Dataset Information

Ryanodine receptor RyR1-mediated elevation of Ca²⁺ concentration is required for the late stage of myogenic differentiation and fusion.

ABSTRACT:

Background

Cytosolic Ca²⁺ plays vital roles in myogenesis and muscle development. As a major Ca²⁺ release channel of endoplasmic reticulum (ER), ryanodine receptor 1 (RyR1) key mutations are main causes of severe congenital myopathies. The role of RyR1 in myogenic differentiation has attracted intense research interest but remains unclear.

Results

In the present study, both RyR1-knockdown myoblasts and CRISPR/Cas9-based RyR1-knockout myoblasts were employed to explore the role of RyR1 in myogenic differentiation, myotube formation as well as the potential mechanism of RyR1-related myopathies. We observed that RyR1 expression was dramatically increased during the late stage of myogenic differentiation, accompanied by significantly elevated cytoplasmic Ca²⁺ concentration. Inhibition of RyR1 by siRNA-mediated knockdown or chemical inhibitor, dantrolene, significantly reduced cytosolic Ca²⁺ and blocked multinucleated myotube formation. The elevation of cytoplasmic Ca²⁺ concentration can effectively relieve myogenic differentiation stagnation by RyR1 inhibition, demonstrating that RyR1 modulates myogenic differentiation via regulation of Ca²⁺ release channel. However, RyR1-knockout-induced Ca²⁺ leakage led to the severe ER stress and excessive unfolded protein response, and drove myoblasts into apoptosis.

Conclusions

Therefore, we concluded that Ca²⁺ release mediated by dramatic increase in RyR1 expression is required for the late stage of myogenic differentiation and fusion. This study contributes to a novel understanding of the role of RyR1 in myogenic differentiation and related congenital myopathies, and provides a potential target for regulation of muscle characteristics and meat quality.

SUBMITTER: Qiu K

PROVIDER: S-EPMC8832842 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature

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Publications

Ryanodine receptor RyR1-mediated elevation of Ca<sup>2+</sup> concentration is required for the late stage of myogenic differentiation and fusion.

Qiu Kai K Wang Yubo Y Xu Doudou D He Linjuan L Zhang Xin X Yan Enfa E Wang Lu L Yin Jingdong J

Journal of animal science and biotechnology 20220211 1

<h4>Background</h4>Cytosolic Ca<sup>2+</sup> plays vital roles in myogenesis and muscle development. As a major Ca<sup>2+</sup> release channel of endoplasmic reticulum (ER), ryanodine receptor 1 (RyR1) key mutations are main causes of severe congenital myopathies. The role of RyR1 in myogenic differentiation has attracted intense research interest but remains unclear.<h4>Results</h4>In the present study, both RyR1-knockdown myoblasts and CRISPR/Cas9-based RyR1-knockout myoblasts were employed t ...[more]

PMID: 35144690

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Project description:Mucociliary clearance is a pivotal physiological mechanism that protects the lung by cleaning the airways from pollution and colonization, thereby preventing infection. Ciliary function is influenced by various signal transduction cascades, and Ca2+ represents a key second messenger. A fixed 20:1 combination of cafedrine and theodrenaline has been widely used to treat perioperative hypotension and emergency hypotensive states since the 1960s; however, its effect on the intracellular Ca2+ concentration ([Ca2+]i) of respiratory epithelium remains unknown. Therefore, human tracheal epithelial cells were exposed to the clinically applied 20:1 mixture of cafedrine/theodrenaline and the individual substances separately. [Ca2+]i was assessed by FURA-2 340/380 fluorescence ratio. Pharmacological inhibitors were applied to elucidate relevant signal transduction cascades, and reverse transcription polymerase chain reaction (RT-PCR) was performed on murine tracheal epithelium to analyze ryanodine receptor (RyR) subtype expression. All three pharmacological preparations instantaneously induced a steep increase in [Ca2+]i that quickly returned to its baseline value despite the persistence of each substance. Peak [Ca2+]i following the administration of 20:1 cafedrine/theodrenaline, cafedrine alone, and theodrenaline alone increased in a dose-dependent manner, with median effective concentrations of 0.35 mM (7.32 mM cafedrine and 0.35 mM theodrenaline), 3.14 mM, and 3.45 mM, respectively. When extracellular Ca2+ influx was inhibited using a Ca2+-free buffer solution, the peak [Ca2+]i following the administration of cafedrine alone and theodrenaline alone were reduced but not abolished. No alteration in [Ca2+]i compared with baseline [Ca2+]i was observed during β-adrenergic receptor inhibition. Depletion of caffeine-sensitive stores and inhibition of RyR, but not IP3 receptors, completely abolished any increase in [Ca2+]i. However, [Ca2+]i still increased following the depletion of mitochondrial Ca2+ stores using 2,4-dinitrophenol. RT-PCR revealed RyR-2 and RyR-3 expression on murine tracheal epithelium. Although our experiments showed that cafedrine/theodrenaline, cafedrine alone, or theodrenaline alone release Ca2+ from intracellular stores through mechanisms that are exclusively triggered by β-adrenergic receptor stimulation, which most probably lead to RyR activation, clinical plasma concentrations are considerably lower than those used in our experiments to elicit an increase in [Ca2+]i; therefore, further studies are needed to evaluate the ability of cafedrine/theodrenaline to alter mucociliary clearance in clinical practice.

Dataset Information

Ryanodine receptor RyR1-mediated elevation of Ca²⁺ concentration is required for the late stage of myogenic differentiation and fusion.

Background

Results

Conclusions

Publications

Ryanodine receptor RyR1-mediated elevation of Ca<sup>2+</sup> concentration is required for the late stage of myogenic differentiation and fusion.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Dataset Information

Ryanodine receptor RyR1-mediated elevation of Ca2+ concentration is required for the late stage of myogenic differentiation and fusion.

Background

Results

Conclusions

Publications

Ryanodine receptor RyR1-mediated elevation of Ca<sup>2+</sup> concentration is required for the late stage of myogenic differentiation and fusion.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Ryanodine receptor RyR1-mediated elevation of Ca²⁺ concentration is required for the late stage of myogenic differentiation and fusion.