title: Isolation of sensory neurons of Aplysia californica for patch clamp recordings of glutamatergic currents.
abstract: The marine gastropod mollusk Aplysia californica has a venerable history as a model of nervous system function, with particular significance in studies of learning and memory. The typical preparations for such studies are ones in which the sensory and motoneurons are left intact in a minimally dissected animal, or a technically elaborate neuronal co-culture of individual sensory and motoneurons. Less common is the isolated neuronal preparation in which small clusters of nominally homogeneous neurons are dissociated into single cells in short term culture. Such isolated cells are useful for the biophysical characterization of ion currents using patch clamp techniques, and targeted modulation of these conductances. A protocol for preparing such cultures is described. The protocol takes advantage of the easily identifiable glutamatergic sensory neurons of the pleural and buccal ganglia, and describes their dissociation and minimal maintenance in culture for several days without serum.
authors.affiliation: Division of Marine Biology and Fisheries, Rosenstiel School of Marine and Atmospheric Sciences, University of Miami, USA. lfieber@rsmas.miami.edu
authors.forename: Lynne A
authors.initials: LA
authors.lastname: Fieber
authors.forename: Stephen L
authors.initials: SL
authors.lastname: Carlson
authors.forename: Andrew T
authors.initials: AT
authors.lastname: Kempsell
authors.forename: Justin B
authors.initials: JB
authors.lastname: Greer
authors.forename: Michael C
authors.initials: MC
authors.lastname: Schmale
mesh_terms: D000818:Animals; D001048:Aplysia / Q000166:cytology* / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods*; D017952:Ganglia, Invertebrate / Q000166:cytology; D018698:Glutamic Acid / Q000502:physiology; D018408:Patch-Clamp Techniques / Q000379:methods*; D011984:Sensory Receptor Cells / Q000166:cytology* / Q000502:physiology
chemical_list: D018698:Glutamic Acid
references.citation: J Neurosci. 1996 Dec 1;16(23):7583-98
references.pmid: 8922415
references.citation: J Neurophysiol. 2011 Oct;106(4):1629-36
references.pmid: 21753031
references.citation: J Vis Exp. 2009 Jun 08;(28):
references.pmid: 19506547
references.citation: Cell. 2006 Dec 29;127(7):1453-67
references.pmid: 17190607
references.citation: J Neurophysiol. 2009 Dec;102(6):3753-65
references.pmid: 19828725
references.citation: Proc Natl Acad Sci U S A. 1998 Sep 1;95(18):10938-43
references.pmid: 9724808
references.citation: Science. 1983 Jan 28;219(4583):400-5
references.pmid: 6294833
references.citation: Science. 2001 Nov 2;294(5544):1030-8
references.pmid: 11691980
references.citation: Brain Behav. 2012 Jul;2(4):391-401
references.pmid: 22950043
references.citation: Science. 1970 Mar 27;167(3926):1743-5
references.pmid: 5416542
references.citation: J Neurosci. 1983 Dec;3(12):2403-13
references.pmid: 6655493
references.citation: J Neurosci. 2004 Jul 28;24(30):6833-41
references.pmid: 15282289
references.citation: Cell Mol Neurobiol. 1989 Jun;9(2):233-45
references.pmid: 2743380
references.citation: J Neurophysiol. 1983 Dec;50(6):1522-42
references.pmid: 6663341
references.citation: J Exp Biol. 1998 Mar;201(Pt 5):745-54
references.pmid: 9542153
references.citation: J Neurosci. 1996 Jun 1;16(11):3661-71
references.pmid: 8642410
references.citation: J Neurophysiol. 1978 Mar;41(2):418-31
references.pmid: 650275
references.citation: J Neurophysiol. 1998 Nov;80(5):2514-20
references.pmid: 9819259
references.citation: Neurobiol Learn Mem. 2009 Sep;92(2):147-54
references.pmid: 19332142
references.citation: J Neurophysiol. 2003 Aug;90(2):586-98
references.pmid: 12724359
references.citation: Brain Res. 2010 Jul 9;1343:28-36
references.pmid: 20452331