Changes in intracellular Ca2+ concentration ([Ca2+]i) play a central role in neuronal differentiation. However, Ca2+ signaling in this process remains poorly understood and it is unknown whether embryonic and adult stem cells share the same signaling pathways. To clarify this issue, neuronal differentiation was analyzed in two cell lines: embryonic P19 carcinoma stem cells (CSCs) and adult murine bone-marrow mesenchymal stem cells (MSC). We studied Ca2+ release from the endoplasmic reticulum via intracellular ryanodine-sensitive (RyR) and IP3-sensitive (IP3R) receptors. We observed that caffeine, a RyR agonist, induced a [Ca2+]i response that increased throughout neuronal differentiation. We also demonstrated a functional coupling between RyRs and L- but not with N-, P-, or Q-type Cav1 Ca2+ channels, both in embryonal CSC and adult MSC. We also found that agonists of L-type channels and of RyRs increase neurogenesis and neuronal differentiation, while antagonists of these channels have the opposite effect. Thus, our data demonstrate that in both cell lines RyRs control internal Ca2+ release following voltage-dependent Ca2+ entry via L-type Ca2+ channels. This study shows that both in embryonal CSC and adult MSC [Ca2+]i is controlled by a common pathway, indicating that coupling of L-type Ca2+ channels and RyRs may be a conserved mechanism necessary for neuronal differentiation.
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