Controlled on-chip stimulation of quantal catecholamine release from chromaffin cells using photolysis of caged Ca2+ on transparent indium-tin-oxide microchip electrodes

被引:36
作者
Chen, Xiaohui [1 ,2 ]
Gao, Yuanfang [3 ]
Hossain, Maruf [3 ]
Gangopadhyay, Shubhra [3 ,4 ]
Gillis, Kevin D. [1 ,2 ,5 ]
机构
[1] Univ Missouri, Dept Biol Engn, Columbia, MO 65211 USA
[2] Univ Missouri, Dalton Cardiovasc Res Ctr, Columbia, MO 65211 USA
[3] Univ Missouri, Dept Elect & Comp Engn, Columbia, MO 65211 USA
[4] Univ Missouri, Dept Med Pharmacol & Physiol, Columbia, MO 65211 USA
[5] Dalton Cardiovasc Res Ctr, Columbia, MO 65211 USA
关键词
D O I
10.1039/b715308m
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Photorelease of caged Ca2+ is a uniquely powerful tool to study the dynamics of Ca2+-triggered exocytosis from individual cells. Using photolithography and other microfabrication techniques, we have developed transparent microchip devices to enable photorelease of caged Ca2+, together with electrochemical detection of quantal catecholamine secretion from individual cells or cell arrays as a step towards developing high-throughput experimental devices. A 100 nm thick transparent indium-tin-oxide (ITO) film was sputter-deposited onto glass coverslips, which were then patterned into 24 cell-sized working electrodes (similar to 20 mu m by 20 mu m). We loaded bovine chromaffin cells with acetoxymethyl (AM) ester derivatives of the Ca2+ cage NP-EGTA and Ca2+ indicator dye fura-4F, then transferred these cells onto the working ITO electrodes for amperometric recordings. Upon flash photorelease of caged Ca2+, a uniform rise of [Ca2+](i) within the target cell leads to quantal release of oxidizable catecholamines measured amperometrically by the underlying ITO electrode. We observed a burst of amperometric spikes upon rapid elevation of [Ca2+](i) and a '' priming '' effect of sub-stimulatory [Ca2+](i) on the response of cells to subsequent [Ca2+](i) elevation, similar to previous reports using different techniques. We conclude that UV photolysis of caged Ca2+ is a suitable stimulation technique for higher-throughput studies of Ca2+-dependent exocytosis on transparent electrochemical microelectrode arrays.
引用
收藏
页码:161 / 169
页数:9
相关论文
共 37 条
[1]   The exocytotic event in chromaffin cells revealed by patch amperometry [J].
Albillos, A ;
Dernick, G ;
Horstmann, H ;
Almers, W ;
deToledo, GA ;
Lindau, M .
NATURE, 1997, 389 (6650) :509-512
[2]   Coupling of electrochemistry and fluorescence microscopy at indium tin oxide microelectrodes for the analysis of single exocytotic events [J].
Amatore, C ;
Arbault, S ;
Chen, Y ;
Crozatier, C ;
Lemaître, F ;
Verchier, Y .
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2006, 45 (24) :4000-4003
[3]   ARTIFICIAL PHOTOSYNTHESIS - SOLAR SPLITTING OF WATER TO HYDROGEN AND OXYGEN [J].
BARD, AJ ;
FOX, MA .
ACCOUNTS OF CHEMICAL RESEARCH, 1995, 28 (03) :141-145
[4]  
BECQUEREL ME, 1839, HEBD SEANCES ACAD SC, V9, P561
[5]  
BITTNER MA, 1992, J BIOL CHEM, V267, P16219
[6]   Amperometric detection of quantal catecholamine secretion from individual cells on micromachined silicon chips [J].
Chen, P ;
Xu, B ;
Tokranova, N ;
Feng, XJ ;
Castracane, J ;
Gillis, KD .
ANALYTICAL CHEMISTRY, 2003, 75 (03) :518-524
[7]   DELAY IN VESICLE FUSION REVEALED BY ELECTROCHEMICAL MONITORING OF SINGLE SECRETORY EVENTS IN ADRENAL CHROMAFFIN CELLS [J].
CHOW, RH ;
VONRUDEN, L ;
NEHER, E .
NATURE, 1992, 356 (6364) :60-63
[8]   In situ temporal detection of dopamine exocytosis from L-dopa-incubated MN9D cells using microelectrode array-integrated biochip [J].
Cui, Hui-Fang ;
Ye, Jian-Shan ;
Chen, Yu ;
Chong, Ser-Choong ;
Liu, Xiao ;
Lim, Tit-Meng ;
Sheu, Fwu-Shan .
SENSORS AND ACTUATORS B-CHEMICAL, 2006, 115 (02) :634-641
[9]   Characteristics of indium tin oxide films deposited by DC and RF magnetron sputtering [J].
Deng, WL ;
Ohgi, T ;
Nejo, H ;
Fujita, D .
JAPANESE JOURNAL OF APPLIED PHYSICS PART 1-REGULAR PAPERS SHORT NOTES & REVIEW PAPERS, 2001, 40 (5A) :3364-3369
[10]  
Di Carlo D, 2006, ANAL CHEM, V78, P7918