(tolbutamide), Ca 2 + ( [ Ca 2 + ] i ). (MCh) [ Ca 2 + ] i, [ Ca 2 + ] i Tsinghua Tongfang Optical Disc Co., Ltd. All rights reserved.

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1 mental Botany, 1987, 38 : Zhang J, Daveis W J Abscisic acid produced in dehydrating roots may be the plant to measure the water status of the soil Plant Cell and Environment, 1989, 12 : Davies W J, Zhang J Root signals and the regulation of growth and development of plants in drying soil Ann Rev Plant Physiology and Plant Mol Bio, 1991, 42 : Jackson G E, Irvine J, Grace J, et al Abscisic acid concentration and fluxes in draughted conifer saplings Plant Cell Environ, 1995, 18 : Liang J, Zhang J, Wong M H Can stomatal closure caused by xylem ABA explain the inhibition of leaf photosynthesis under soil dry2 ing? Photosynthesis Research, 1997, 51(2) : ,, : :, Chen S, Wang S, Altman A, et al Genotype- specific differences in drought tolerance of poplar in relation to abscisic acid In : Bach I Environmental and Soil Issue in Poplar and Willow Cultivation and Utilization 20 th Session of the International Poplar Commission Budapest : Hungary, Zhang J, Davies W J Does ABA in the xylem control the rate of leaf growth in soil-dried Maize and sunflower plants?j Experimental Botany, 1990, 41 (230) : Quarrie S A, Whitford P N, Appleford N E J, et al A monoclonal antibody to (s)-abscisic acid : its characterization and use in a ra2 dioimmunoassy for measuring abscisic acid in crud extracts of cereal and lupin leaves Planta, 1988, 173 : Zeevaart J A D, Creelman R A Metabolism and physiology of abscisic acid Ann Rev Plant Physiol Plant Mol Biol, 1988, 39 : Johansson I, Larsson C, Ek B, et al The major integral proteins of spinach leaf plasme membranes are putative aquaporins and are phosphorylated in response to Ca 2 + and apoplastic water potential The Plant Cell, 1996, 8 : ( , ) 3 (, ;, ;, E2mail : edu cn ), Ca 2 +, KCl (tolbutamide), Ca 2 + ( [ Ca 2 + ] i ), 5 mmol/ L (caffeine) (MCh) [ Ca 2 + ] i, IP 3 ryanodine [ Ca 2 + ] i 30 5 s, IP 3 fura22, [1 ] Tsinghua Tongfang Optical Disc Co, Ltd All rights reserved

2 ,, [ Ca 2 + [2,3 ] i ],, [ Ca 2 + ] i Ca 2 + Ca 2 + [4 7 ], [ Ca 2 + ] i, ; caffeine MCh,,, Ca ( ) V,,HEPES,, Sigma ; RPMI21640,Dispase,,BSA Gibco ; KRBB ( Krebs2Ringer bicarbonate buffer) : 129 mmol/ L NaCl, 4 7 mmol/ L KCl, 1 2 mmol/ L KH 2 P0 4, 2 5 mmol/ L CaCl 2, 1 2 mmol/ L MgSO 4, 5 0 mmol/ L NaHCO 3, 10 mmol/ L HEPES, 3 mmol/ L Glucose, BSA 0 1 %, ph = 7 4 KRBB 74 mmol/ L NaCl 60 mmol/ L KCl, 60 mmol/ L KRBB CaCl mmol/ L EGTA, Ca 2 + KRBB 10 % ( ) RPMI ( ) [ 8 ] Dispase : g Wistar,, 2 mg/ ml V, 37, 250 r/ min 30 min KRBB 7 8, mmol/ L EGTA 8 min, Ca 2 +, Mg 2 + KRBB 0 3 mg/ ml Dispase 10 min KRBB, 37 5 %, m ( ) [ Ca 2 + ] i 3 8 d KRBB, Fura22/ AM 3 mol/ L, min Zeiss Axiovert 100 TILL fura22 F 345 F 380, F 345 / F 380 [ Ca 2 + [7,9,10 ] i ] 1 Hz (28 2) ( ) PP283 (Narishige, ) 5 m, KRBB Ca 2 + KRBB, MX21 (Nar2 ishige, ) 10 m,,, [11 ] 2 2 1, 20 mmol/ L, Tsinghua Tongfang Optical Disc Co, Ltd All rights reserved

