Biology; Biophysics; Biotechnology; Chemistry; Nanotechnology; Optics and Photonics
A long-term goal of my laboratory is to understand how the exo-endocytic cycle is spatially regulated during cellular morphogenesis. Two major focuses are the spatial-temporal regulation of membrane traffic in adipocytes in response to insulin signaling and in fibroblasts during cell migration. To elucidated mechanisms that regulate membrane trafficking, my laboratory uses advanced imaging and analysis coupled with RNAi knockdown of key regulator machinery (e.g. exocyst complex). Innovative live cell TIRFM and spinning-disk confocal imaging at the 'CINEMA' imaging center are used to visualize directly, at the single-vesicle level, where and how membrane traffic is delivered and regulated. As impaired vesicle exocytosis is associated with type 2 diabetes and abnormal cell motility is associated with metastasis and cancer, the knowledge gained with this research will be highly significant in understanding the pathologies of these diseases.
Specialized Terms: Cell Biology; Exocytosis; Endocytosis; Membrane Traffic; Tethering; Microscopy; Live Cell Imaging; TIRFM; Single molecule
Extensive Research Description
Cellular Imaging and Analysis of Polarized Membrane Traffic. A major goal of my laboratory is to develop and apply new and state-of-the-art multidimensional optical methods to better understand the basic mechanisms of polarized membrane trafficking and spatial-temporal control of endo-exocytosis.
One important challenge facing modern biology is to understand how individual biochemical reactions are integrated in space and time. Increasingly, new vital probes and optical methods has begun to provide unique mechanistic insight into how molecules, vesicles, organelles and whole cells are (re)organized in response to internal and external cues. This is especially relevant for the dynamic process of membrane traffic and the cytoskeleton in cell polarity - key areas of our interest. Insight into how cells both establish and lose polarity are also essential for understanding disease processes such as metastasis. In particular we are applying the optical methods of Total Internal Reflections Fluorescence Microscopy (TIRFM) and 4D (3D + time) multicolor spinning-disk confocal imaging to directly address, at the single-vesicle level, where and how polarized membrane traffic is delivered. TIRFM imaging (also called evanescent wave microscopy) can selectively illuminate an extremely thin optical section (
Specifically, using advance optical methods our lab is exploring the following related topics:
1) organization and coordination of exocytosis and cytoskeleton in polarized cells and
2) coupling of exo- and endocytosis and molecular mechanisms that regulate this process. For instance, TIRFM imaging has lead to a number of novel observations including imaging of constitutive exocytosis (and the surprising presence of exocytic ‘hot-spots’ for fusion on the cell surface) and nanometer targeting of microtubule plus ends to the cell surface and focal adhesions. To facilitate these and other studies multicolor TIRFM instruments, a 4D spinning disk confocal and electrophysiology instrumentation has been recently implemented here as part of “The CINEMA Lab” ("Cinema Imaging Using New Microscopy Approaches"), with support from Ludwig Institute for Cancer Research (LICR), various grants, Yale and the private sector. We are also collaborating with other groups at Yale (Dr. Jim Duncan’s group, Dept. of Biomedical Engineering) and overseas (Elena Diaz, Spain) to develop novel software to detect, analyze and make computational cellular models of these processes.
phosphoinositide switch controls the maturation and signaling properties of APPL endosomes.
Zoncu R, Perera RM, Balkin DM, Pirruccello M, Toomre D, De Camilli P. A phosphoinositide switch controls the maturation and signaling properties of APPL endosomes. Cell. 2009 Mar 20;136(6):1110-21. PubMed PMID: 19303853; PubMed Central PMCID: PMC2705806.
Two synaptojanin 1 isoforms are recruited to clathrin-coated pits at different stages.
Perera RM, Zoncu R, Lucast L, De Camilli P, Toomre D. Two synaptojanin 1 isoforms are recruited to clathrin-coated pits at different stages. Proc Natl Acad Sci U S A. 2006 Dec 19;103(51):19332-7. Epub 2006 Dec 8.
Spatio-temporal analysis of constitutive exocytosis in epithelial cells.
Sebastian R, Diaz ME, Ayala G, Letinic K, Moncho-Bogani J, Toomre D. Spatio-temporal analysis of constitutive exocytosis in epithelial cells. IEEE/ACM Trans Comput Biol Bioinform. 2006 Jan-Mar;3(1):17-32.
Both daughter cells traffic and exocytose membrane at the cleavage furrow during mammalian cytokinesis.
Goss JW, Toomre DK. Both daughter cells traffic and exocytose membrane at the cleavage furrow during mammalian cytokinesis. J Cell Biol. 2008 Jun 30;181(7):1047-54. Epub 2008 Jun 23. PubMed PMID: 18573914; PubMed Central PMCID: PMC2442215.
