Center for Matrix Biology - Vanderbilt University and Medical Center

David W. Piston, Ph.D.
Professor in the Department of Molecular Physiology & Biophysics

	Office Address:	702 Light Hall
	Mailing Address:	702 Light Hall Nashville, TN 37232-0615
	Lab address:	747 Light Hall
	Phone:	(615) 322-7030
		email \| website

Secondary appointment in the Physics and Astronomy department.

Research Summary

The research in our lab focuses on molecular events involved in glucose metabolism and insulin secretion in beta cells within intact functioning pancreatic islets. We have developed unique, state-of-the-art fluorescence imaging methods to assay the response at various points along the glucose transduction pathway. The quantitative microscopy is combined with standard biochemical and molecular biological techniques to obtain unique information that bridges the gap between the known details of the glucose transduction pathway in individual beta cells and the overall glucose response of a whole islet. As glucose is metabolized by the beta cell, there is a rise in NAD(P)H that can be monitored by its autofluorescence. Imaging of the temporal and spatial pattern of NAD(P)H has revealed that individual beta cells within the islet form a much more uniform population than was expected from studies of dissociated cells. Glucokinase dominates the rate of glucose signal transduction by beta cells, and we are investigating its allosteric regulation by protein-protein interactions with nitric oxide synthase. This interaction has not been found in vitro, but is considerable in vivo. We are also developing methods to correlate metabolic behavior to beta cell function by imaging cytoplasmic calcium activity, membrane potential, intracellular pH, and secretion of fluorescent insulin. Recently proposed models suggest that gap junctions and/or K+ channels play a role in insulin secretion from whole islets. Our new methods allow us to test these models' predictions, and thus permit determination of the important cell-cell communication mechanisms in the islet.

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Associated initiative(s):

Tissue Regeneration

Career opportunities:
None currently available

Selected Publications

» Kremers, GJ, Hazelwood, KL, Murphy, CS, Davidson, MW, Piston, DW Photoconversion in orange and red fluorescent proteins Nat Methods 2009. 6:355-8

» Easley, CJ, Rocheleau, JV, Head, WS, Piston, DW Quantitative measurement of zinc secretion from pancreatic islets with high temporal resolution using droplet-based microfluidics Anal Chem 2009. 81:9086-95

» Hilliard, FA, Steele, DS, Laver, D, Yang, Z, Le Marchand, SJ, Chopra, N, Piston, DW, Huke, S, Knollmann, BC Flecainide inhibits arrhythmogenic Ca(2+) waves by open state block of ryanodine receptor Ca(2+) release channels and reduction of Ca(2+) spark mass J Mol Cell Cardiol 2009. :

» Gunawardana, SC, Benninger, RK, Piston, DW Subcutaneous transplantation of embryonic pancreas for correction of type 1 diabetes Am J Physiol Endocrinol Metab 2009. 296:E323-32

» Easley, CJ, Benninger, RK, Shaver, JH, Steven Head, W, Piston, DW Rapid and inexpensive fabrication of polymeric microfluidic devices via toner transfer masking Lab Chip 2009. 9:1119-27

» de Jonge, N, Peckys, DB, Kremers, GJ, Piston, DW Electron microscopy of whole cells in liquid with nanometer resolution Proc Natl Acad Sci U S A 2009. 106:2159-64

» Pound, LD, Sarkar, SA, Benninger, RK, Wang, Y, Suwanichkul, A, Shadoan, MK, Printz, RL, Oeser, JK, Lee, CE, Piston, DW, McGuinness, OP, Hutton, JC, Powell, DR, O''Brien, RM Deletion of the mouse Slc30a8 gene encoding zinc transporter-8 results in impaired insulin secretion Biochem J 2009. 421:371-6

» Zavalin, A, Hachey, DL, Sundaramoorthy, M, Banerjee, S, Morgan, S, Feldman, L, Tolk, N, Piston, DW Kinetics of a collagen-like polypeptide fragmentation after mid-IR free-electron laser ablation Biophys J 2008. 95:1371-81

» Gunawardana, SC, Head, WS, Piston, DW Dimethyl amiloride improves glucose homeostasis in mouse models of type 2 diabetes Am J Physiol Endocrinol Metab 2008. 294:E1097-108

» Benninger, RK, Hao, M, Piston, DW Multi-photon excitation imaging of dynamic processes in living cells and tissues Rev Physiol Biochem Pharmacol 2008. 160:71-92

» Benninger, RK, Ashby, WJ, Ring, EA, Piston, DW Single-photon-counting detector for increased sensitivity in two-photon laser scanning microscopy Opt Lett 2008. 33:2895-7

