Mechanical stimulation of a single cell in an airway epithelial culture initiates an increase in intracellular Ca2+ concentration ([Ca2+]i) that propagates from cell to cell as an intercellular Ca2+ wave. These Ca2+ waves appear to require an increase in intracellular inositol 1,4,5-trisphosphate (IP3) concentration ([IP3]i) in the stimulated cell and are propagated between cells by the diffusion of IP3 through gap junctions. To test the hypothesis that the activation of phospholipase C (PLC) contributes to the elevation of [IP3]i and initiation of an intercellular Ca2+ wave, changes in [Ca2+]i induced by mechanical stimulation were measured by digital fluorescence microscopy in the presence of the PLC inhibitor, aminosteroid U73122. Following exposure to U73122 mechanical stimulation elevated [Ca2+]i of the stimulated cell, but did not initiate the propagation of an intercellular Ca2+ wave. By contrast, in the presence of U73343, a similar aminosteroid that does not inactivate PLC, mechanical stimulation increased the [Ca2+]i of the stimulated cell and initiated an intercellular Ca2+ wave. U73122 also blocked the elevation of [Ca2+]i of airway epithelial cells in response to ATP, a P2-receptor agonist that activates PLC to elevate [IP3]i and [Ca2+]i. In addition, the propagation of intercellular Ca2+ waves was not affected by the ryanodine-receptor agonists, caffeine or ryanodine. The hypotheses that: (1) an elevation of [IP3]i is required to initiate intercellular Ca2+ waves; (2) mechanical stimulation activates PLC; and (3) Ca2+ wave propagation in airway epithelial cells involves Ca2+ release from intracellular stores primarily via IP3 receptors are supported by these results.

REFERENCES

Amundson
J.
,
Clapham
D.
(
1993
).
Calcium waves.
Curr. Opin. Neurobiol
3
,
375
382
Bala
G. A.
,
Thakur
N. R.
,
Bleasdale
J. E.
(
1990
).
Characterization of the major phosphoinositide-specific phospholipase C of human amnion.
Biol. Reprod
43
,
704
711
Bleasdale
J. E.
,
Thakur
N. R.
,
Gremban
R. S.
,
Bundy
G. L.
,
Fitzpatrick
F. A.
,
Smith
R. J.
,
Bunting
S.
(
1990
).
Selective inhibition of receptor-coupled phospholipase C-dependent processes in human platelets and polymorphonuclear neutrophils.
J. Pharmacol. Exp. Ther
255
,
756
768
Boitano
S.
,
Dirksen
E. R.
,
Sanderson
M. J.
(
1992
).
Intercellular propagation of calcium waves mediated by inositol trisphosphate.
Science
258
,
292
295
Boitano.
S.
,
Dirksen
E. R.
,
Sanderson
M. J.
(
1994
).
The role of intercellular Ca2+signalling in epithelial cells.
FASEB J
8
,
664
–.
Boitano
S.
,
Sanderson
M. J.
,
Dirksen
E. R.
(
1994
).
A role for Ca2+-conducting ion channels in mechanically-induced signal transduction of airway epithelial cells.
J. Cell Sci
107
,
3037
3044
Brophy
C. M.
,
Mills
I.
,
Rosales
O.
,
Isales
C.
,
Sumpio
B. E.
(
1993
).
Phospholipase C: A putative mechanotransducer for endothelial cell responses to acute hemodynamic changes.
Biochem. Biophys. Res. Commun
190
,
576
581
Carter
T. D.
,
Chen
X. Y.
,
Carlile
G.
,
Kalapothakis
E.
,
Ogden
D. C.
,
Evans
W. H.
(
1994
).
Cultured porcine aortic endothelial gap junctions: characterization and permeation by second messenger molecules.
J. Physiol
475
,
74
–.
Charles
A. C.
,
Merrill
J. E.
,
Dirksen
E. R.
,
Sanderson
M. J.
(
1991
).
Intercellular signaling in glial cells: calcium waves and oscillations in response to mechanical stimulation and glutamate.
Neuron
6
,
983
992
Charles
A. C.
,
Naus
C. C. G.
,
Zhu
D.
,
Kidder
G. M.
,
Dirksen
E. R.
,
Sanderson
M. J.
(
1992
).
Intercellular calcium signaling via gap junctions in glioma cells.
J. Cell Biol
118
,
195
201
Charles
A. C.
,
Dirksen
E. R.
,
Merrill
J. E.
,
Sanderson
M. J.
(
1993
).
Mechanisms of intercellular calcium signaling in glial cells studied with dantrolene and thapsigargin.
Glia
7
,
134
145
Cornell-Bell
A. H.
,
Finkbeiner
S. M.
,
Cooper
M. S.
,
Smith
S. J.
(
1990
).
Glutamate induces calcium waves in cultured astrocytes: long-range glial signaling.
Science
247
,
470
473
Dani
J. W.
,
Chernjavsky
A.
