Abstract
Understanding how signaling molecules are released from cells is essential for furthering our knowledge of the basic biological mechanisms controlling many significant biological pathways. These molecules, including neurotransmitters, hormones, growth factors, and peptides, are released from cells via a process called exocytosis. Our laboratory has utilized a noninvasive method of measuring the release of oxidizable molecules from cells, known as carbon-fiber amperometry. In this chapter we will describe how we undertake such measurements, how the resulting data is analyzed, and what the outcomes mean in terms of physiology. We provide examples of our work measuring catecholamine release in single chromaffin cells as well as serotonin release from intact sections of colon.
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References
Burgoyne RD, Morgan A (2003) Secretory granule exocytosis. Physiol Rev 83:581–632
Sudhof TC (2004) The synaptic vesicle cycle. Annu Rev Neurosci 27:509–547
Kissinger PT, Hart JB, Adams RN (1973) Voltammetry in brain tissue—a new neurophysiological measurement. Brain Res 55:209–213
Chow R, Rüden L (2009) Electrochemical detection of secretion from single cells. In: Sakmann B, Neher E (eds) Single-channel recording. Springer, New York, pp 245–275
Wightman RM, Jankowski JA, Kennedy RT et al (1991) Temporally resolved catecholamine spikes correspond to single vesicle release from individual chromaffin cells. Proc Natl Acad Sci U S A 88:10754–10758
Stamford JA (1986) In vivo voltammetry: some methodological considerations. J Neuro-sci Methods 17:1–29
Leszczyszyn DJ, Jankowski JA, Viveros OH, Diliberto EJ, Near JA, Wightman RM (1990) Nicotinic receptor-mediated catecholamine secretion from individual chromaffin cells. Chemical evidence for exocytosis. J Biol Chem 265:14736–14737
Keating DJ, Dubach D, Zanin MP et al (2008) Dscr1/rcan1 regulates vesicle exocytosis and fusion pore kinetics: implications for Down syndrome and Alzheimer’s disease. Hum Mol Genet 17:1020–1030
Zanin MP, Phillips L, Mackenzie KD, Keating DJ (2011) Aging differentially affects multiple aspects of vesicle fusion kinetics. PLoS One 6:e27820
Spencer NJ, Nicholas SJ, Robinson L et al (2011) Mechanisms underlying distension-evoked peristalsis in guinea pig distal colon: is there a role for enterochromaffin cells? Am J Physiol 301:G519–G527
Keating DJ, Spencer NJ (2010) Release of 5-hydroxytryptamine from the mucosa is not required for the generation or propagation of colonic migrating motor complexes. Gastroenterology 138:659–670, e652
Chow RH, von Ruden L, Neher E (1992) Delay in vesicle fusion revealed by electrochemical monitoring of single secretory events in adrenal chromaffin cells. Nature 356:60–63
Zhou Z, Misler S, Chow RH (1996) Rapid fluctuations in transmitter release from single vesicles in bovine adrenal chromaffin cells. Biophys J 70:1543–1552
Douglas WW (1968) Stimulus-secretion coupling: the concept and clues from chromaffin and other cells. Br J Pharmacol 34:451–474
Burgoyne RD (1991) Control of exocytosis in adrenal chromaffin cells. Biochim Biophys Acta 1071:174–202
DomÃnguez N, RodrÃguez M, Machado JD, Borges R (2012) Preparation and culture of adrenal chromaffin cells # T neurotrophic factors. Methods Mol Biol 846:223–234
Moser T, Neher E (1997) Rapid exocytosis in single chromaffin cells recorded from mouse adrenal slices. J Neurosci 17:2314–2323
Colliver TL, Hess EJ, Ewing AG (2001) Amperometric analysis of exocytosis at chromaffin cells from genetically distinct mice. Neurosci Methods 105:95–103
Erspamer V (1954) Pharmacology of indole-alkylamines. Pharmacol Rev 6:425–487
Thompson M, Fleming KA, Evans DJ, Fundele R, Surani MA, Wright NA (1990) Gastric endocrine cells share a clonal origin with other gut cell lineages. Development 110:477–481
Racke K, Schworer H (1991) Regulation of serotonin release from the intestinal mucosa. Pharmacol Res 23:13–25
Racke K, Reimann A, Schworer H, Kilbinger H (1996) Regulation of 5-ht release from enterochromaffin cells. Behav Brain Res 73:83–87
Minami M, Tamakai H, Ogawa T et al (1995) Chemical modulation of 5-ht3 and 5-ht4 receptors affects the release of 5-hydroxytryptamine from the ferret and rat intestine. Res Commun Mol Pathol Pharmacol 89:131–142
Hirafuji M, Ogawa T, Kato K et al (2001) Noradrenaline stimulates 5-hydroxytryptamine release from mouse ileal tissues via alpha(2)-adrenoceptors. Eur J Pharmacol 432:149–152
Bertrand PP (2006) Real-time measurement of serotonin release and motility in guinea pig ileum. J Physiol 577:689–704
Tanaka T, Mizumoto A, Mochiki E, Haga N, Suzuki H, Itoh Z (2004) Relationship between intraduodenal 5-hydroxytryptamine release and interdigestive contractions in dogs. J Smooth Muscle Res 40:75–84
Bertrand PP, Bertrand RL (2010) Serotonin release and uptake in the gastrointestinal tract. Auton Neurosci 153:47–57
Bertrand PP (2004) Real-time detection of serotonin release from enterochromaffin cells of the guinea-pig ileum. Neurogastroenterol Motil 16:511–514
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Duffield, M.D., Raghupathi, R., Keating, D.J. (2014). Carbon-Fiber Amperometry in the Study of Exocytosis. In: Thorn, P. (eds) Exocytosis Methods. Neuromethods, vol 83. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-676-4_3
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DOI: https://doi.org/10.1007/978-1-62703-676-4_3
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Publisher Name: Humana Press, Totowa, NJ
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