Abstract
Lipid second messengers play important roles in many cell signaling cascades. Lipid signaling molecules allow for high specificity, rapid transduction, and rapid reversibility of localized stimulation events. Fluorescent sensors capable of detecting individual signaling lipids enable their production and degradation to be followed, revealing the nature and dynamics of signaling pathways. The following sections outline a method for using lipid sensors to monitor the production of signaling lipids on the plasma membrane of C2C12 myotubes in response to insulin signaling.
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References
Balla, T., Bondeva, T., and Varnai, P. (2000) How accurately can we image inositol lipids in living cells? Trends Pharmacol Sci 21, 238–241
Hurley, J. H., and Meyer, T. (2001) Subcellular targeting by membrane lipids. Curr Opin Cell Biol 13, 146–152
Spiegel, S., Foster, D., and Kolesnick, R. (1996) Signal transduction through lipid second messengers. Curr Opin Cell Biol 8, 159–167
Morone, N., Fujiwara, T., Murase, K., Kasai, R. S., Ike, H., Yuasa, S., Usukura, J., and Kusumi, A. (2006) Three-dimensional reconstruction of the membrane skeleton at the plasma membrane interface by electron tomography. J Cell Biol 174, 851–862
Ghosh, S., Strum, J. C., Sciorra, V. A., Daniel, L., and Bell, R. M. (1996) Raf-1 kinase possesses distinct binding domains for phosphatidylserine and phosphatidic acid. Phosphatidic acid regulates the translocation of Raf-1 in 12-O-tetradecanoylphorbol-13-acetate-stimulated Madin-Darby canine kidney cells. J Biol Chem 271, 8472–8480
Oancea, E., Teruel, M. N., Quest, A. F., and Meyer, T. (1998) Green fluorescent protein (GFP)-tagged cysteine-rich domains from protein kinase C as fluorescent indicators for diacylglycerol signaling in living cells. J Cell Biol 140, 485–498
Holz, R. W., Hlubek, M. D., Sorensen, S. D., Fisher, S. K., Balla, T., Ozaki, S., Prestwich, G. D., Stuenkel, E. L., and Bittner, M. A. (2000) A pleckstrin homology domain specific for phosphatidylinositol 4, 5-bisphosphate (PtdIns-4,5-P2) and fused to green fluorescent protein identifies plasma membrane PtdIns-4,5-P2 as being important in exocytosis. J Biol Chem 275, 17878–17885
Huang, P., Altshuller, Y. M., Hou, J. C., Pessin, J. E., and Frohman, M. A. (2005) Insulin-stimulated plasma membrane fusion of Glut4 glucose transporter-containing vesicles is regulated by phospholipase D1. Mol Biol Cell 16, 2614–2623
Zeniou-Meyer, M., Zabari, N., Ashery, U., Chasserot-Golaz, S., Haeberle, A. M., Demais, V., Bailly, Y., Gottfried, I., Nakanishi, H., Neiman, A. M., Du, G., Frohman, M. A., Bader, M. F., and Vitale, N. (2007) Phospholipase D1 production of phosphatidic acid at the plasma membrane promotes exocytosis of large dense-core granules at a late stage. J Biol Chem 282, 21746–21757
Su, W., Yeku, O., Olepu, S., Genna, A., Park, J. S., Ren, H., Du, G., Gelb, M., Morris, A., and Frohman, M. A. (2009) FIPI: A Phospholipase D pharmacological inhibitor that alters cell spreading and inhibits chemotaxis. Mol. Pharm. 75, 437– 466
Nakanishi, H., de los Santos, P., and Neiman, A. M. (2004) Positive and negative regulation of a SNARE protein by control of intracellular localization. Mol Biol Cell 15, 1802–1815
Corrotte, M., Chasserot-Golaz, S., Huang, P., Du, G., Ktistakis, N. T., Frohman, M. A., Vitale, N., Bader, M. F., and Grant, N. J. (2006) Dynamics and function of phospholipase D and phosphatidic acid during phagocytosis. Traffic (Copenhagen, Denmark) 7, 365–377
Zhao, C., Du, G., Skowronek, K., Frohman, M. A., and Bar-Sagi, D. (2007) Phospholipase D2-generated phosphatidic acid couples EGFR stimulation to Ras activation by Sos. Nat Cell Biol 9, 707–712
Gray, A., Van Der Kaay, J., and Downes, C. P. (1999) The pleckstrin homology domains of protein kinase B and GRP1 (general receptor for phosphoinositides-1) are sensitive and selective probes for the cellular detection of phosphatidylinositol 3,4-bisphosphate and/or phosphatidylinositol 3,4,5-trisphosphate in vivo. Biochem J 344 Pt 3, 929–936
Lindsay, Y., McCoull, D., Davidson, L., Leslie, N. R., Fairservice, A., Gray, A., Lucocq, J., and Downes, C. P. (2006) Localization of agonist-sensitive PtdIns(3,4,5) P3 reveals a nuclear pool that is insensitive to PTEN expression. J Cell Sci 119, 5160–5168
Blatner, N. R., Stahelin, R. V., Diraviyam, K., Hawkins, P. T., Hong, W., Murray, D., and Cho, W. (2004) The molecular basis of the differential subcellular localization of FYVE domains. J Biol Chem 279, 53818–53827
Bogan, J. S., McKee, A. E., and Lodish, H. F. (2001) Insulin-responsive compartments containing glut4 in 3t3-l1 and cho cells: regulation by amino acid concentrations. Mol Cell Biol 21, 4785–4806
Shaner, N. C., Campbell, R. E., Steinbach, P. A., Giepmans, B. N., Palmer, A. E., and Tsien, R. Y. (2004) Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat Biotechnol 22, 1567–1572
Shaner, N. C., Steinbach, P. A., and Tsien, R. Y. (2005) A guide to choosing fluorescent proteins. Nature Methods 2, 905–909
Nedachi, T., and Kanzaki, M. (2006) Regulation of glucose transporters by insulin and extracellular glucose in C2C12 myotubes. Am J Physiol Endocrinol Metab 291, E817–828
Gallegos, L. L., Kunkel, M. T., and Newton, A. C. (2006) Targeting protein kinase C activity reporter to discrete intracellular regions reveals spatiotemporal differences in agonist-dependent signaling. J Biol Chem 281, 30947–30956
Sato, M., Ueda, Y., and Umezawa, Y. (2006) Imaging diacylglycerol dynamics at organelle membranes. Nature Methods 3, 797–799
Acknowledgments
This work was supported by NIH award GM071520 to MAF and an NIH NIDDK NRSA award and the Turner Foundation to MO.
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Osisami, M., Huang, H., Frohman, M.A. (2009). Monitoring Insulin-Stimulated Production of Signaling Lipids at the Plasma Membrane. In: Park-Sarge, OK., Curry, T. (eds) Molecular Endocrinology. Methods in Molecular Biology, vol 590. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-378-7_4
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DOI: https://doi.org/10.1007/978-1-60327-378-7_4
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