Document Detail


Molecular and functional evidence of HCN4 and caveolin-3 interaction during cardiomyocyte differentiation from human embryonic stem cells.
MedLine Citation:
PMID:  23311301     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Maturation of human embryonic stem cell-derived cardiomyocytes (hESC-CM) is accompanied by changes in ion channel expression, with relevant electrophysiological consequences. In rodent CM, the properties of hyperpolarization-activated cyclic nucleotide-gated channel (HCN)4, a major f-channel isoform, depends on the association with caveolin-3 (Cav3). To date, no information exists on changes in Cav3 expression and its associative relationship with HCN4 upon hESC-CM maturation. We hypothesize that Cav3 expression and its compartmentalization with HCN4 channels during hESC-CM maturation accounts for the progression of f-current properties toward adult phenotypes. To address this, hESC were differentiated into spontaneously beating CM and examined at ∼30, ∼60, and ∼110 days of differentiation. Human adult and fetal CM served as references. HCN4 and Cav3 expression and localization were analyzed by real time PCR and immunocyto/histochemistry. F-current was measured in patch-clamped single cells. HCN4 and Cav3 colocalize in adult human atrial and ventricular CM, but not in fetal CM. Proteins and mRNA for Cav3 were not detected in undifferentiated hESC, but expression increased during hESC-CM maturation. At 110 days, HCN4 appeared to be colocalized with Cav3. Voltage-dependent activation of the f-current was significantly more positive in fetal CM and 60-day hESC-CM (midpoint activation, V1/2, ∼ -82 mV) than in 110-day hESC-CM or adult CM (V1/2∼-100 mV). In the latter cells, caveolae disruption reversed voltage dependence toward a more positive or an immature phenotype, with V1/2 at -75 mV, while in fetal CM voltage dependence was not affected. Our data show, for the first time, a developmental change in HCN4-Cav3 association in hESC-CM. Cav3 expression and its association with ionic channels likely represent a crucial step of cardiac maturation.
Authors:
Alexis Bosman; Laura Sartiani; Valentina Spinelli; Martina Del Lungo; Francesca Stillitano; Daniele Nosi; Alessandro Mugelli; Elisabetta Cerbai; Marisa Jaconi
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2013-02-27
Journal Detail:
Title:  Stem cells and development     Volume:  22     ISSN:  1557-8534     ISO Abbreviation:  Stem Cells Dev.     Publication Date:  2013 Jun 
Date Detail:
Created Date:  2013-05-20     Completed Date:  2013-12-11     Revised Date:  2014-06-03    
Medline Journal Info:
Nlm Unique ID:  101197107     Medline TA:  Stem Cells Dev     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1717-27     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Caveolae / metabolism
Caveolin 3 / metabolism*
Cell Differentiation
Cells, Cultured
Cyclic Nucleotide-Gated Cation Channels / metabolism
Electrophysiological Phenomena
Embryonic Stem Cells / physiology*
Humans
Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels / metabolism*
Muscle Proteins / metabolism*
Myocytes, Cardiac / cytology*,  metabolism*
Patch-Clamp Techniques
Potassium Channels / metabolism*
Sarcolemma
Chemical
Reg. No./Substance:
0/CAV3 protein, human; 0/Caveolin 3; 0/Cyclic Nucleotide-Gated Cation Channels; 0/HCN4 protein, human; 0/Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; 0/Muscle Proteins; 0/Potassium Channels
Comments/Corrections

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