» Nyman, LR, Wells, KS, Head, WS, McCaughey, M, Ford, E, Brissova, M, Piston, DW, Powers, AC Real-time, multidimensional in vivo imaging used to investigate blood flow in mouse pancreatic islets J Clin Invest 2008. 118:3790-7

» Benninger, RK, Zhang, M, Head, WS, Satin, LS, Piston, DW Gap junction coupling and calcium waves in the pancreatic islet Biophys J 2008. 95:5048-61

» Martin, MD, Fingleton, B, Lynch, CC, Wells, S, McIntyre, JO, Piston, DW, Matrisian, LM Establishment and quantitative imaging of a 3D lung organotypic model of mammary tumor outgrowth Clin Exp Metastasis 2008. 25:877-85

» Cricenti, A, Generosi, R, Luce, M, Perfetti, P, Sanghera, JS, Aggarwal, ID, Tolk, NH, Vobornik, D, Margaritondo, G, Piston, DW, Manni, V, Grimaldi, S, Lisi, A, Rieti, S Low-frequency electromagnetic field effects on functional groups in human skin keratinocytes cells revealed by IR-SNOM J Microsc 2008. 229:551-4

» Generosi, J, Margaritondo, G, Sanghera, JS, Aggarwal, ID, Tolk, NH, Piston, DW, Castellano, AC, Cricenti, A Infrared scanning near-field optical microscopy investigates order and clusters in model membranes J Microsc 2008. 229:259-63

» Mao, S, Benninger, RK, Yan, Y, Petchprayoon, C, Jackson, D, Easley, CJ, Piston, DW, Marriott, G Optical lock-in detection of FRET using synthetic and genetically encoded optical switches Biophys J 2008. 94:4515-24

» Zhang, Q, Wang, SY, Fleuriel, C, Leprince, D, Rocheleau, JV, Piston, DW, Goodman, RH Metabolic regulation of SIRT1 transcription via a HIC1:CtBP corepressor complex Proc Natl Acad Sci U S A 2007. 104:829-33

» Gruen, ML, Hao, M, Piston, DW, Hasty, AH Leptin requires canonical migratory signaling pathways for induction of monocyte and macrophage chemotaxis Am J Physiol Cell Physiol 2007. 293:C1481-8

» Hao, M, Head, WS, Gunawardana, SC, Hasty, AH, Piston, DW Direct effect of cholesterol on insulin secretion: a novel mechanism for pancreatic {beta}-cell dysfunction Diabetes 2007. 56:2328-2338

» Li, C, Hao, M, Cao, Z, Ding, W, Graves-Deal, R, Hu, J, Piston, DW, Coffey, RJ Naked2 Acts as a Cargo Recognition and Targeting Protein to Ensure Proper Delivery and Fusion of TGF-{alpha}-containing Exocytic Vesicles at the Lower Lateral Membrane of Polarized MDCK Cells Mol Biol Cell 2007. 18:3081-3093

» Malo, GD, Pouwels, LJ, Wang, M, Weichsel, A, Montfort, WR, Rizzo, MA, Piston, DW, Wachter, RM X-ray Structure of Cerulean GFP: A Tryptophan-Based Chromophore Useful for Fluorescence Lifetime Imaging(,) Biochemistry 2007. 46:9865-9873

» Piston, DW, Kremers, GJ Fluorescent protein FRET: the good, the bad and the ugly Trends Biochem Sci 2007. 32:407-14

» Remedi, MS, Rocheleau, JV, Tong, A, Patton, BL, McDaniel, ML, Piston, DW, Koster, JC, Nichols, CG Hyperinsulinism in mice with heterozygous loss of K(ATP) channels Diabetologia 2006. 49:2368-78

» Zhang, Q, Wang, SY, Nottke, AC, Rocheleau, JV, Piston, DW, Goodman, RH Redox sensor CtBP mediates hypoxia-induced tumor cell migration Proc Natl Acad Sci U S A 2006. 103:9029-33

» Hara, M, Dizon, RF, Glick, BS, Lee, CS, Kaestner, KH, Piston, DW, Bindokas, VP Imaging pancreatic beta-cells in the intact pancreas Am J Physiol Endocrinol Metab 2006. 290:E1041-7

» Rocheleau, JV, Remedi, MS, Granada, B, Head, WS, Koster, JC, Nichols, CG, Piston, DW Critical Role of Gap Junction Coupled K(ATP)Channel Activity for Regulated Insulin Secretion PLoS Biol 2006. 4:e26

» Gunawardana, SC, Rocheleau, JV, Head, WS, Piston, DW Mechanisms of time-dependent potentiation of insulin release: involvement of nitric oxide synthase Diabetes 2006. 55:1029-33

