Role of estrogen receptors, PKC and Src in ERK2 and p38 MAPK signaling triggered by 17β-estradiol in skeletal muscle cells

Ronda, Ana Carolina; Buitrago, Claudia; Boland, Ricardo

doi:10.1016/j.jsbmb.2010.05.002

Cited by 29 publications

(28 citation statements)

References 60 publications

(88 reference statements)

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“…We have previously observed that E2 through ERα stimulates ERK phosphorylation in C2C12 cells [24,25]. We have now investigated where this intracellular phosphorylation takes place and whether the hormone induces mobilization of the MAPK.…”

Section: Resultsmentioning

confidence: 99%

“…To this end, C2C12 cells were incubated with the specific mitochondrial probe Mitotracker (red fluorescence). Then myoblasts were treated with E2 or its vehicle for 15 min when maximal phosphorylation of ERK by the estrogen has been shown to occur [24,25]. As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…This event prevents both the decrease in COXIV activity and the loss of mitochondrial membrane potential induced by hydrogen peroxide in C2C12 myoblasts. We have previously demonstrated that E2 induces a fast stimulation of ERK through ERα [24,25]. We also showed that the hormone protects the cells against apoptosis induced by H 2 O 2 through the mitochondrial pathway and involving ERK activation [9,10,11].…”

Section: Discussionmentioning

confidence: 99%

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17�-Estradiol Protects Mitochondrial Functions through Extracellular-Signal-Regulated Kinase in C2C12 Muscle Cells

Ronda

Vasconsuelo²,

Boland³

2013

Cell Physiol Biochem

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Background/Aims: We have previously shown that exposure to 17β-estradiol (E2) prior to induction of apoptosis with H₂O₂ protects skeletal muscle cells against oxidative damage. However, the mechanism involved in the protective action of the hormone is poorly understood. In the present study, we focused on the mechanism by which ERK mediates this survival effect in connection with COXIV activity and mitochondrial membrane potential. Methods: Immunocytochemistry, Western blot, cytochrome c oxidase complex IV (COXIV) activity, coimmunoprecipitation and JC-1 dye by flow cytometry were carried out using C2C12 myoblasts as experimental model. Results: E2 is able to activate ERK and then induces its translocation to mitochondria. Using the pharmacological inhibitor of ERK activation U0126 we show that E2, through ERK activation, is able to enhance COXIV activity. Moreover, the hormone increases the interaction between COXIV and ERK. Also, we found that hydrogen peroxide decreases COXIV activity and that preincubation of the cells with E2 prior to induction of apoptosis prevents this effect. In addition, we observe that the estrogen inhibits the collapse of mitochondrial membrane potential induced by H₂O₂, involving ERK and COXIV. Conclusion: Our data demonstrate that E2 promotes ERK activation and translocation to mitochondria preventing the decline in COXIV activity and in turn, alteration of mitochondrial membrane potential by oxidative stress, in C2C12 myoblasts.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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17�-Estradiol Protects Mitochondrial Functions through Extracellular-Signal-Regulated Kinase in C2C12 Muscle Cells

Ronda

Vasconsuelo²,

Boland³

2013

Cell Physiol Biochem

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“…Importantly, when such shifts are detected they are modest, in the magnitude of 10% or less, and thus likely have little impact on the extent of force potentiation reported here. E 2 activates several kinases and signaling pathways in various skeletal muscle cell lines (4,8,48) and skeletal muscle of rodents (32,56,68). In our screen of E 2 -responsive kinases using a kinase inhibitor library and C 2 C 12 cells, we identified 11 inhibitors that decreased pRLC triggered by E 2 ( Table 1).…”

Section: Discussionmentioning

confidence: 99%

“…For example, ERs have been implicated in the regulation of gene transcription in myoblasts, including kinases that regulate GLUT4, myogenin, and myosin heavy chain expression (12). E 2 is also an important molecule activating several kinases and kinase cascades, such as phosphatidylinositide 3-kinase/Akt (PI3K/Akt), MAPK, and AMP-activated protein kinase (AMPK), in a multitude of cells and tissues including skeletal muscle (46,48,68). These cellular processes result in altered kinase activity and thus protein phosphorylation.…”

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confidence: 99%

Estradiol modulates myosin regulatory light chain phosphorylation and contractility in skeletal muscle of female mice

