title: Pharmacological properties of purinergic receptors and their effects on proliferation and induction of neuronal differentiation of P19 embryonal carcinoma cells.
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.
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Av. Prof. Lineu Prestes 748, 05508-900 São Paulo, SP, Brazil.
authors.forename: R R
authors.initials: RR
authors.lastname: Resende
authors.forename: L R G
authors.initials: LR
authors.lastname: Britto
authors.forename: H
authors.initials: H
authors.lastname: Ulrich
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000502:physiology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D054278:Embryonal Carcinoma Stem Cells; D004627:Embryonic Induction / Q000187:drug effects / Q000502:physiology; D006801:Humans; D007295:Inositol Phosphates / Q000378:metabolism; D007381:Intermediate Filament Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009420:Nervous System / Q000166:cytology / Q000196:embryology*; D064231:Nestin; D055495:Neurogenesis / Q000187:drug effects / Q000502:physiology*; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D011983:Receptors, Purinergic / Q000187:drug effects / Q000378:metabolism*; D018048:Receptors, Purinergic P2 / Q000187:drug effects / Q000378:metabolism; D058476:Receptors, Purinergic P2X2; D058472:Receptors, Purinergic P2Y1; D058473:Receptors, Purinergic P2Y2; D013234:Stem Cells / Q000187:drug effects / Q000378:metabolism
chemical_list: D007295:Inositol Phosphates; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; C546401:P2RX2 protein, human; C546382:P2RY1 protein, human; C546385:P2RY2 protein, human; D011983:Receptors, Purinergic; D018048:Receptors, Purinergic P2; D058476:Receptors, Purinergic P2X2; D058472:Receptors, Purinergic P2Y1; D058473:Receptors, Purinergic P2Y2; C118500:inositol 3,4,5-trisphosphate; C099845:purinoceptor P2Y6; D000255:Adenosine Triphosphate; D010751:Phosphopyruvate Hydratase; D002118:Calcium