3 (70 %) F 345 / F 380 ( [ Ca 2 + ] i ) ( 1) : (1) ; (2) ( ATP ATP K(ATP) ) ; (3) Ca (a) ; (b) [ Ca 2 + ] i, 1 (a) ( n = 5), (phase 0) s ; (phase 1), 10, (phase 2), 10, 1 (b) ( n = 7), 0 50 s ( n = 4) ;, ( n = 3) ; Ca 2 + [ Ca 2 + ] i ( n = 2, ) 2 2 ATP ATP ( K(ATP) ),, [ Ca 2 + ] i [12 ] tolbutamide K(ATP) [11,12 ] tolbutamide, [ Ca 2 + ] i, 2 (a) ( n = 7) 5 10 s, 50 % 70 % [ Ca 2 + ] i ( 50 % [11 ] [3, 70 % ] ),, Ca 2 + [ Ca 2 + ] i Ca 2 +,,, [ Ca 2 + ] i ;, [ Ca 2 + ] i, 2 (b) ( n = 4) Tsinghua Tongfang Optical Disc Co, Ltd All rights reserved

4 , [ Ca 2 + ] i 2 (a) tolbutamide [ Ca 2 + ] i ; (b) [ Ca 2 + ] i 2 3 [ Ca 2 + ] i ryanodine, ryanodine 1) ryanodine, KRBB 5 mmol/ L 3 (a), s [ Ca 2 + ] i ( n = 9), 3 (b),, 11 7 s( 13 5 s, n = 4) ; 33 2 s( 28 7 s, n = 4) ryanodine 10 s caffeine 5 mmol/ L caffeine, ryanodine Ca 2 + [13 ], 30 s Ca 2 + ( Ca 2 + Na + / Ca 2 + ) Ca 2 + [13,14 ] [15 (MCh), IP 3 ] Ca 2 + KRBB 5 mmol/ L MCh, 5 s [ Ca 2 + ] i, 4 (a) ( n = 7), 4 (b),, 3 8 s( 4 4 s, n = 4) ; 52 3 s ( 58 9 s, n = 4) ryanodine, 1),,,, 1999 ( ) Tsinghua Tongfang Optical Disc Co, Ltd All rights reserved

5 (a) [ Ca 2 + ] i ; (b) 4 (a) [ Ca 2 + ] i ; (b) Ca 2 + caffeine [ Ca 2 + ] i, Tsinghua Tongfang Optical Disc Co, Ltd All rights reserved

6 ,IP 3 [ Ca 2 + ] i, IP 3,[Ca 2 + ] i,ip 3, 3 fura22 Ca 2 + Ca 2 + : (1) ; (2) Ca 2 +, ryanodine IP 3 ; (3) MCh, ; (4) IP 3 ryanodine 2, Roe [16 ] :,, ( ),,, MCh, IP 3 ryanodine,, ( : ), ( : ), The Li Foundation Heritage Prize 1 Polonsky K S The 2cell in diabetes : from molecular genetics to clinical research DIABETES,1995,44 : Wollheim C B, Sharp G W Regulation of Insulin Release by calcium Physiol Rev, 1981,61 : Ashcroft F M, Proks P, Smith P A, et al Stimulus2secretion coupling in pancreatic cells Journal of Cellular Biochemistry,1994, 55 (suppl) : ,,,, 1997, 13 (3) : ,,,, 1998, 43 (15) : , PLA2801,1998, 43(5) : Zhou Z, Neher E Mobile and Immobile Ca buffers in chromaffin cells J Physiol (London), 1993, 469 : ,,,1998,5 : Grynkiewicy G, Poenie M, Tsien R Y A new generation of Ca 2 + indicators J Bio Chem, 1985, 260 : ,, ( ), 1995, Zhou Z, Misler S Amperonetric detection of quantal secretion from patch2champed rat pancreatic cells J Biol Chem, 1996, 271 : Tsinghua Tongfang Optical Disc Co, Ltd All rights reserved 2063

7 Misler S, Falke L C, Gillis K, et al A metabolite2regulated potassium channel in rat pancreatic 2cells Proc Natl Acad Sci USA, 1986,83 : Bers D Excitation2contraction coupling and cardiac contractile force Dordrecht : Kluwer Academic Press, Herrington J, Park YB, Babcock D F, et al Dominant role of mitochondrial in clearance of large Ca loads from rat adrenal chromaffin cells Neuron,1996, 16 : ,,, SH2SY5Y,1995, 12 : Roe M W, Lancaster M E, Mertz R J, et al Voltage2dependent intracellular calcium release from mouse islets stimulated by glucose J Biol Chem, 1993,268 : ( , ) 3 Bt 3 (, ;, ;, ;, E2mail : ptt js cn) 3 Bt,, R19 3 Bt, R19 Bt, R19 3, Bt ( Bacillus thuringiensis) Bt Bt, [1 ] 1992 Bt [2 ],,, Bt, [3,4 ], ( ) 1 Bt R Tsinghua Tongfang Optical Disc Co, Ltd All rights reserved