Stimulation of the antigen receptor on WEHI-231 B lymphoma cells results in a voltage-independent increase in cytoplasmic calcium

Joshua LaBaer, R. Y. Tsien, K. A. Fahey, A. L. DeFranco

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

WEHI-231, a lymphoma-derived murine B cell line, responded to anti-IgM antibodies by increasing the concentration of free calcium in the cytoplasm from 140 nM to 590 nM within 15 sec. This is very similar to the response observed previously in normal B cells (Pozzan et al., 1982, J. Cell Biol. 94:335). Only antibodies specific for mIgM stimulated this response; control antibodies had no effect. In addition, anti-IgM did not stimulate a response by a mutant with a greatly decreased amount of membrane IgM. The relationship of this increase in cytoplasmic calcium to the plasma membrane potential was examined. Anti-IgM did not cause a rapid depolarization of the cells, suggesting that a voltage-dependent calcium channel was not responsible for the calcium increase. Furthermore, experimental depolarization of WEHI-231 cells did not cause a calcium influx, and the calcium increase caused by anti-IgM was not greatly affected by previous depolarization or by prevention of depolarization. These experiments argue strongly that the increase in cytoplasmic calcium is not mediated by a depolarization-activated calcium channel, such as the one found in cardiac muscle and in some neurons. Indeed, a significant portion of the initial increase in cytoplasmic calcium was due to the release of calcium from internal stores, suggesting the involvement of a soluble mediator. Examination of these internal storage sites in permeabilized cells revealed that inositol 1,4,5-trisphosphate could induce the release of calcium. These results are consistent with the hypothesis that the calcium increase in B cells stimulated by anti-IgM is caused by breakdown of phosphatidylinositol 4,5-bisphosphate, generating diacylglycerol and inositol trisphosphate, with the latter compound mediating calcium mobilization.

Original languageEnglish (US)
Pages (from-to)1836-1844
Number of pages9
JournalJournal of Immunology
Volume137
Issue number6
StatePublished - 1986
Externally publishedYes

Fingerprint

Antigen Receptors
B-Cell Lymphoma
Calcium
B-Lymphocytes
Calcium Channels
Calcium Compounds
Inositol 1,4,5-Trisphosphate
Antibodies
Diglycerides
Inositol
Phosphatidylinositols
Membrane Potentials
Immunoglobulin M
Anti-Idiotypic Antibodies
Lymphoma
Myocardium
Cytoplasm
Cell Membrane
anti-IgM
Neurons

ASJC Scopus subject areas

  • Immunology

Cite this

Stimulation of the antigen receptor on WEHI-231 B lymphoma cells results in a voltage-independent increase in cytoplasmic calcium. / LaBaer, Joshua; Tsien, R. Y.; Fahey, K. A.; DeFranco, A. L.

In: Journal of Immunology, Vol. 137, No. 6, 1986, p. 1836-1844.

Research output: Contribution to journalArticle

@article{1af2815e947d4ae0a8fee073831037ca,
title = "Stimulation of the antigen receptor on WEHI-231 B lymphoma cells results in a voltage-independent increase in cytoplasmic calcium",
abstract = "WEHI-231, a lymphoma-derived murine B cell line, responded to anti-IgM antibodies by increasing the concentration of free calcium in the cytoplasm from 140 nM to 590 nM within 15 sec. This is very similar to the response observed previously in normal B cells (Pozzan et al., 1982, J. Cell Biol. 94:335). Only antibodies specific for mIgM stimulated this response; control antibodies had no effect. In addition, anti-IgM did not stimulate a response by a mutant with a greatly decreased amount of membrane IgM. The relationship of this increase in cytoplasmic calcium to the plasma membrane potential was examined. Anti-IgM did not cause a rapid depolarization of the cells, suggesting that a voltage-dependent calcium channel was not responsible for the calcium increase. Furthermore, experimental depolarization of WEHI-231 cells did not cause a calcium influx, and the calcium increase caused by anti-IgM was not greatly affected by previous depolarization or by prevention of depolarization. These experiments argue strongly that the increase in cytoplasmic calcium is not mediated by a depolarization-activated calcium channel, such as the one found in cardiac muscle and in some neurons. Indeed, a significant portion of the initial increase in cytoplasmic calcium was due to the release of calcium from internal stores, suggesting the involvement of a soluble mediator. Examination of these internal storage sites in permeabilized cells revealed that inositol 1,4,5-trisphosphate could induce the release of calcium. These results are consistent with the hypothesis that the calcium increase in B cells stimulated by anti-IgM is caused by breakdown of phosphatidylinositol 4,5-bisphosphate, generating diacylglycerol and inositol trisphosphate, with the latter compound mediating calcium mobilization.",
author = "Joshua LaBaer and Tsien, {R. Y.} and Fahey, {K. A.} and DeFranco, {A. L.}",
year = "1986",
language = "English (US)",
volume = "137",
pages = "1836--1844",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "6",

