title: Matching native electrical stimulation by graded chemical stimulation in isolated mouse adrenal chromaffin cells.
abstract: Adrenal chromaffin cells release multiple transmitters in response to sympathetic stimulation. Modest cell firing, matching sympathetic tone, releases small freely soluble catecholamines. Elevated electrical firing rates matching input under sympathetic stress results in release of catecholamines as well as semi-soluble vaso- and neuro-active peptides packaged within the dense core of the secretory granule. This activity-dependent differential transmitter release has been shown to rely on a mechanistic shift in the mode of exocytosis through the regulated dilation of the secretory fusion pore between granule and cell surface membranes. However, biochemical description of the mechanism regulating fusion pore dilation remains elusive. In the experimental setting, electrical stimulation designed to mimic sympathetic input, is achieved through single-cell voltage-clamp. While precise, this approach is incompatible with biochemical and proteomic analysis, both of which require large sample sizes. We address this limitation in the current study. We describe a bulk chemical stimulation paradigm calibrated to match defined electrical activity. We utilize calcium and single-cell amperometric measurements to match extracellular potassium concentrations to physiological electrical stimulation under sympathetic tone as well as acute stress conditions. This approach provides larger samples of uniformly stimulated cells for determining molecular players in activity-dependent differential transmitter release from adrenal chromaffin cells.
authors: Department of Physiology and Biophysics, School of Medicine, 2109 Adelbert Road, Case Western Reserve University, Cleveland, OH 44106-4970, USA. Tiberiu.Fulop@Case.Edu
authors: Tiberiu
authors: T
authors: Fulop
authors: Corey
authors: C
authors: Smith
mesh_terms: D000200:Action Potentials / Q000187:drug effects / Q000528:radiation effects; D000311:Adrenal Glands / Q000166:cytology*; D000704:Analysis of Variance; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002395:Catecholamines / Q000378:metabolism; D019439:Chromaffin Cells / Q000187:drug effects* / Q000528:radiation effects*; D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D004558:Electric Stimulation / Q000379:methods*; D004563:Electrochemistry / Q000379:methods; D051379:Mice; D008810:Mice, Inbred C57BL; D011188:Potassium / Q000494:pharmacology; D013268:Stimulation, Chemical
chemical_list: D002395:Catecholamines; D011188:Potassium; D002118:Calcium
references: J Physiol. 2003 Dec 15;553(Pt 3):707-17
references: 14500763
references: J Physiol. 2004 Oct 15;560(Pt 2):413-28
references: 15297569
references: J Physiol. 1976 Dec;263(3):417-39
references: 1018274
references: J Physiol. 1980 Oct;307:199-216
references: 7205664
references: Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10754-8
references: 1961743
references: Nature. 1992 Mar 5;356(6364):60-3
references: 1538782
references: Nature. 1993 Jun 10;363(6429):554-8
references: 8505984
references: J Anat. 1993 Oct;183 ( Pt 2):237-52
references: 8300414
references: Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12765-9
references: 7809118
references: Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8328-32
references: 7667289
references: Nature. 1997 Oct 2;389(6650):509-12
references: 9333242
references: Int Rev Cytol. 1998;181:213-320
references: 9522458
references: Neuron. 1998 Jun;20(6):1243-53
references: 9655511
references: EMBO J. 2000 Jul 17;19(14):3586-96
references: 10899113
references: J Neurosci Methods. 2000 Nov 30;103(2):151-6
references: 11084207
references: Neuron. 2001 Sep 13;31(5):819-30
references: 11567619
references: Proc Natl Acad Sci U S A. 2002 Apr 30;99(9):6358-63
references: 11959911
references: J Physiol. 2002 May 1;540(Pt 3):921-39
references: 11986380
references: Proc Natl Acad Sci U S A. 2002 May 14;99(10):7124-9
references: 11997474
references: Ann N Y Acad Sci. 2002 Oct;971:647-54
references: 12438200
references: Proc Natl Acad Sci U S A. 2003 Mar 4;100(5):2171-3
references: 12606723
references: Pflugers Arch. 2003 Feb;445(5):540-6
references: 12634923
references: Biochim Biophys Acta. 2003 Aug 18;1641(2-3):167-73
references: 12914957
references: Phys Med Biol. 2003 Oct 21;48(20):3277-93
references: 14620058
references: J Neurosci. 1999 Jan 15;19(2):589-98
references: 9880579
references: Nat Neurosci. 1998 Jul;1(3):192-200
references: 10195143
references: Neuron. 1999 Jul;23(3):607-15
references: 10433271
references: J Neurosci. 2005 Aug 10;25(32):7324-32
references: 16093382
references: Biochem J. 2006 Oct 1;399(1):111-9
references: 16784416