title: Modulation of mouse embryonic stem cell proliferation and neural differentiation by the P2X7 receptor.
abstract: BACKGROUND
  Novel developmental functions have been attributed to the P2X7 receptor (P2X7R) including proliferation stimulation and neural differentiation. Mouse embryonic stem cells (ESC), induced with retinoic acid to neural differentiation, closely assemble processes occurring during neuroectodermal development of the early embryo.
  
  
  RESULTS
  P2X7R expression together with the pluripotency marker Oct-4 was highest in undifferentiated ESC. In undifferentiated cells, the P2X7R agonist Bz-ATP accelerated cell cycle entry, which was blocked by the specific P2X7R inhibitor KN-62. ESC induced to neural differentiation with retinoic acid, reduced Oct-4 and P2X7R expression. P2X7R receptor-promoted intracellular calcium fluxes were obtained at lower Bz-ATP ligand concentrations in undifferentiated and in neural-differentiated cells compared to other studies. The presence of KN-62 led to increased number of cells expressing SSEA-1, Dcx and β3-tubulin, as well as the number of SSEA-1 and β3-tubulin-double-positive cells confirming that onset of neuroectodermal differentiation and neuronal fate determination depends on suppression of P2X7R activity. Moreover, an increase in the number of Ki-67 positive cells in conditions of P2X7R inhibition indicates rescue of progenitors into the cell cycle, augmenting the number of neuroblasts and consequently neurogenesis.
  
  
  CONCLUSIONS
  In embryonic cells, P2X7R expression and activity is upregulated, maintaining proliferation, while upon induction to neural differentiation P2X7 receptor expression and activity needs to be suppressed.
authors.affiliation: Departamento de Bioquímica; Instituto de Química, Universidade de São Paulo, São Paulo, Brasil.
authors.forename: Talita
authors.initials: T
authors.lastname: Glaser
authors.affiliation: Departamento de Bioquímica; Instituto de Química, Universidade de São Paulo, São Paulo, Brasil.
authors.forename: Sophia La Banca
authors.initials: SL
authors.lastname: de Oliveira
authors.affiliation: Departamento de Bioquímica; Instituto de Química, Universidade de São Paulo, São Paulo, Brasil.
authors.forename: Arquimedes
authors.initials: A
authors.lastname: Cheffer
authors.affiliation: Departamento de Bioquímica; Instituto de Química, Universidade de São Paulo, São Paulo, Brasil.
authors.forename: Renata
authors.initials: R
authors.lastname: Beco
authors.affiliation: Departamento de Bioquímica; Instituto de Química, Universidade de São Paulo, São Paulo, Brasil.
authors.forename: Patrícia
authors.initials: P
authors.lastname: Martins
authors.affiliation: Departamento de Bioquímica; Instituto de Química, Universidade de São Paulo, São Paulo, Brasil.
authors.forename: Maynara
authors.initials: M
authors.lastname: Fornazari
authors.affiliation: Departamento de Bioquímica; Instituto de Química, Universidade de São Paulo, São Paulo, Brasil.
authors.forename: Claudiana
authors.initials: C
authors.lastname: Lameu
authors.affiliation: Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro - UFRJ, Rio de Janeiro, RJ, Brazil.
authors.forename: Helio Miranda Costa
authors.initials: HM
authors.lastname: Junior
authors.affiliation: Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro - UFRJ, Rio de Janeiro, RJ, Brazil.
authors.forename: Robson
authors.initials: R
authors.lastname: Coutinho-Silva
authors.affiliation: Departamento de Bioquímica; Instituto de Química, Universidade de São Paulo, São Paulo, Brasil.
authors.forename: Henning
authors.initials: H
authors.lastname: Ulrich
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation; D049109:Cell Proliferation; D000089662:Doublecortin Protein; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D055786:Gene Knockout Techniques; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D058486:Receptors, Purinergic P2X7 / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D015854:Up-Regulation
chemical_list: C000714847:Dcx protein, mouse; D000089662:Doublecortin Protein; D050814:Octamer Transcription Factor-3; D058486:Receptors, Purinergic P2X7; D002118:Calcium