title: Single cell amperometry reveals glycocalyx hinders the release of neurotransmitters during exocytosis.
abstract: The diffusional hindrance of the glycocalyx along the cell surface on exocytotic peaks, observed with single cell amperometry, was investigated. Partial digestion of the glycocalyx with neuraminidase led to the observation of faster peaks, as shown by varied peak parameters. This result indicates that diffusion of small molecules in the partially digested glycocalyx is 2.2 faster than in the intact glycocalyx. Similarly, neutralization of the negative charges present in the cell microenvironment led to faster peak kinetics. The analysis of the vesicular efflux indicates that the diffusion coefficient of dopamine at the cell surface is at most 45% of the diffusion coefficient in free solution. This study shows that the glycocalyx plays an important role in the diffusion kinetics of processes along the cell surface, including exocytotic events.
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Raphaël
authors.initials: R
authors.lastname: Trouillon
authors.forename: Andrew G
authors.initials: AG
authors.lastname: Ewing
mesh_terms: D000818:Animals; D005089:Exocytosis; D019276:Glycocalyx / Q000737:chemistry / Q000378:metabolism*; D009439:Neuraminidase / Q000737:chemistry / Q000378:metabolism*; D018377:Neurotransmitter Agents / Q000737:chemistry / Q000378:metabolism*; D016716:PC12 Cells; D051381:Rats; D059010:Single-Cell Analysis / Q000379:methods*; D014407:Tumor Cells, Cultured
chemical_list: D018377:Neurotransmitter Agents; D009439:Neuraminidase
references.citation: J Neurosci. 2000 Jul 15;20(14):5276-82
references.pmid: 10884311
references.citation: Am J Physiol Heart Circ Physiol. 2004 Nov;287(5):H2287-94
references.pmid: 15256377
references.citation: Brain Res. 2007 Jun 2;1151:46-54
references.pmid: 17408597
references.citation: J Cell Sci. 1980 Apr;42:81-116
references.pmid: 7400245
references.citation: Chemphyschem. 2010 Jan 18;11(1):159-74
references.pmid: 19937905
references.citation: Mol Biol Cell. 2000 Jul;11(7):2213-9
references.pmid: 10888663
references.citation: Nature. 1983 May 5-11;303(5912):35-40
references.pmid: 6843658
references.citation: Chemphyschem. 2007 Mar 12;8(4):578-85
references.pmid: 17243189
references.citation: Biophys J. 1999 Jan;76(1 Pt 1):509-16
references.pmid: 9876164
references.citation: Nat Rev Neurosci. 2003 Jun;4(6):456-68
references.pmid: 12778118
references.citation: Annu Rev Nutr. 2009;29:177-222
references.pmid: 19575597
references.citation: Anal Chem. 2010 Feb 1;82(3):1020-6
references.pmid: 20039639
references.citation: Nat Methods. 2005 Sep;2(9):651-8
references.pmid: 16118635
references.citation: J Appl Physiol (1985). 1987 Nov;63(5):1987-92
references.pmid: 3693232
references.citation: J Am Soc Nephrol. 2007 Nov;18(11):2885-93
references.pmid: 17942961
references.citation: Am J Physiol. 1994 Apr;266(4 Pt 2):H1300-3
references.pmid: 8184907
references.citation: Chemphyschem. 2007 Dec 3;8(17):2471-7
references.pmid: 17966970
references.citation: Science. 1993 Feb 12;259(5097):963-5
references.pmid: 8438154
references.citation: Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10754-8
references.pmid: 1961743
references.citation: Sci Rep. 2012;2:907
references.pmid: 23205269
references.citation: Nature. 1990 Aug 2;346(6283):425-34
references.pmid: 1974032
references.citation: J Biol Chem. 1982 Oct 25;257(20):11966-70
references.pmid: 7118922
references.citation: Biophys J. 2011 Feb 16;100(4):968-77
references.pmid: 21320441
references.citation: Biochim Biophys Acta. 2007 Jun;1773(6):728-35
references.pmid: 17467077
references.citation: Rev Neurosci. 2000;11(2-3):159-212
references.pmid: 10718152
references.citation: J Neurosci. 2008 Mar 26;28(13):3350-8
references.pmid: 18367602
references.citation: Circ Res. 2002 Nov 15;91(10):877-87
references.pmid: 12433832
references.citation: Nature. 1992 Mar 5;356(6364):60-3
references.pmid: 1538782
references.citation: Biochim Biophys Acta. 1989 Sep 25;1005(2):109-17
references.pmid: 2775765
references.citation: Geriatr Gerontol Int. 2010 Jul;10 Suppl 1:S32-40
references.pmid: 20590840
references.citation: Cell. 2006 Sep 8;126(5):855-67
references.pmid: 16959566
references.citation: ACS Chem Neurosci. 2010 Mar 17;1(3):234-245
references.pmid: 20368748
references.citation: J Neurosci. 2004 Jan 14;24(2):303-9
references.pmid: 14724228
references.citation: Acta Neurochir (Wien). 2004 Dec;146(12):1337-40; discussion 1340
references.pmid: 15340874
references.citation: J Am Chem Soc. 1997 Jan 29;119(4):681-90
references.pmid: 16526129
references.citation: PLoS One. 2011;6(11):e27820
references.pmid: 22125627
references.citation: Biophys J. 2008 May 15;94(10):4114-22
references.pmid: 18234815
references.citation: Nat Chem. 2011 Jul 22;3(8):582-9
references.pmid: 21778976
references.citation: J Immunogenet. 1990 Aug-Oct;17(4-5):219-25
references.pmid: 2093725
references.citation: Phys Chem Chem Phys. 2010 Sep 14;12(34):10065-72
references.pmid: 20625576
references.citation: Biophys J. 1992 Oct;63(4):1118-32
references.pmid: 1420930
references.citation: J Gen Physiol. 1966 Mar;49(4):717-25
references.pmid: 5221375
references.citation: J Cell Sci. 1976 Jun;21(1):129-59
references.pmid: 932106
references.citation: J Cell Biol. 1979 May;81(2):275-300
references.pmid: 38256
references.citation: Eur Biophys J. 1998;27(1):9-17
references.pmid: 9463886
references.citation: Chembiochem. 2006 Dec;7(12):1998-2003
references.pmid: 17086558
references.citation: J Cell Physiol. 2007 Dec;213(3):565-73
references.pmid: 17680633
references.citation: Anal Chem. 2011 Jun 15;83(12):4369-92
references.pmid: 21500835
references.citation: Proc Natl Acad Sci U S A. 2005 Sep 27;102(39):13879-84
references.pmid: 16172395
references.citation: Anal Chem. 2013 Jan 15;85(2):522-42
references.pmid: 23151043
references.citation: Nature. 1993 Jun 10;363(6429):554-8
references.pmid: 8505984
references.citation: Biophys J. 1995 Jan;68(1):383-90
references.pmid: 7711264
references.citation: Physiol Rev. 2004 Apr;84(2):649-98
references.pmid: 15044685
references.citation: J Biol Chem. 1994 Jun 24;269(25):17215-20
references.pmid: 8006030
references.citation: Neurobiol Aging. 2002 Mar-Apr;23(2):287-93
references.pmid: 11804714
references.citation: Biochim Biophys Acta. 1964 Nov 1;83:363-6
references.pmid: 14236715
references.citation: Biophys J. 1996 Feb;70(2):1061-8
references.pmid: 8789125
references.citation: Curr Opin Cell Biol. 2003 Oct;15(5):621-32
references.pmid: 14519398
references.citation: Biophys J. 1996 Aug;71(2):1131-9
references.pmid: 8842250