Change in intracellular calcium ions during shear induced platelet aggregation

K. Kawakami, M. Fukuyama, K. Sakai, I. Itagaki, K. Kawano, M. Handa, Y. Ikeda

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The authors have developed a method to measure intracellular calcium ion concentration ([Ca2+]i) during shear-induced platelet aggregation. A cone and plate viscometer was adapted for continuous recording of both light transmission and fluorescence intensity. Citrated platelet rich plasma was incubated with Indo-1AM at a concentration of 10 μm for 30 min at 37° C, then applied to the albumin density gradient to prepare washed platelets. To Indo-1AM loaded washed platelets, fibrinogen, and von Willebrand factor (vWf) were added, with 1 mM CaCl2. Platelets were then exposed to changing shear stress (6-108 dynes/cm2) for simultaneous measurement of aggregation and [Ca2+]i. [Ca2+]i in resting platelets was estimated as approximately 100 nM. At low shear stress (10-20 dynes/cm2), [Ca2+]i did not change. In contrast, a marked increase in [Ca2+]i was observed concurrent with aggregation at high shear stress (100-108 dynes/cm2). However, no increase was seen in the presence of 1 mM EGTA. The increase was prevented by monoclonal antibodies against GPIb or vWf, which inhibited vWf binding to GPIb. A monoclonal anti-vWf antibody, which inhibited vWf binding to the GPIIb/IIIa complex, did not affect [Ca2+]i increase during high shear induced platelet aggregation. These results suggest that binding of vWf to GPIb may trigger Ca2+ influx.

Original languageEnglish
JournalASAIO Transactions
Volume36
Issue number3
Publication statusPublished - 1990 Jul

Fingerprint

von Willebrand Factor
Platelets
Platelet Aggregation
Calcium
Agglomeration
Ions
Blood Platelets
Shear stress
Platelet-Rich Plasma
Egtazic Acid
Monoclonal antibodies
Viscometers
Fibrinogen
Light transmission
Albumins
Antibodies
Fluorescence
Cones
Monoclonal Antibodies
Light

ASJC Scopus subject areas

  • Biophysics

Cite this

Kawakami, K., Fukuyama, M., Sakai, K., Itagaki, I., Kawano, K., Handa, M., & Ikeda, Y. (1990). Change in intracellular calcium ions during shear induced platelet aggregation. ASAIO Transactions, 36(3).

Change in intracellular calcium ions during shear induced platelet aggregation. / Kawakami, K.; Fukuyama, M.; Sakai, K.; Itagaki, I.; Kawano, K.; Handa, M.; Ikeda, Y.

In: ASAIO Transactions, Vol. 36, No. 3, 07.1990.

Research output: Contribution to journalArticle

Kawakami, K, Fukuyama, M, Sakai, K, Itagaki, I, Kawano, K, Handa, M & Ikeda, Y 1990, 'Change in intracellular calcium ions during shear induced platelet aggregation', ASAIO Transactions, vol. 36, no. 3.
Kawakami K, Fukuyama M, Sakai K, Itagaki I, Kawano K, Handa M et al. Change in intracellular calcium ions during shear induced platelet aggregation. ASAIO Transactions. 1990 Jul;36(3).
Kawakami, K. ; Fukuyama, M. ; Sakai, K. ; Itagaki, I. ; Kawano, K. ; Handa, M. ; Ikeda, Y. / Change in intracellular calcium ions during shear induced platelet aggregation. In: ASAIO Transactions. 1990 ; Vol. 36, No. 3.
@article{8a1883d85fa546029cdb61c16daf7eab,
title = "Change in intracellular calcium ions during shear induced platelet aggregation",
abstract = "The authors have developed a method to measure intracellular calcium ion concentration ([Ca2+]i) during shear-induced platelet aggregation. A cone and plate viscometer was adapted for continuous recording of both light transmission and fluorescence intensity. Citrated platelet rich plasma was incubated with Indo-1AM at a concentration of 10 μm for 30 min at 37° C, then applied to the albumin density gradient to prepare washed platelets. To Indo-1AM loaded washed platelets, fibrinogen, and von Willebrand factor (vWf) were added, with 1 mM CaCl2. Platelets were then exposed to changing shear stress (6-108 dynes/cm2) for simultaneous measurement of aggregation and [Ca2+]i. [Ca2+]i in resting platelets was estimated as approximately 100 nM. At low shear stress (10-20 dynes/cm2), [Ca2+]i did not change. In contrast, a marked increase in [Ca2+]i was observed concurrent with aggregation at high shear stress (100-108 dynes/cm2). However, no increase was seen in the presence of 1 mM EGTA. The increase was prevented by monoclonal antibodies against GPIb or vWf, which inhibited vWf binding to GPIb. A monoclonal anti-vWf antibody, which inhibited vWf binding to the GPIIb/IIIa complex, did not affect [Ca2+]i increase during high shear induced platelet aggregation. These results suggest that binding of vWf to GPIb may trigger Ca2+ influx.",
author = "K. Kawakami and M. Fukuyama and K. Sakai and I. Itagaki and K. Kawano and M. Handa and Y. Ikeda",
year = "1990",
month = "7",
language = "English",
volume = "36",
journal = "ASAIO Journal",
issn = "1058-2916",
publisher = "Lippincott Williams and Wilkins",
number = "3",

