Caffeine-induced Ca(2+) signaling as an index of cardiac progenitor cells differentiation

Autor: Gaspare Mostacciuolo, M. Alemanni, Elisa Messina, Lucio Barile, S. Marangoni, Claudia Altomare, Antonio Zaza, Marcella Rocchetti, Alessandro Giacomello
Přispěvatelé: Altomare, C, Barile, L, Marangoni, S, Rocchetti, M, Alemanni, M, Mostacciuolo, G, Giacomello, A, Messina, E, Zaza, A
Rok vydání: 2010
Předmět:
Time Factors
Physiology
cardiac differentiation
Functional response
Mice
Transgenic
Biology
Muscle Development
Ryanodine receptor 2
functional markers
chemistry.chemical_compound
Mice
Adenosine Triphosphate
ca2+ transients
progenitor cells
BIO/09 - FISIOLOGIA
Cell Movement
Physiology (medical)
Caffeine
Animals
Inositol 1
4
5-Trisphosphate Receptors
Myocytes
Cardiac
Calcium Signaling
Progenitor cell
Promoter Regions
Genetic
Cells
Cultured
Ryanodine receptor
Lineage markers
Stem Cells
Troponin I
Promoter
Cell Differentiation
Ryanodine Receptor Calcium Release Channel
Ca transients
Cardiac differentiation
Progenitor cells
Functional markers
Myocardial Contraction
Electric Stimulation
Cell biology
Mice
Inbred C57BL
chemistry
Gene Expression Regulation
Immunology
cardiovascular system
Female
Cardiology and Cardiovascular Medicine
Biomarkers
Explant culture
Zdroj: Basic research in cardiology. 105(6)
ISSN: 1435-1803
Popis: Cardiac progenitor cells (CPCs), migrating from heart tissue, in culture aggregate to form cardiospheres (CSs) in which replication and cardiogenic differentiation occur. However, the frequency of functional differentiation in CSs and the role of cell clustering in supporting it remain to be established. The aim of our study is to quantify differentiation of a muscle-type Ca(2+) release mechanism in CS-derived cells, correlate it with cardiac differentiation markers and test its dependency on CS formation. CPCs migrating from murine cardiac explants were studied prior and after CSs formation (Pre-CS and Post-CS). Inducibility of RyR- and IP3-R-mediated Ca(2+) transients in individual cells was tested by exposure to caffeine and ATP, respectively; expression of cardiac and non-cardiac lineage markers was assessed. Caffeine responsiveness was negligible in Pre-CS cells and increased by 7.5 fold in Post-CS cells (3.6 vs. 26.9%; p < 0.05), and was closely correlated with activation of the cardiac TnI gene promoter. ATP-induced responses, frequent in Pre-CS (86%), were slightly increased in Post-CS cells (94%; p < 0.05). Expression of cardiac-specific Ca(2+)-handling proteins (Cav1.2, NCX1, RyR2, SERCA2a) was either limited to the Post-CS stage, or markedly enhanced. CS beating was infrequent, but its pharmacology was compatible with cardiac excitation-contraction coupling. Expression of non-cardiac lineage was low in general, and similar between Pre- and Post-CS cells. Culture conditions inhibiting CSs formation prevented the increase in caffeine responders. In conclusion, clustering in CSs leads to the induction of a muscle-specific functional response in about 30% of CPCs; this is accompanied by development of a cardiac-specific expression pattern.
Databáze: OpenAIRE
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