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Modified Mouse Embryonic Stem Cell based Assay for Quantifying Cardiogenic Induction Efficiency
Published on: April 22, 2011
Mouse embryonic stem cell-derived cardiomyocytes express functional adrenoceptors
Hongtao Yang1, Yongzhen Zhang, Zhiqiang Liu
1Department of Cardiology, Peking University Third Hospital, Beijing 100083, China.
Abstract:
The cardiogenic capacity of embryonic stem (ES) cells has been well-investigated. However, little is known about the development of adrenoceptor (AR) systems during the process of ES cell differentiation, which are critically important in cardiac physiology and pharmacology. In this present study, we investigated the expression profile of adrenoceptor subtypes, beta-adrenergic modulation of muscarinic receptors and adrenoceptor-related signaling in cardiomyocytes derived from ES cells (ESCMs). Reverse transcription-polymerase chain reaction revealed that undifferentiated mouse ES cells expressed alpha(1A)-, alpha(1B)-, alpha(1D)- and beta(2)-AR mRNA. However, beta(1)-AR was only expressed after vitamin C induction. The expressions of alpha(1A)-, alpha(1D)- and beta(1)-ARs increased significantly while alpha(1B)- and beta(2)-ARs showed no significant change during the differentiation process. Furthermore, we detected the expression of tyrosine hydroxylase. Both alpha(1)-AR and beta-AR could activate extracellular responsive kinase in ESCMs. Isoprenaline could inhibit the expression of M(2) muscarinic receptor protein. CGP20712A, a beta(1)-AR antagonist, up-regulated the expression of M(2) muscarinic receptor while ICI118551, a beta(2)-AR antagonist, showed no effect. These results indicated that functional adrenoceptors and tyrosine hydroxylase, a critical enzyme in catecholamine biosynthesis, were differentially expressed in ESCMs. Adrenoceptor-related signaling pathways and beta-adrenergic modulation of muscarinic receptors were established during differentiation.
Insights
Embryonic stem cell-derived cardiomyocytes (ESCMs) develop functional adrenoceptors (ARs) and related signaling pathways during differentiation. Beta-adrenergic modulation of muscarinic receptors is also established, impacting cardiac physiology.
Area of Science:
- Cardiology
- Stem Cell Biology
- Pharmacology
Background:
- Embryonic stem cell (ES) cardiogenesis is well-studied, but adrenoceptor (AR) system development during differentiation remains unclear.
- AR systems are crucial for cardiac physiology and pharmacology.
- Understanding AR development in ES cell-derived cardiomyocytes (ESCMs) is vital.
Purpose of the Study:
- To investigate adrenoceptor subtype expression during ES cell differentiation into cardiomyocytes.
- To examine beta-adrenergic modulation of muscarinic receptors in ESCMs.
- To analyze adrenoceptor-related signaling pathways in ESCMs.
Main Methods:
- Reverse transcription-polymerase chain reaction (RT-PCR) for mRNA expression analysis.
- Vitamin C induction for differentiation.
- Pharmacological manipulation using AR antagonists (CGP20712A, ICI118551) and agonists (Isoprenaline).
Main Results:
- Undifferentiated ES cells expressed alpha(1A)-, alpha(1B)-, alpha(1D)-, and beta(2)-AR mRNA; beta(1)-AR emerged post-differentiation.
- Alpha(1A)-, alpha(1D)-, and beta(1)-AR expression increased significantly during differentiation.
- Both alpha(1)-AR and beta-AR activated extracellular signal-regulated kinase (ERK); Isoprenaline inhibited M(2) muscarinic receptor protein expression.
Conclusions:
- Functional adrenoceptors and tyrosine hydroxylase are differentially expressed during ESCM differentiation.
- Adrenoceptor signaling pathways and beta-adrenergic modulation of muscarinic receptors are established in ESCMs.
- These findings provide insights into the development of adrenergic signaling in the embryonic heart.
