CA-DEPENDENT K-CHANNELS IN SMOOTH-MUSCLE CELLS PERMEABILIZED BY BETA-ESCIN RECORDED USING THE CELL-ATTACHED PATCH-CLAMP TECHNIQUE

被引:21
作者
MURAKI, K [1 ]
IMAIZUMI, Y [1 ]
WATANABE, M [1 ]
机构
[1] NAGOYA CITY UNIV,FAC PHARMACEUT SCI,DEPT CHEM PHARMACOL,3-1 TANABEDORI,MIZUHO KU,NAGOYA,AICHI 467,JAPAN
来源
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY | 1992年 / 420卷 / 5-6期
关键词
CA-DEPENDENT K-CHANNEL; BETA-ESCIN; PERMEABILIZATION; SINGLE SMOOTH MUSCLE CELL; ACETYLCHOLINE; SUBSTANCE-P; INOSITOL 1,4,5-TRISPHOSPHATE;
D O I
10.1007/BF00374620
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
Using the cell-attached patch-clamp technique, the activity of single, Ca-dependent K channels was recorded in single smooth muscle cells permeabilized by beta-escin. The conductance and the relationship between the open probability of the channels and pCa recorded in permeabilized cells were very similar to those obtained in excised inside-out patches. At pCa 7, application of 30-mu-M acetylcholine (ACh) or 0.1-mu-M substance P (SP) together with 1 mM guanosine 5'-trisphosphate to permeabilized cells elicited transient bursts of channel openings similar to those which occur in intact cells. Transient activation was also observed when 2-30-mu-M inositol trisphosphate (IP3) was applied to permeabilized cells. This single channel activity was inhibited by pre-treatment with low-molecular-weight heparin at 50-100-mu/ml. Channel activity at pCa 7.0 was greatly enhanced by 200-mu-M cyclic adenosine monophosphate. These results provide direct evidence that single Ca-dependent K channel activity is regulated by the transmitters ACh and SP, as well as a second messenger, IP3, via the release of intracellular Ca from intracellular sites which are blocked by heparin. This novel approach is valuable in elucidating second messenger mechanisms involved in the regulation of single channel activity by transmitters and autacoids, since permeabilization by beta-escin preserves the entire system of receptor-operated signal transduction and allows intracellular application of second messengers at fixed concentrations.
引用
收藏
页码:461 / 469
页数:9
相关论文
共 57 条
  • [1] AKABARALI H, 1990, CAN J PHYSL PHARM, V68, P1489
  • [2] CALCIUM-ACTIVATED POTASSIUM CHANNELS IN SINGLE SMOOTH-MUSCLE CELLS OF RABBIT JEJUNUM AND GUINEA-PIG MESENTERIC-ARTERY
    BENHAM, CD
    BOLTON, TB
    LANG, RJ
    TAKEWAKI, T
    [J]. JOURNAL OF PHYSIOLOGY-LONDON, 1986, 371 : 45 - 67
  • [3] SPONTANEOUS TRANSIENT OUTWARD CURRENTS IN SINGLE VISCERAL AND VASCULAR SMOOTH-MUSCLE CELLS OF THE RABBIT
    BENHAM, CD
    BOLTON, TB
    [J]. JOURNAL OF PHYSIOLOGY-LONDON, 1986, 381 : 385 - 406
  • [4] LITHIUM AMPLIFIES AGONIST-DEPENDENT PHOSPHATIDYLINOSITOL RESPONSES IN BRAIN AND SALIVARY-GLANDS
    BERRIDGE, MJ
    DOWNES, CP
    HANLEY, MR
    [J]. BIOCHEMICAL JOURNAL, 1982, 206 (03) : 587 - 595
  • [5] CARDIAC-MUSCLE FUNCTION - RESULTS FROM SKINNED FIBER PREPARATIONS
    BEST, PM
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY, 1983, 244 (02): : H167 - H177
  • [6] VARIATIONS OF MEMBRANE CHOLESTEROL ALTER THE KINETICS OF CA-2+-DEPENDENT K+ CHANNELS AND MEMBRANE FLUIDITY IN VASCULAR SMOOTH-MUSCLE CELLS
    BOLOTINA, V
    OMELYANENKO, V
    HEYES, B
    RYAN, U
    BREGESTOVSKI, P
    [J]. PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 1989, 415 (03): : 262 - 268
  • [7] PROPERTIES OF CALCIUM STORES AND TRANSIENT OUTWARD CURRENTS IN SINGLE SMOOTH-MUSCLE CELLS OF RABBIT INTESTINE
    BOLTON, TB
    LIM, SP
    [J]. JOURNAL OF PHYSIOLOGY-LONDON, 1989, 409 : 385 - 401
  • [8] INFLUENCE OF LITHIUM ON 2ND MESSENGER ACCUMULATION IN NG108-15 CELLS
    BRAMI, BA
    LELI, U
    HAUSER, G
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1991, 174 (02) : 606 - 612
  • [9] FATTY-ACID MODIFIES CA-2+-DEPENDENT POTASSIUM CHANNEL ACTIVITY IN SMOOTH-MUSCLE CELLS FROM THE HUMAN AORTA
    BREGESTOVSKI, PD
    BOLOTINA, VM
    SEREBRYAKOV, VN
    [J]. PROCEEDINGS OF THE ROYAL SOCIETY SERIES B-BIOLOGICAL SCIENCES, 1989, 237 (1288): : 259 - 266
  • [10] CHARACTERIZATION OF STEREOSPECIFIC BINDING-SITES FOR INOSITOL 1,4,5-TRISPHOSPHATE IN AIRWAY SMOOTH-MUSCLE
    CHILVERS, ER
    CHALLISS, RAJ
    WILLCOCKS, AL
    POTTER, BVL
    BARNES, PJ
    NAHORSKI, SR
    [J]. BRITISH JOURNAL OF PHARMACOLOGY, 1990, 99 (02) : 297 - 302