Abstract
We have used P19 embryonal carcinoma cells as in vitro model for early neurogenesis to study ionotropic P2X and metabotropic P2Y receptor-induced Ca(2+) transients and their participation in induction of proliferation and differentiation. In embryonic P19 cells, P2Y(1), P2Y(2) and P2X(4) receptors or P2X-heteromultimers with similar P2X(4) pharmacology were responsible for ATP and ATP analogue-induced Ca(2+) transients. In neuronal-differentiated cells, P2Y(2,) P2Y(6), P2X(2) and possibly P2X(2)/P2X(6) heteromeric receptors were the major mediators of the elevations in intracellular free calcium concentration [Ca(2+)](i). We have collected evidence for the involvement of metabotropic purinergic receptors in proliferation induction of undifferentiated and neural progenitor cells by using a BrdU-incorporation assay. ATP-, UTP-, ADP-, 2-MeS-ATP- and ADP-betaS-induced proliferation in P19 cells was mediated by P2Y(1) and P2Y(2) receptors as judged from pharmacological profiles of receptor responses. ATP-provoked acceleration of neuronal differentiation, determined by analysis of nestin and neuron-specific enolase gene and protein expression, also resulted from P2Y(1) and P2Y(2) receptor activation. Proliferation- and differentiation-induction involved the activation of inositol-trisphosphate sensitive intracellular Ca(2+) stores.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenosine Triphosphate / analogs & derivatives
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Adenosine Triphosphate / pharmacology
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Calcium / metabolism
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Calcium Signaling / physiology
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Cell Line, Tumor
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Cell Proliferation / drug effects*
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Embryonal Carcinoma Stem Cells
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Embryonic Induction / drug effects
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Embryonic Induction / physiology
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Humans
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Inositol Phosphates / metabolism
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Intermediate Filament Proteins / metabolism
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Nerve Tissue Proteins / metabolism
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Nervous System / cytology
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Nervous System / embryology*
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Nestin
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Neurogenesis / drug effects
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Neurogenesis / physiology*
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Neurons / cytology
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Neurons / drug effects
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Neurons / metabolism*
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Phosphopyruvate Hydratase / metabolism
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Receptors, Purinergic / drug effects
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Receptors, Purinergic / metabolism*
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Receptors, Purinergic P2 / drug effects
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Receptors, Purinergic P2 / metabolism
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Receptors, Purinergic P2X2
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Receptors, Purinergic P2Y1
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Receptors, Purinergic P2Y2
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Stem Cells / drug effects
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Stem Cells / metabolism
Substances
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Inositol Phosphates
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Intermediate Filament Proteins
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NES protein, human
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Nerve Tissue Proteins
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Nestin
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P2RX2 protein, human
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P2RY1 protein, human
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P2RY2 protein, human
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Receptors, Purinergic
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Receptors, Purinergic P2
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Receptors, Purinergic P2X2
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Receptors, Purinergic P2Y1
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Receptors, Purinergic P2Y2
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inositol 3,4,5-trisphosphate
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purinoceptor P2Y6
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Adenosine Triphosphate
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Phosphopyruvate Hydratase
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Calcium