scholarly journals Apocynin and Diphenyleneiodonium Induce Oxidative Stress and Modulate PI3K/Akt and MAPK/Erk Activity in Mouse Embryonic Stem Cells

2016 ◽  
Vol 2016 ◽  
pp. 1-14 ◽  
Author(s):  
Jan Kučera ◽  
Lucia Binó ◽  
Kateřina Štefková ◽  
Josef Jaroš ◽  
Ondřej Vašíček ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Signaling Pathways ◽  
Intracellular Signaling ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells ◽  
Cellular Functions ◽  
Differentiation Status ◽  
Canonical Wnt ◽  
Erk Activity

Reactive oxygen species (ROS) are important regulators of cellular functions. In embryonic stem cells, ROS are suggested to influence differentiation status. Regulated ROS formation is catalyzed primarily by NADPH-dependent oxidases (NOXs). Apocynin and diphenyleneiodonium are frequently used inhibitors of NOXs; however, both exhibit uncharacterized effects not related to NOXs inhibition. Interestingly, in our model of mouse embryonic stem cells we demonstrate low expression of NOXs. Therefore we aimed to clarify potential side effects of these drugs. Both apocynin and diphenyleneiodonium impaired proliferation of cells. Surprisingly, we observed prooxidant activity of these drugs determined by hydroethidine. Further, we revealed that apocynin inhibits PI3K/Akt pathway with its downstream transcriptional factor Nanog. Opposite to this, apocynin augmented activity of canonical Wnt signaling. On the contrary, diphenyleneiodonium activated both PI3K/Akt and Erk signaling pathways without affecting Wnt. Our data indicates limits and possible unexpected interactions of NOXs inhibitors with intracellular signaling pathways.

scholarly journals Canonical Wnt Signaling Induces a Primitive Endoderm Metastable State in Mouse Embryonic Stem Cells

Stem Cells ◽  
2013 ◽  
Vol 31 (4) ◽  
pp. 752-764 ◽  
Author(s):  
Feodor D. Price ◽  
Hang Yin ◽  
Andrew Jones ◽  
Wilfred van Ijcken ◽  
Frank Grosveld ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Wnt Signaling ◽  
Metastable State ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells ◽  
Primitive Endoderm ◽  
Canonical Wnt Signaling ◽  
Canonical Wnt

62 EFFECT OF PROGESTERONE ON CALCIUM REGULATION DURING DIFFERENTIATION OF MOUSE EMBRYONIC STEM CELLS INTO CARDIOMYOCYTES

2016 ◽  
Vol 28 (2) ◽  
pp. 161
Author(s):  
Y.-K. Choi ◽  
H. Y. Kang ◽  
J.-U. Hwang ◽  
T. D. Nam ◽  
E.-B. Jeung
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Calcium Regulation ◽  
Mouse Embryonic Stem Cells ◽  
Cardiac Differentiation ◽  
Mrna Levels ◽  
Cellular Functions ◽  
Skeletal Muscle Contraction ◽  
Cardiac Lineage

During spontaneous cardiac differentiation of mouse embryonic stem cells (mESCs, cell line E14), the effect of progesterone on calcium regulation was investigated. Calcium (Ca2+) release from sarcoplasmic reticulum (SR) regulates various cellular functions including the smooth or skeletal muscle contraction. The cardiac L-type Ca2+ channel plays a key role in excitation-contraction coupling of cardiomyocytes and contraction-related gene expression. The mESCs formed mouse embryonic bodies (mEBs) by hanging-drop for 4 days, and mEBs were suspended for 2 days in differentiation medium; DMEM/F:12, 15% charcoal-dextran-treated FBS, β-mercaptoethanol, minimal essential medium NEAA, and penicillin/streptomycin. Then, mEBs were attached onto 6-well culture plates and differentiated into cardiomyocytes. We analysed mRNA expressions for the cardiac lineage markers and calcium-regulating genes. Percentage of beating mEBs was time-dependently increased during differentiation. Differentiated mEBs showed the highest beating ratio (92.11 ± 2.98%) after attachment for 12 days. Beating ratio was decreased to 64.86 ± 4.25% in progesterone-treated mEBs. The mRNA levels of cardiac markers such as Tbx20, Isl1, Foxh1, cTn1, and RyR2 were increased, and troponin protein was observed in beating mEBs via immunocytochemistry. Expression of calcium/contraction regulating genes including Trpv2, Ryr2, CaM2, and Mlck3 was down-regulated by progesterone treatment. These results indicate that progesterone has influences on cardiac differentiation and contraction of cardiomyocytes through regulating intercellular calcium ion. This research was supported by a grant (15182MFDS460) from the Ministry of Food and Drug Safety in 2015.

Functional expression of Ca2+ signaling pathways in mouse embryonic stem cells

Cell Calcium ◽  
2004 ◽  
Vol 36 (2) ◽  
pp. 135-146 ◽  
Author(s):  
Eri Yanagida ◽  
Satoshi Shoji ◽  
Yoshiyuki Hirayama ◽  
Fumio Yoshikawa ◽  
Keishi Otsu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Signaling Pathways ◽  
Embryonic Stem ◽  
Functional Expression ◽  
Mouse Embryonic Stem Cells
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