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Functional expression of Ca2+ signaling pathways in mouse embryonic stem cells
Eri Yanagida1, Satoshi Shoji, Yoshiyuki Hirayama
1Department of Cardiovascular Diseases, Medical Research Institute, Tokyo Medical and Dental University, Yushima, Bunkyo-ku, Japan.
Cell Calcium
|June 15, 2004
Summary
Undifferentiated mouse embryonic stem cells utilize inositol-1,4,5-triphosphate receptors (InsP3Rs) for calcium release and store-operated calcium channels (SOCs) for calcium entry, crucial for maintaining low intracellular calcium levels.
Area of Science:
- Cell Biology
- Stem Cell Biology
- Calcium Signaling
Background:
- Mouse embryonic stem (mES) cells possess pluripotency but their calcium (Ca2+) signaling physiology remains incompletely understood.
- Investigating Ca2+ signaling is vital for understanding mES cell function and differentiation potential.
Purpose of the Study:
- To elucidate the Ca2+ signaling pathways, including release, entry, and extrusion mechanisms, in undifferentiated mES cells.
- To identify the specific channels and transporters involved in regulating intracellular Ca2+ concentration ([Ca2+]i) in mES cells.
Main Methods:
- Confocal Ca2+ imaging to monitor intracellular Ca2+ dynamics.
- Patch clamp techniques to record ion channel activity.
- RT-PCR to detect gene expression of Ca2+ handling proteins.
Main Results:
- ATP and histamine triggered Ca2+ release via inositol-1,4,5-triphosphate receptors (InsP3Rs).
- Capacitative Ca2+ entry (CCE) and store-operated Ca2+ currents were observed upon store depletion.
- Both Na+/Ca2+ exchangers (NCXs) and plasma membrane Ca2+ pumps (PMCAs) were identified as key Ca2+ extrusion systems.
Conclusions:
- Ca2+ release in mES cells is primarily mediated by InsP3Rs.
- Store-operated channels (SOCs) are the main pathway for Ca2+ entry.
- NCXs and PMCAs are crucial for maintaining low basal [Ca2+]i in mES cells.