» Rizzo, MA, Springer, G, Segawa, K, Zipfel, WR, Piston, DW Optimization of pairings and detection conditions for measurement of FRET between cyan and yellow fluorescent proteins Microsc Microanal 2006. 12:238-54

» Gunawardana, SC, Head, WS, Piston, DW Amiloride derivatives enhance insulin release in pancreatic islets from diabetic mice BMC Endocr Disord 2005. 5:9

» Gunawardana, SC, Hara, M, Bell, GI, Head, WS, Magnuson, MA, Piston, DW Imaging Beta cell development in real-time using pancreatic explants from mice with green fluorescent protein-labeled pancreatic Beta cells In Vitro Cell Dev Biol Anim 2005. 41:7-11

» Park, SY, Wang, X, Chen, Z, Powers, AC, Magnuson, MA, Head, WS, Piston, DW, Bell, GI Optical imaging of pancreatic beta cells in living mice expressing a mouse insulin I promoter-firefly luciferase transgene Genesis 2005. 43:80-6

» Schwartz, JW, Novarino, G, Piston, DW, Defelice, LJ Substrate binding stoichiometry and kinetics of the norepinephrine transporter J Biol Chem 2005. :

» Rizzo, MA, Piston, DW High-contrast imaging of fluorescent protein FRET by fluorescence polarization microscopy Biophys J 2005. 88:L14-6

» Hao, M, Li, X, Rizzo, MA, Rocheleau, JV, Dawant, BM, Piston, DW Regulation of two insulin granule populations within the reserve pool by distinct calcium sources J Cell Sci 2005. 118:5873-84

» Rocheleau, Jonathan V, Walker, Glenn M, Head, W Steven, McGuinness, Owen P, Piston, David W Microfluidic glucose stimulation reveals limited coordination of intracellular Ca2+ activity oscillations in pancreatic islets Proc Natl Acad Sci U S A 2004. 101:12899-903

» Rizzo, Mark A, Springer, Gerald H, Granada, Butch, Piston, David W An improved cyan fluorescent protein variant useful for FRET Nat Biotechnol 2004. 22:445-9

» Rocheleau, Jonathan V, Head, W Steven, Piston, David W Quantitative NAD(P)H/flavoprotein autofluorescence imaging reveals metabolic mechanisms of pancreatic islet pyruvate response J Biol Chem 2004. 279:31780-7

» Cricenti, A, Generosi, R, Luce, M, Perfetti, P, Margaritondo, G, Talley, D, Sanghera, JS, Aggarwal, ID, Tolk, NH, Congiu-Castellano, A, Rizzo, MA, Piston, DW Chemically resolved imaging of biological cells and thin films by infrared scanning near-field optical microscopy Biophys J 2003. 85:2705-10

» Rocheleau, JV, Piston, DW Two-photon excitation microscopy for the study of living cells and tissues Curr Protoc Cell Biol 2003. Chapter 4:Unit 4.11

» Rocheleau, Jonathan V, Edidin, Michael, Piston, David W Intrasequence GFP in Class I MHC Molecules, a Rigid Probe for Fluorescence Anisotropy Measurements of the Membrane Environment Biophys J 2003. 84:4078-86

» Rizzo, Mark A, Piston, David W Regulation of beta cell glucokinase by S-nitrosylation and association with nitric oxide synthase J Cell Biol 2003. 161:243-8

» Zhang, Qinghong, Piston, David W, Goodman, Richard H Regulation of corepressor function by nuclear NADH Science 2002. 295:1895-7

» Patterson, G H, Knobel, S M, Arkhammar, P, Thastrup, O, Piston, D W Separation of the glucose-stimulated cytoplasmic and mitochondrial NAD(P)H responses in pancreatic islet beta cells Proc Natl Acad Sci U S A 2000. 97:5203-7

» Xu, Y, Piston, D W, Johnson, C H A bioluminescence resonance energy transfer (BRET) system: application to interacting circadian clock proteins Proc Natl Acad Sci U S A 1999. 96:151-6

» Patterson, G H, Knobel, S M, Sharif, W D, Kain, S R, Piston, D W Use of the green fluorescent protein and its mutants in quantitative fluorescence microscopy Biophys J 1997. 73:2782-90

» Bennett, B D, Jetton, T L, Ying, G, Magnuson, M A, Piston, D W Quantitative subcellular imaging of glucose metabolism within intact pancreatic islets J Biol Chem 1996. 271:3647-51

» Niswender, K D, Blackman, S M, Rohde, L, Magnuson, M A, Piston, D W Quantitative imaging of green fluorescent protein in cultured cells: comparison of microscopic techniques, use in fusion proteins and detection limits J Microsc 1995. 180 ( Pt 2):109-16