Cell shrinkage as a signal to apoptosis in NIH 3T3 fibroblasts.

Martin B Friis; Christel R Friborg; Linda Schneider; Maj-Britt Nielsen; Ian H Lambert; Søren T Christensen; Else K Hoffmann

doi:10.1113/jphysiol.2005.087130

Cell shrinkage as a signal to apoptosis in NIH 3T3 fibroblasts.

Martin B Friis, Christel R Friborg, Linda Schneider, Maj-Britt Nielsen, Ian H Lambert, Søren T Christensen, Else K Hoffmann

Cell Biology and Physiology

124 Citations (Scopus)

Abstract

Cell shrinkage is a hallmark of the apoptotic mode of programmed cell death, but it is as yet unclear whether a reduction in cell volume is a primary activation signal of apoptosis. Here we studied the effect of an acute elevation of osmolarity (NaCl or sucrose additions, final osmolarity 687 mosmol l(-1)) on NIH 3T3 fibroblasts to identify components involved in the signal transduction from shrinkage to apoptosis. After 1.5 h the activity of caspase-3 started to increase followed after 3 h by the appearance of many apoptotic-like bodies. The caspase-3 activity increase was greatly enhanced in cells expressing a constitutively active G protein, Rac (RacV12A3 cell), indicating that Rac acts upstream to caspase-3 activation. The stress-activated protein kinase, p38, was significantly activated by phosphorylation within 30 min after induction of osmotic shrinkage, the phosphorylation being accelerated in fibroblasts overexpressing Rac. Conversely, the activation of the extracellular signal-regulated kinase (Erk1/2) was initially significantly decreased. Subsequent to activation of p38, p53 was activated through serine-15 phosphorylation, and active p53 was translocated from the cytosol to the nucleus. Inhibition of p38 in Rac cells reduced the activation of both p53 and caspase-3. After 60 min in hypertonic medium the rate constants for K+ and taurine efflux were increased, particular in Rac cells. We suggest the following sequence of events in the cell shrinkage-induced apoptotic response: cellular shrinkage activates Rac, with activation of p38, followed by phosphorylation and nuclear translocation of p53, resulting in permeability increases and caspase-3 activation.

Original language	English
Journal	Journal of Physiology
Volume	567
Issue number	2
Pages (from-to)	427-43
Number of pages	16
ISSN	0022-3751
DOIs	https://doi.org/10.1113/jphysiol.2005.087130
Publication status	Published - 2005

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10.1113/jphysiol.2005.087130

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@article{bb115cf0a0f211dd86a6000ea68e967b,

title = "Cell shrinkage as a signal to apoptosis in NIH 3T3 fibroblasts.",

abstract = "Cell shrinkage is a hallmark of the apoptotic mode of programmed cell death, but it is as yet unclear whether a reduction in cell volume is a primary activation signal of apoptosis. Here we studied the effect of an acute elevation of osmolarity (NaCl or sucrose additions, final osmolarity 687 mosmol l(-1)) on NIH 3T3 fibroblasts to identify components involved in the signal transduction from shrinkage to apoptosis. After 1.5 h the activity of caspase-3 started to increase followed after 3 h by the appearance of many apoptotic-like bodies. The caspase-3 activity increase was greatly enhanced in cells expressing a constitutively active G protein, Rac (RacV12A3 cell), indicating that Rac acts upstream to caspase-3 activation. The stress-activated protein kinase, p38, was significantly activated by phosphorylation within 30 min after induction of osmotic shrinkage, the phosphorylation being accelerated in fibroblasts overexpressing Rac. Conversely, the activation of the extracellular signal-regulated kinase (Erk1/2) was initially significantly decreased. Subsequent to activation of p38, p53 was activated through serine-15 phosphorylation, and active p53 was translocated from the cytosol to the nucleus. Inhibition of p38 in Rac cells reduced the activation of both p53 and caspase-3. After 60 min in hypertonic medium the rate constants for K+ and taurine efflux were increased, particular in Rac cells. We suggest the following sequence of events in the cell shrinkage-induced apoptotic response: cellular shrinkage activates Rac, with activation of p38, followed by phosphorylation and nuclear translocation of p53, resulting in permeability increases and caspase-3 activation.",

author = "Friis, {Martin B} and Friborg, {Christel R} and Linda Schneider and Maj-Britt Nielsen and Lambert, {Ian H} and Christensen, {S{\o}ren T} and Hoffmann, {Else K}",

note = "Keywords: Animals; Apoptosis; Caspase 3; Caspases; Cell Membrane Permeability; Cell Size; GTP-Binding Proteins; Mechanotransduction, Cellular; Mice; Mitogen-Activated Protein Kinases; NIH 3T3 Cells; Osmotic Pressure; Tumor Suppressor Protein p53; Water-Electrolyte Balance",

year = "2005",

doi = "10.1113/jphysiol.2005.087130",

language = "English",

volume = "567",

pages = "427--43",

journal = "The Journal of Physiology",

issn = "0022-3751",

publisher = "Wiley-Blackwell",

number = "2",

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TY - JOUR

T1 - Cell shrinkage as a signal to apoptosis in NIH 3T3 fibroblasts.