}

TY - JOUR

T1 - Stimulation of the antigen receptor on WEHI-231 B lymphoma cells results in a voltage-independent increase in cytoplasmic calcium

AU - LaBaer, Joshua

AU - Tsien, R. Y.

AU - Fahey, K. A.

AU - DeFranco, A. L.

PY - 1986

Y1 - 1986

N2 - WEHI-231, a lymphoma-derived murine B cell line, responded to anti-IgM antibodies by increasing the concentration of free calcium in the cytoplasm from 140 nM to 590 nM within 15 sec. This is very similar to the response observed previously in normal B cells (Pozzan et al., 1982, J. Cell Biol. 94:335). Only antibodies specific for mIgM stimulated this response; control antibodies had no effect. In addition, anti-IgM did not stimulate a response by a mutant with a greatly decreased amount of membrane IgM. The relationship of this increase in cytoplasmic calcium to the plasma membrane potential was examined. Anti-IgM did not cause a rapid depolarization of the cells, suggesting that a voltage-dependent calcium channel was not responsible for the calcium increase. Furthermore, experimental depolarization of WEHI-231 cells did not cause a calcium influx, and the calcium increase caused by anti-IgM was not greatly affected by previous depolarization or by prevention of depolarization. These experiments argue strongly that the increase in cytoplasmic calcium is not mediated by a depolarization-activated calcium channel, such as the one found in cardiac muscle and in some neurons. Indeed, a significant portion of the initial increase in cytoplasmic calcium was due to the release of calcium from internal stores, suggesting the involvement of a soluble mediator. Examination of these internal storage sites in permeabilized cells revealed that inositol 1,4,5-trisphosphate could induce the release of calcium. These results are consistent with the hypothesis that the calcium increase in B cells stimulated by anti-IgM is caused by breakdown of phosphatidylinositol 4,5-bisphosphate, generating diacylglycerol and inositol trisphosphate, with the latter compound mediating calcium mobilization.

AB - WEHI-231, a lymphoma-derived murine B cell line, responded to anti-IgM antibodies by increasing the concentration of free calcium in the cytoplasm from 140 nM to 590 nM within 15 sec. This is very similar to the response observed previously in normal B cells (Pozzan et al., 1982, J. Cell Biol. 94:335). Only antibodies specific for mIgM stimulated this response; control antibodies had no effect. In addition, anti-IgM did not stimulate a response by a mutant with a greatly decreased amount of membrane IgM. The relationship of this increase in cytoplasmic calcium to the plasma membrane potential was examined. Anti-IgM did not cause a rapid depolarization of the cells, suggesting that a voltage-dependent calcium channel was not responsible for the calcium increase. Furthermore, experimental depolarization of WEHI-231 cells did not cause a calcium influx, and the calcium increase caused by anti-IgM was not greatly affected by previous depolarization or by prevention of depolarization. These experiments argue strongly that the increase in cytoplasmic calcium is not mediated by a depolarization-activated calcium channel, such as the one found in cardiac muscle and in some neurons. Indeed, a significant portion of the initial increase in cytoplasmic calcium was due to the release of calcium from internal stores, suggesting the involvement of a soluble mediator. Examination of these internal storage sites in permeabilized cells revealed that inositol 1,4,5-trisphosphate could induce the release of calcium. These results are consistent with the hypothesis that the calcium increase in B cells stimulated by anti-IgM is caused by breakdown of phosphatidylinositol 4,5-bisphosphate, generating diacylglycerol and inositol trisphosphate, with the latter compound mediating calcium mobilization.

UR - http://www.scopus.com/inward/record.url?scp=0022508183&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022508183&partnerID=8YFLogxK

M3 - Article

VL - 137

SP - 1836

EP - 1844

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 6

ER -