,
Smith
S. J.
(
1992
).
Neuronal activity triggers calcium waves in hippocampal astrocyte networks.
Neuron
8
,
429
440
Dassouli
A.
,
Sulpice
J. C.
,
Roux
S.
,
Crozatier
B.
(
1993
).
Stretch-induced inositol trisphosphate and tetrakisphosphate production in cardiomyocytes.
J. Mol. Cell. Cardiol
25
,
973
982
Dubyak
G. R.
(
1991
).
Signal transduction by P2-purinergic receptors for extracellular ATP.
Am. J. Respir. Cell Mol. Biol
4
,
295
300
Finkbeiner
S. M.
(
1992
).
Calcium waves in astrocytes- filling in the gaps.
Neuron
8
,
1101
1108
Friel
D. D.
,
Tsien
R. W.
(
1992
).
Phase-dependent contributions from Ca2+entry and Ca2+release to caffeine-induced [Ca2+]ioscillations in bullfrog sympathetic neurons.
Neuron
8
,
1109
1125
Furuya
K.
,
Enomoto
K.
,
Yamagishi
S.
(
1993
).
Spontaneous calcium oscillations and chemically induced calcium responses in mammary epithelial cells.
Eur. J. Physiol
442
,
295
304
Giannini
G.
,
Clementi
E.
,
Ceci
R.
,
Marziali
G.
,
Sorrention
V.
(
1992
).
Expression of a ryanodine receptor-Ca2+channel that is regulated by TGF-.
Science
257
,
91
94
Goldschmidt-Clermont
P. J.
,
Kim
J. W.
,
Machesky
L. M.
,
Rhee
S. G.
,
Pollard
T. D.
(
1991
).
Regulation of phospholipase C-gamma 1 by profilin and tyrosine phosphorylation.
Science
251
,
1231
1233
Hansen
M.
,
Boitano
S.
,
Dirksen
E. R.
,
Sanderson
M. J.
(
1993
).
Intercellular calcium signaling induced by extracellular adenosine-5-triphosphate and mechanical stimulation in airway epithelial cells.
J. Cell Sci
106
,
995
1004
Hawes
B. E.
,
Waters
S. B.
,
Janovick
J. A.
,
Bleasdale
J. E.
,
Conn
P. M.
(
1992
).
Gonadotropin-releasing hormone-stimulated intracellular Ca2+fluctuations and luteinizing hormone release can be uncoupled from inositol phosphate production.
Endocrinology
130
,
3475
3483
Jaffe
L. F.
(
1993
).
Classes and mechanisms of calcium waves.
Cell Calcium
14
,
736
745
Jin
W.
,
Lo
T. M.
,
Loh
H. H.
,
Thayer
S. A.
(
1994
).
U73122 inhibits phospholipase C-dependent calcium mobilization in neuronal cells.
Brain Res
642
,
237
243
Kim
W. T.
,
Rioult
M. G.
,
Cornell-Bell
A. H.
(
1994
).
Glutamate-induced calcium signaling in astrocytes.
Glia
11
,
173
184
Kulik
T. J.
,
Bialecki
R. A.
,
Colucci
W. S.
,
Rothman
A.
,
Glennon
E. T.
,
Underwood
R. H.
(
1991
).
Stretch increases inositol trisphosphate and inositol tetrakisphosphate in cultured pulmonary vascular smooth muscle cells.
Biochem. Biophys. Res. Commun
180
,
982
987
Lansley
A. B.
,
Sanderson
M. J.
,
Dirksen
E. R.
(
1992
).
Control of the beat cycle of respiratory tract cilia by Ca2+and cAMP.
Am. J. Physiol
263
,
232
–.
Liedtke
C. M.
(
1994
).
Activation of PtdIns(4, 5)P2-sensitive phospholipase C in rabbit tracheal epithelial cells.
Am. J. Physiol
266
,
397
–.
Marshall
I. C.
,
Taylor
C. W.
(
1993
).
Regulation of inositol 1, 4, 5-trisphosphate receptors.
J. Exp. Biol
184
,
161
182
Matsumoto
H.
,
Baron
C. B.
,
Coburn
R. F.
(
1995
).
Smooth muscle stretch-activated phospholipase C activity.
Am. J. Physiol
268
,
458
–.
McPherson
P. S.
,
Campbell
K. P.
(
1993
).
The ryanodine receptor/Ca2+release channel.
J. Biol. Chem
268
,
13765
13768
Meissner
G.
(
1986
).
Ryanodine activation and inhibition of the Ca2+release channel of sarcoplasmic reticulum.
J. Biol. Chem
261
,
6300
6306
Parys
J. B.
,
Sernett
S. W.
,
DeLisle
S.
,
Snyder
P. M.
,
Welsh
M. J.
,
Campbell
K. P.
(
1992
).
Isolation, characterization and localization of the inositol 1, 4, 5-trisphosphate receptor protein in Xenopus laevis oocytes.
