Measurement of oxidant-induced signal transduction proteins using cell imaging

Poynter, Matthew E.; Janssen–Heininger, Yvonne M. W.; Buder-Hoffmann, Sylke A.; Taatjes, Douglas J.; Mossman, Brooke T.

doi:10.1016/s0891-5849(99)00202-6

Cited by 15 publications

(4 citation statements)

References 7 publications

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“…Further analysis of the requirement for ROS in anti-CD3–stimulated signal transduction focused upon the MAP kinase family, which have been shown to be sensitive to both exogenous and endogenously generated ROS ( 31 , 32 ). To measure MAP kinase activation in the transiently transfected cell systems, a protocol for immunohistochemical staining of cells with an antibody specific for phosphorylated ERK1/2 ( 22 ) was adapted to lymphocytes transfected with GFP. Antibody specificity was demonstrated by competition of binding with a phosphorylated ERK protein supplied by the manufacturers, and by incubation with the MEK1/2 inhibitor PD98059 ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Cryopreservation did not appear to affect cells as measured by GFP staining of cells. The cryopreserved cells were thawed and immunochemically stained for MAP kinase activation essentially as described previously ( 22 ). Cells were washed with wash buffer (PBS containing 1% BSA and 0.1% Triton X-100) and blocked with 2% nonfat dry milk at room temperature for 30 min.…”

Section: Methodsmentioning

confidence: 99%

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Discrete Generation of Superoxide and Hydrogen Peroxide by T Cell Receptor Stimulation

Devadas

Zaritskaya

Rhee

et al. 2002

The Journal of Experimental Medicine

417

153

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Receptor-stimulated generation of reactive oxygen species (ROS) has been shown to regulate signal transduction, and previous studies have suggested that T cell receptor (TCR) signals may involve or be sensitive to ROS. In this study, we have shown for the first time that TCR cross-linking induced rapid (within 15 min) generation of both hydrogen peroxide and superoxide anion, as defined with oxidation-sensitive dyes, selective pharmacologic antioxidants, and overexpression of specific antioxidant enzymes. Furthermore, the data suggest the novel observation that superoxide anion and hydrogen peroxide are produced separately by distinct TCR-stimulated pathways. Unexpectedly, TCR-stimulated activation of the Fas ligand (FasL) promoter and subsequent cell death was dependent upon superoxide anion, but independent of hydrogen peroxide, while nuclear factor of activated T cells (NFAT) activation or interleukin 2 transcription was independent of all ROS. Anti-CD3 induced phosphorylation of extracellular signal–regulated kinase (ERK)1/2 required hydrogen peroxide generation but was unaffected by superoxide anion. Thus, antigen receptor signaling induces generation of discrete species of oxidants that selectively regulate two distinct redox sensitive pathways, a proapoptotic (FasL) and a proliferative pathway (ERK).

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Discrete Generation of Superoxide and Hydrogen Peroxide by T Cell Receptor Stimulation

Devadas

Zaritskaya

Rhee

et al. 2002

The Journal of Experimental Medicine

417

153

View full text Add to dashboard Cite

show abstract

“…Most of the molecules involved in the mechanisms of signal transduction can be labelled using immunocytochemical methods. However, none of these methods allows specific identification of the biochemical event resulting from the stressful stimulus and none has great clinical importance 33 …”

Section: Evaluation Of Oxidative Stressmentioning

confidence: 99%

Antioxidants in dermocosmetology: from the laboratory to clinical application

Andreassi¹,

Andreassi

2003

J of Cosmetic Dermatology

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Oxygen situated in cutaneous cells can be activated by light. This makes the integumentary apparatus particularly vulnerable to oxidative damage and is responsible for the immediate cutaneous damage that is the basis of late phenomena, such as photo-induced ageing and tumours. Thus, the cosmetic industry has undertaken research and development into antioxidant-based products able to protect the skin from the effect of pro-oxidizing noxae. This review re-examines both antioxidants suitable for dermatological application and skin care products with antioxidant capacity, as well as the laboratory methods used to evaluate the effects and in vivo efficacy of antioxidants.

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“…Mitogen‐activated protein kinases (MAPKs) consist of three family members: the extracellular signal‐regulated kinases (ERK), JNK, and p38 MAPK [Raman et al, 2007]. ERK is generally considered to be a survival factor involved in the protective actions of growth factors in apoptosis [Walker et al, 1998; Poynter et al, 1999], whereas p38 and JNK are usually required for induction of apoptosis by diverse stimuli [Nagai et al, 2007]. However, the individual roles of JNK and p38 remain to be clarified in different cell types and conditions.…”

mentioning

confidence: 99%

LPA rescues ER stress‐associated apoptosis in hypoxia and serum deprivation‐stimulated mesenchymal stem cells

Hua

Liu

et al. 2010

J of Cellular Biochemistry

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Poor viability of transplanted mesenchymal stem cells (MSCs) in the infracted heart has limited their therapeutic efficacy in cardiac repair after myocardial infarction. We previously demonstrated that hypoxia and serum deprivation (hypoxia/SD) induced mitochondria-dependent apoptosis in MSCs, while lysophosphatidic acid (LPA) could almost completely block this apoptotic process. However, the role of endoplasmic reticulum (ER) stress and its upstream signaling events in hypoxia/SD-induced MSC apoptosis remain largely unknown. Here we found that hypoxia/SD-induced MSC apoptosis was associated with ER stress, as shown by the induction of CHOP expression and procaspase-12 cleavage, while the effects were abrogated by LPA treatment, suggesting ER stress is also a target of LPA. Furthermore, hypoxia/SD induced p38 activation, inhibition of which resulted in decreases of apoptotic cells, procaspase-12 cleavage and mitochondrial cytochrome c release that function in parallel in MSC apoptosis. Unexpectedly, p38 inhibition enhanced hypoxia/SD-induced CHOP expression. Interestingly, p38 activation, a common process mediating various biological effects of LPA, was inhibited by LPA in this study, and the regulation of p38 pathway by LPA was dependent on LPA(1/3)/Gi/ERK1/2 pathway-mediated MKP-1 induction but independent of PI3K/Akt pathway. Collectively, our findings indicate that ER stress is a target of LPA to antagonize hypoxia/SD-induced MSC apoptosis, and the modulation of mitochondrial and ER stress-associated apoptotic pathways by LPA is at least partly dependent on LPA(1/3)/Gi/ERK/MKP-1 pathway-mediated p38 inhibition. This study may provide new anti-apoptotic targets for elevating the viability of MSCs for therapeutic potential of cardiac repair.

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Measurement of oxidant-induced signal transduction proteins using cell imaging

Cited by 15 publications

References 7 publications

Discrete Generation of Superoxide and Hydrogen Peroxide by T Cell Receptor Stimulation

Discrete Generation of Superoxide and Hydrogen Peroxide by T Cell Receptor Stimulation

Antioxidants in dermocosmetology: from the laboratory to clinical application

LPA rescues ER stress‐associated apoptosis in hypoxia and serum deprivation‐stimulated mesenchymal stem cells

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