AU - Friis, Martin B

AU - Friborg, Christel R

AU - Schneider, Linda

AU - Nielsen, Maj-Britt

AU - Lambert, Ian H

AU - Christensen, Søren T

AU - Hoffmann, Else K

N1 - Keywords: Animals; Apoptosis; Caspase 3; Caspases; Cell Membrane Permeability; Cell Size; GTP-Binding Proteins; Mechanotransduction, Cellular; Mice; Mitogen-Activated Protein Kinases; NIH 3T3 Cells; Osmotic Pressure; Tumor Suppressor Protein p53; Water-Electrolyte Balance

PY - 2005

Y1 - 2005

N2 - Cell shrinkage is a hallmark of the apoptotic mode of programmed cell death, but it is as yet unclear whether a reduction in cell volume is a primary activation signal of apoptosis. Here we studied the effect of an acute elevation of osmolarity (NaCl or sucrose additions, final osmolarity 687 mosmol l(-1)) on NIH 3T3 fibroblasts to identify components involved in the signal transduction from shrinkage to apoptosis. After 1.5 h the activity of caspase-3 started to increase followed after 3 h by the appearance of many apoptotic-like bodies. The caspase-3 activity increase was greatly enhanced in cells expressing a constitutively active G protein, Rac (RacV12A3 cell), indicating that Rac acts upstream to caspase-3 activation. The stress-activated protein kinase, p38, was significantly activated by phosphorylation within 30 min after induction of osmotic shrinkage, the phosphorylation being accelerated in fibroblasts overexpressing Rac. Conversely, the activation of the extracellular signal-regulated kinase (Erk1/2) was initially significantly decreased. Subsequent to activation of p38, p53 was activated through serine-15 phosphorylation, and active p53 was translocated from the cytosol to the nucleus. Inhibition of p38 in Rac cells reduced the activation of both p53 and caspase-3. After 60 min in hypertonic medium the rate constants for K+ and taurine efflux were increased, particular in Rac cells. We suggest the following sequence of events in the cell shrinkage-induced apoptotic response: cellular shrinkage activates Rac, with activation of p38, followed by phosphorylation and nuclear translocation of p53, resulting in permeability increases and caspase-3 activation.

AB - Cell shrinkage is a hallmark of the apoptotic mode of programmed cell death, but it is as yet unclear whether a reduction in cell volume is a primary activation signal of apoptosis. Here we studied the effect of an acute elevation of osmolarity (NaCl or sucrose additions, final osmolarity 687 mosmol l(-1)) on NIH 3T3 fibroblasts to identify components involved in the signal transduction from shrinkage to apoptosis. After 1.5 h the activity of caspase-3 started to increase followed after 3 h by the appearance of many apoptotic-like bodies. The caspase-3 activity increase was greatly enhanced in cells expressing a constitutively active G protein, Rac (RacV12A3 cell), indicating that Rac acts upstream to caspase-3 activation. The stress-activated protein kinase, p38, was significantly activated by phosphorylation within 30 min after induction of osmotic shrinkage, the phosphorylation being accelerated in fibroblasts overexpressing Rac. Conversely, the activation of the extracellular signal-regulated kinase (Erk1/2) was initially significantly decreased. Subsequent to activation of p38, p53 was activated through serine-15 phosphorylation, and active p53 was translocated from the cytosol to the nucleus. Inhibition of p38 in Rac cells reduced the activation of both p53 and caspase-3. After 60 min in hypertonic medium the rate constants for K+ and taurine efflux were increased, particular in Rac cells. We suggest the following sequence of events in the cell shrinkage-induced apoptotic response: cellular shrinkage activates Rac, with activation of p38, followed by phosphorylation and nuclear translocation of p53, resulting in permeability increases and caspase-3 activation.

U2 - 10.1113/jphysiol.2005.087130

DO - 10.1113/jphysiol.2005.087130

M3 - Journal article

C2 - 15975986

SN - 0022-3751

VL - 567

SP - 427

EP - 443

JO - The Journal of Physiology

JF - The Journal of Physiology

IS - 2

ER -

Cell shrinkage as a signal to apoptosis in NIH 3T3 fibroblasts.

Abstract

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