J. Biol. Chem
267
,
18776
18782
Ridgway
E. B.
,
Gilkey
J. C.
,
Jaffe
L. F.
(
1977
).
Free calcium increases explosively in activating medaka eggs.
Proc. Nat. Acad. Sci. USA
74
,
623
627
Rooney
T. A.
,
Thomas
A. P.
(
1993
).
Intracellular calcium waves generated by Ins(1, 4, 5)P3-dependent mechanisms.
Cell Calcium
14
,
674
690
Sáez
J. C.
,
Conner
J. A.
,
Spray
D. C.
,
Bennett
M. V. L.
(
1989
).
Hepatocyte gap junctions are permeable to the second messengers inositol 1, 4, 5-trisphosphate, and to calcium ions.
Proc. Nat. Acad. Sci. USA
86
,
2708
2712
Salari
H.
,
Bramley
A.
,
Langlands
J.
,
Howard
S.
,
Chan-Yeung
M.
,
Chan
H.
,
Schellenberg
R.
(
1994
).
Effect of phospholipase C inhibitor U-73122 on antigen-induced airway smooth muscle contraction in guinea pigs.
Am. J. Respir. Cell Mol. Biol
9
,
405
410
Sanderson.
M. J.
,
Dirksen
E. R.
(
1985
).
A versatile and quantitative computer-assisted photoelectronic technique used for the analysis of ciliary beat cycles.
Cell Motil
5
,
267
292
Sanderson
M. J.
,
Dirksen
E. R.
(
1986
).
Mechanosensitivity of cultured ciliated cells from the mammalian respiratory tract: implications for the regulation of mucociliary transport.
Proc. Nat. Acad. Sci. USA
83
,
7302
7306
Sanderson
M. J.
,
Dirksen
E. R.
(
1989
).
Mechanosensitive and beta-adrenergic control of the ciliary beat frequency of mammalian respiratory tract cells in culture.
Am. Rev. Respir. Dis
139
,
432
440
Sanderson
M. J.
,
Charles
A. C.
,
Dirksen
E. R.
(
1990
).
Mechanical stimulation and intercellular communication increases intracellular Ca2+in epithelial cells.
Cell Regul
1
,
585
596
Sanderson
M. J.
,
Charles
A. C.
,
Dirksen
E. R.
(
1993
).
Measurement of the temporospatial dynamics of intercellular calcium signaling with digital fluorescence microscopy.
Am. Lab
25
,
29
36
Sanderson
M. J.
,
Charles
A. C.
,
Boitano
S.
,
Dirksen
E. R.
(
1994
).
Mechanisms and function of intercellular calcium signaling.
Mol. Cell. Endocrinol
98
,
173
187
Sanderson
M. J.
(
1995
).
Intercellular calcium waves mediated by inositol trisphosphate. In Calcium, Waves, Gradients and Oscillations.
CIBA Foundation Symposium
188
,
175
194
Smallridge
R. C.
,
Kiang
J. G.
,
Gist
I. D.
,
Fein
H. G.
,
Galloway
R. J.
(
1992
).
U-73122, an aminosteroid phospholipase C antagonist, noncompetitively inhibits thyrotropin-releasing hormone effects in GH3 rat pituitary cells.
Endocrinology
131
,
1883
1888
Smith
R. J.
,
Sam
L. M.
,
Justen
J. M.
,
Bundy
G. L.
,
Bala
G. A.
,
Bleasdale
J. E.
(
1990
).
Receptor-coupled signal transduction in human polymorphonuclear neutrophils: effects of a novel inhibitor of phospholipase C-dependent processes on cell responsiveness.
J. Pharmacol. Exp. Ther
253
,
688
697
Sneyd
J.
,
Charles
A. C.
,
Sanderson
M. J.
(
1994
).
A model for the propagation of intercellular calcium waves.
Am. J. Physiol
266
,
293
–.
Tamaoki
J.
,
Kondo
M.
,
Takizawa
T.
(
1989
).
Effect of cAMP on ciliary function in rabbit tracheal epithelial cells.
J. Appl. Physiol
66
,
1035
1039
Thompson
A. K.
,
Mostafapour
S. P.
,
Denlinger
L. C.
,
Bleasdale
J. E.
,
Fisher
S. K.
(
1991
).
The aminosteroid U-73122 inhibits muscarinic receptor sequestration and phosphoinositide hydrolysis in SK-N-SH neuroblastoma cells. A role for Gp in receptor compartmentation.
J. Biol. Chem
266
,
23856
23862
Warburton
D.
,
Buckley
S.
,
Cosico
L.
(
1989
).
P1and P2purinergic receptor signal transduction in rat type II pneumocytes.
J. Appl. Physiol
66
,
901
905
Yule
D. I.
,
Williams
J. A.
(
1992
).
U73122 inhibits Ca2+oscillations in response to cholecystokinin and carbachol but not to JMV-180 in rat pancreatic acinar cells.
J. Biol. Chem
267
,
13830
13835
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