}

TY - JOUR

T1 - Change in intracellular calcium ions during shear induced platelet aggregation

AU - Kawakami, K.

AU - Fukuyama, M.

AU - Sakai, K.

AU - Itagaki, I.

AU - Kawano, K.

AU - Handa, M.

AU - Ikeda, Y.

PY - 1990/7

Y1 - 1990/7

N2 - The authors have developed a method to measure intracellular calcium ion concentration ([Ca2+]i) during shear-induced platelet aggregation. A cone and plate viscometer was adapted for continuous recording of both light transmission and fluorescence intensity. Citrated platelet rich plasma was incubated with Indo-1AM at a concentration of 10 μm for 30 min at 37° C, then applied to the albumin density gradient to prepare washed platelets. To Indo-1AM loaded washed platelets, fibrinogen, and von Willebrand factor (vWf) were added, with 1 mM CaCl2. Platelets were then exposed to changing shear stress (6-108 dynes/cm2) for simultaneous measurement of aggregation and [Ca2+]i. [Ca2+]i in resting platelets was estimated as approximately 100 nM. At low shear stress (10-20 dynes/cm2), [Ca2+]i did not change. In contrast, a marked increase in [Ca2+]i was observed concurrent with aggregation at high shear stress (100-108 dynes/cm2). However, no increase was seen in the presence of 1 mM EGTA. The increase was prevented by monoclonal antibodies against GPIb or vWf, which inhibited vWf binding to GPIb. A monoclonal anti-vWf antibody, which inhibited vWf binding to the GPIIb/IIIa complex, did not affect [Ca2+]i increase during high shear induced platelet aggregation. These results suggest that binding of vWf to GPIb may trigger Ca2+ influx.

AB - The authors have developed a method to measure intracellular calcium ion concentration ([Ca2+]i) during shear-induced platelet aggregation. A cone and plate viscometer was adapted for continuous recording of both light transmission and fluorescence intensity. Citrated platelet rich plasma was incubated with Indo-1AM at a concentration of 10 μm for 30 min at 37° C, then applied to the albumin density gradient to prepare washed platelets. To Indo-1AM loaded washed platelets, fibrinogen, and von Willebrand factor (vWf) were added, with 1 mM CaCl2. Platelets were then exposed to changing shear stress (6-108 dynes/cm2) for simultaneous measurement of aggregation and [Ca2+]i. [Ca2+]i in resting platelets was estimated as approximately 100 nM. At low shear stress (10-20 dynes/cm2), [Ca2+]i did not change. In contrast, a marked increase in [Ca2+]i was observed concurrent with aggregation at high shear stress (100-108 dynes/cm2). However, no increase was seen in the presence of 1 mM EGTA. The increase was prevented by monoclonal antibodies against GPIb or vWf, which inhibited vWf binding to GPIb. A monoclonal anti-vWf antibody, which inhibited vWf binding to the GPIIb/IIIa complex, did not affect [Ca2+]i increase during high shear induced platelet aggregation. These results suggest that binding of vWf to GPIb may trigger Ca2+ influx.

UR - http://www.scopus.com/inward/record.url?scp=0025457115&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025457115&partnerID=8YFLogxK

M3 - Article

VL - 36

JO - ASAIO Journal

JF - ASAIO Journal

SN - 1058-2916

IS - 3

ER -