JR. Functional analysis of seven genes linked to body mass index and adiposity by genome-wide association studies: a review. Hum Hered. 2013;75:57–79.

    Article  CAS  Google Scholar 

  26. Simon MM, Greenaway S, White JK, Fuchs H, Gailus-Durner V, Wells S, et al. A comparative phenotypic and genomic analysis of C57BL/6J and C57BL/6N mouse strains. Genome Biol. 2013;14:R82.

    Article  Google Scholar 

  27. Fisher-Wellman KH, Ryan TE, Smith CD, Gilliam LA, Lin CT, Reese LR, et al. A direct comparison of metabolic responses to high-fat diet in C57BL/6J and C57BL/6NJ mice. Diabetes. 2016;65:3249–61.

    Article  CAS  Google Scholar 

  28. Pol A, Gross SP, Parton RG. Review: biogenesis of the multifunctional lipid droplet: lipids, proteins, and sites. J Cell Biol. 2014;204:635–46.

    Article  CAS  Google Scholar 

  29. Wilfling F, Haas JT, Walther TC, Farese RV Jr. Lipid droplet biogenesis. Curr Opin Cell Biol. 2014;29:39–45.

    Article  CAS  Google Scholar 

  30. Jacquier N, Choudhary V, Mari M, Toulmay A, Reggiori F, Schneiter R. Lipid droplets are functionally connected to the endoplasmic reticulum in Saccharomyces cerevisiae. J Cell Sci. 2011;124(Pt 14):2424–37.

    Article  CAS  Google Scholar 

  31. Mishra S, Khaddaj R, Cottier S, Stradalova V, Jacob C, Schneiter R. Mature lipid droplets are accessible to ER luminal proteins. J Cell Sci. 2016;129:3803–15.

    Article  CAS  Google Scholar 

  32. Imamura M, Inoguchi T, Ikuyama S, Taniguchi S, Kobayashi K, Nakashima N, et al. ADRP stimulates lipid accumulation and lipid droplet formation in murine fibroblasts. Am J Physiol Endocrinol Metab. 2002;283:E775–83.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This research is supported by the Basic Science Research Program of the Ministry of Education, Science and Technology of Korea (NRF-2016R1A2B4009244).

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Authors and Affiliations

  1. Department of Microbiology and Molecular Biology, Chungnam National University, Daejeon, 34134, Republic of Korea

    Yeonhee Joo, Hyejin Kim & Soojin Lee

  2. Department of Dentistry, Seoul National University, Seoul, 03080, Republic of Korea

    Sungjoong Lee

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Correspondence to Soojin Lee.

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Joo, Y., Kim, H., Lee, S. et al. Neuronal growth regulator 1-deficient mice show increased adiposity and decreased muscle mass. Int J Obes 43, 1769–1782 (2019). https://doi.org/10.1038/s41366-019-0376-2

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