Imaging Lipid Metabolism at the Golgi Complex

  • Serena CapassoEmail author
  • Giovanni D’Angelo
Part of the Methods in Molecular Biology book series (MIMB, volume 1949)


The development of fluorescence-based molecular imaging has revolutionized cell biology allowing the visualization of specific biomolecules at the microscopic and, more recently, at the nanoscopic scale while in their relevant biological contexts. Nonetheless, despite the imaging toolkit for biologists interested in exploring the subcellular localization and dynamics of proteins and nucleic acids has expanded exponentially in the last decades, the means to visualize and track lipids in cells did not develop to the same extent until recently. Here we described some basic fluorescence-based techniques that can be used in standard cell biology laboratories to visualize subcellular pools of specific lipids and to evaluate their regional metabolism. Specifically, here we focus on the imaging-based analysis of phosphoinositide and sphingolipid metabolism at the Golgi complex.

Key words

Immunofluorescence Sphingolipids Phosphatidylinositol-4-Phosphate Diacylglycerol Golgi complex 


  1. 1.
    Hanada K, Kumagai K, Yasuda S et al (2003) Molecular machinery for non-vesicular trafficking of ceramide. Nature 426:803–809CrossRefGoogle Scholar
  2. 2.
    Hannun YA, Obeid LM (2008) Principles of bioactive lipid signalling: lessons from sphingolipids. Nat Rev Mol Cell Biol 9:139–150CrossRefGoogle Scholar
  3. 3.
    Hu W, Ross J, Geng T et al (2011) Differential regulation of dihydroceramide desaturase by palmitate versus monounsaturated fatty acids: implications for insulin resistance. J Biol Chem 286:16596–16605CrossRefGoogle Scholar
  4. 4.
    Breslow DK, Weissman JS (2010) Membranes in balance: mechanisms of sphingolipid homeostasis. Mol Cell 40:267–279CrossRefGoogle Scholar
  5. 5.
    Capasso S, Sticco L, Rizzo R et al (2017) Sphingolipid metabolic flow controls phosphoinositide turnover at the trans-Golgi network. EMBO J 36:1736–1754CrossRefGoogle Scholar
  6. 6.
    D’Angelo G, Polishchuk E, Di Tullio G et al (2007) Glycosphingolipid synthesis requires FAPP2 transfer of glucosylceramide. Nature 449:62–67CrossRefGoogle Scholar
  7. 7.
    D’Angelo G, Uemura T, Chuang CC et al (2013) Vesicular and non-vesicular transport feed distinct glycosylation pathways in the Golgi. Nature 501:116–120CrossRefGoogle Scholar
  8. 8.
    Tolias KF, Cantley LC (1999) Pathways for phosphoinositide synthesis. Chem Phys Lipids 98:69–77CrossRefGoogle Scholar
  9. 9.
    Ebrahimzadeh Z, Mukherjee A, Richard D (2018) A map of the subcellular distribution of phosphoinositides in the erythrocytic cycle of the malaria parasite Plasmodium falciparum. Int J Parasitol 48:13–25CrossRefGoogle Scholar
  10. 10.
    Mesmin B, Bigay J, Moser von Filseck J et al (2013) A four-step cycle driven by PI(4)P hydrolysis directs sterol/PI(4)P exchange by the ER-Golgi tether OSBP. Cell 155:830–843CrossRefGoogle Scholar
  11. 11.
    Hsuan J, Cockcroft S (2001) The PITP family of phosphatidylinositol transfer proteins. Genome Biol 2:Reviews3011.1–Reviews3011.8CrossRefGoogle Scholar
  12. 12.
    Lemmon MA (2010) Chapter 136—Pleckstrin homology (PH) domains. In: Bradshaw RA, Dennis EA (eds) Handbook of cell signaling, 2nd edn. Academic Press, San Diego, pp 1093–1101CrossRefGoogle Scholar
  13. 13.
    Cheong FY, Sharma V, Blagoveshchenskaya A et al (2010) Spatial regulation of Golgi phosphatidylinositol-4-phosphate is required for enzyme localization and glycosylation fidelity. Traffic 11:1180–1190CrossRefGoogle Scholar
  14. 14.
    Hammond GR, Machner MP, Balla T (2014) A novel probe for phosphatidylinositol 4-phosphate reveals multiple pools beyond the Golgi. J Cell Biol 205:113–126CrossRefGoogle Scholar
  15. 15.
    Hammond GR, Schiavo G, Irvine RF (2009) Immunocytochemical techniques reveal multiple, distinct cellular pools of PtdIns4P and PtdIns(4,5)P(2). Biochem J 422:23–35CrossRefGoogle Scholar
  16. 16.
    Wuttke A, Idevall-Hagren O, Tengholm A (2010) Imaging phosphoinositide dynamics in living cells. Methods Mol Biol 645:219–235CrossRefGoogle Scholar
  17. 17.
    Levine TP, Munro S (1998) The pleckstrin homology domain of oxysterol-binding protein recognises a determinant specific to Golgi membranes. Curr Biol 8:729–739CrossRefGoogle Scholar
  18. 18.
    Baron CL, Malhotra V (2002) Role of diacylglycerol in PKD recruitment to the TGN and protein transport to the plasma membrane. Science 295:325–328CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of Protein Biochemistry, National Council ResearchNaplesItaly

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