Fusion of constitutive membrane traffic with the cell surface observed by evanescent wave microscopy.
Toomre D, Steyer JA, Keller P, Almers W, Simons K. Fusion of constitutive membrane traffic with the cell surface observed by evanescent wave microscopy. J Cell Biol. 2000 Apr 3;149(1):33-40. PubMed PMID: 10747085; PubMed Central PMCID: PMC2175107.
Lipid rafts and signal transduction.
Simons K, Toomre D. Lipid rafts and signal transduction. Nat Rev Mol Cell Biol. 2000 Oct;1(1):31-9. Review. Erratum in: Nat Rev Mol Cell Biol 2001 Mar;2(3):216. PubMed PMID: 11413487.
Full List of PubMed Publications
- Schepartz A, Thompson AD, Omar MH, Rivera-Molina F, Xi Z, Koleske AJ, Toomre DK: Long-Term Live-Cell STED Nanoscopy of Primary and Cultured Cells with the Plasma Membrane HIDE Probe DiI-SiR. Angew Chem Int Ed Engl. 2017 Jul 5; 2017 Jul 5. PMID: 28679029
- Xu Y, Toomre DK, Bogan JS, Hao M: Excess cholesterol inhibits glucose-stimulated fusion pore dynamics in insulin exocytosis. J Cell Mol Med. 2017 May 25; 2017 May 25. PMID: 28544529
- Hoang D, Broer N, Sosa JA, Abitbol N, Yao X, Li F, Rivera-Molina F, Toomre DK, Roman SA, Sue G, Kim S, Li AY, Callender GG, Simpson C, Narayan D: Leptin Is Produced by Parathyroid Glands and Stimulates Parathyroid Hormone Secretion. Ann Surg. 2016 Sep 8; 2016 Sep 8. PMID: 27611607
- Deng Y, Rivera-Molina FE, Toomre DK, Burd CG: Sphingomyelin is sorted at the trans Golgi network into a distinct class of secretory vesicle. Proc Natl Acad Sci U S A. 2016 Jun 14; 2016 May 31. PMID: 27247384
- Bottanelli F, Kromann EB, Allgeyer ES, Erdmann RS, Wood Baguley S, Sirinakis G, Schepartz A, Baddeley D, Toomre DK, Rothman JE, Bewersdorf J: Two-colour live-cell nanoscale imaging of intracellular targets. Nat Commun. 2016 Mar 4; 2016 Mar 4. PMID: 26940217
- Idevall-Hagren O, Dickson EJ, Hille B, Toomre DK, De Camilli P: Optogenetic control of phosphoinositide metabolism. Proc Natl Acad Sci U S A. 2012 Aug 28; 2012 Jul 30. PMID: 22847441
- Arasaki K, Toomre DK, Roy CR: The Legionella pneumophila effector DrrA is sufficient to stimulate SNARE-dependent membrane fusion. Cell Host Microbe. 2012 Jan 19. PMID: 22264512
- Xu Y, Melia TJ, Toomre DK: Using light to see and control membrane traffic. Curr Opin Chem Biol. 2011 Dec; 2011 Nov 10. PMID: 22079055
- Xu Y, Rubin BR, Orme CM, Karpikov A, Yu C, Bogan JS, Toomre DK: Dual-mode of insulin action controls GLUT4 vesicle exocytosis. J Cell Biol. 2011 May 16; 2011 May 9. PMID: 21555461
- Liang L, Shen H, De Camilli P, Toomre DK, Duncan JS: An expectation maximization based method for subcellular particle tracking using multi-angle TIRF microscopy. Med Image Comput Comput Assist Interv. 2011. PMID: 22003671
- Goss JW, Toomre DK: Both daughter cells traffic and exocytose membrane at the cleavage furrow during mammalian cytokinesis. J Cell Biol. 2008 Jun 30; 2008 Jun 23. PMID: 18573914
- Iwakiri Y, Satoh A, Chatterjee S, Toomre DK, Chalouni CM, Fulton D, Groszmann RJ, Shah VH, Sessa WC: Nitric oxide synthase generates nitric oxide locally to regulate compartmentalized protein S-nitrosylation and protein trafficking. Proc Natl Acad Sci U S A. 2006 Dec 26; 2006 Dec 14. PMID: 17170139
- Murata T, Delprato A, Ingmundson A, Toomre DK, Lambright DG, Roy CR: The Legionella pneumophila effector protein DrrA is a Rab1 guanine nucleotide-exchange factor. Nat Cell Biol. 2006 Sep; 2006 Aug 13. PMID: 16906144