Lai

Collins

Colson

et al. 2016

American Journal of Physiology-Endocrinology and Metabolism

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-Impairment of skeletal muscle function has been associated with changes in ovarian hormones, especially estradiol. To elucidate mechanisms of estradiol on skeletal muscle strength, the hormone's effects on phosphorylation of the myosin regulatory light chain (pRLC) and muscle contractility were investigated, hypothesizing an estradiol-specific beneficial impact. In a skeletal muscle cell line, C 2C12, pRLC was increased by 17␤-estradiol (E 2) in a concentration-dependent manner. In skeletal muscles of C57BL/6 mice that were E 2 deficient via ovariectomy (OVX), pRLC was lower than that from ovary-intact, sham-operated mice (Sham). The reduced pRLC in OVX muscle was reversed by in vivo E 2 treatment. Posttetanic potentiation (PTP) of muscle from OVX mice was low compared with that from Sham mice, and this decrement was reversed by acute E 2 treatment, demonstrating physiological consequence. Western blot of those muscles revealed that low PTP corresponded with low pRLC and higher PTP with greater pRLC. We aimed to elucidate signaling pathways affecting E 2-mediated pRLC using a kinase inhibitor library and C 2C12 cells as well as a specific myosin light chain kinase inhibitor in muscles. PI3K/Akt, MAPK, and CamKII were identified as candidate kinases sensitive to E 2 in terms of phosphorylating RLC. Applying siRNA strategy in C 2C12 cells, pRLC triggered by E 2 was found to be mediated by estrogen receptor-␤ and the G protein-coupled estrogen receptor. Together, these results provide evidence that E 2 modulates myosin pRLC in skeletal muscle and is one mechanism by which this hormone can affect muscle contractility in females. estrogen; estrogen receptor; kinase; post tetanic potentiation; RLC OVARIAN HORMONE DEFICIENCY is related to muscle dysfunction and loss of strength. For example, declines in strength are accelerated around the time of menopause (35,41,51), and skeletal muscles of postmenopausal women on estrogen-based hormone therapy are stronger than those not on hormone therapy (10,17,49). In mouse models, contractile characteristics are impaired in muscles of ovariectomized compared with sham-operated mice (16,38,39,53,65). It was suggested (41) and subsequently demonstrated that ovarian hormones influence muscle strength by directly affecting the function of contractile proteins (43,66), particularly the structure and function of myosin (38,39). Among the ovarian hormones, 17␤-estradiol (E 2 ) appears to be the key hormone, because it reverses muscle weakness and contractile protein dysfunction caused by ovariectomy. Specifically, reduced muscle and contractile protein functions were restored when E 2 was administered to ovariectomized mice (16,38,53). In a chemical model of premature aging, of which the ovaries of young adult mice were locally and specifically induced to be senescent, E 2 treatment improved some muscle contractile functions (18). Although these studies have demonstrated that E 2 impacts muscle and myosin functions, the molecular mechanisms by which this hormone affects the structure a...

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Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells

Ronda

Boland

2015

J. Cell. Biochem.

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G-protein-coupled receptor 30 (GPR30) is an estrogen receptor that initiates several rapid, non-genomic signaling events triggered by E2. GPR30 has recently been identified in C2C12 cells; however, little is known about the intracelular distribution and its role in C2C12 myoblasts and myotubes. By western blotting and immunohistochemistry, we evidenced expression of GPR30. While in C2C12 myoblasts, the receptor was present in nucleus, mitochondria, and endoplasmic reticulum, in C2C12 myotubes, it was additionally found in cytoplasm. Using trypan blue uptake assay to determine cellular death and fluorescent microscopy to evaluate picnotic nuclei and mitochondrial distribution, we demonstated that treatment of C2C12 myoblasts with G1 (GPR30 agonist) did not protect the cells against apoptosis induced by H2O2 as E2. However, when G15 (GPR30 antagonist) was used, E2 could not prevent the damage caused by the oxidative stress. Further, some of the molecular mechanisms involved were investigated by wertern blot assays. Thus, E2 was able to induce AKT phosphorylation in apoptotic conditions and ERK phosphorylation in proliferating C2C12 cells but not when the cultures were incubated with G15. Additionally, using G15 antagonist we have found that GPR30 participates in the myogenin expression and creatine kinase activity stimulated by E2 in the first steps of C2C12 differentiation. Althogether these findings provide evidences showing that GPR30 is expressed in diverse intracellular compartments in undifferentiated and differentiated C2C12 cells and mediates E2 actions.

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Role of estrogen receptors, PKC and Src in ERK2 and p38 MAPK signaling triggered by 17β-estradiol in skeletal muscle cells

Cited by 29 publications

References 60 publications

17�-Estradiol Protects Mitochondrial Functions through Extracellular-Signal-Regulated Kinase in C2C12 Muscle Cells

17�-Estradiol Protects Mitochondrial Functions through Extracellular-Signal-Regulated Kinase in C2C12 Muscle Cells

Estradiol modulates myosin regulatory light chain phosphorylation and contractility in skeletal muscle of female mice

Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells

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