title: [Morphologic evaluation and Ca2+ mobilization by glicose and acetylcholine in human pancreatic cells].
abstract: OBJECTIVE
  The proposal of this study was to analyze morphology of the organelles and cytoskeleton in human pancreatic cells cultured and the mobilization of the cytosolic calcium ([Ca2+]c) in response to glucose and ACh by fluorimetry method.
  
  
  METHODS
  The cells were plated on glass coverslips, fixed and stained with a combination of fluorophores: the nuclei were stained with DAPI and mitochondria with Mytotracker Red. It was used phalloidin and the secondary antibodies Alexa Fluor conjugated green and red-fluorescent (488 and 594) to identify the protein cell actin F and type M3 muscarinic receptor respectively. The cells also were loaded with fura-2/AM to study Ca2+ mobilization.
  
  
  RESULTS
  The human pancreatic cells show characteristics morphologically preserved with great amount of mitochondria. In region major cell density was evidenced pseudo-islets and type M3 muscarinic receptors. Through increase of [Ca2+]c due to action of glucose and ACh were shown that the cells capacity to respond to these stimuli were conserved. The elevation of the [Ca2+]c depended on concentration by glucose-induced promoting sustained phase and ACh-induced a biphasic response.
  
  
  CONCLUSIONS
  The morphologic characteristics of human pancreatic cells cultured were preserved. The Ca2+ mobilization in response to glucose and ACh confirmed its functionality. The expression of the M3 muscarinic receptors in human pancreatic cell cultured was demonstrated.
authors.affiliation: Departamento de Biofísica, EPM, UNIFESP, São Paulo, SP. daniela@biofis.epm.br
authors.forename: Daniela Espinha
authors.initials: DE
authors.lastname: Cardoso
authors.forename: Lucimar Pereira de
authors.initials: LP
authors.lastname: França
authors.forename: Elisângela
authors.initials: E
authors.lastname: Chinen
authors.forename: Andréa A F Souza
authors.initials: AA
authors.lastname: Moraes
authors.forename: Alice Teixeira
authors.initials: AT
authors.lastname: Ferreira
authors.forename: Jerônimo Pereira de
authors.initials: JP
authors.lastname: França
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology*; D000704:Analysis of Variance; D020013:Calcium Signaling / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods; D053144:Cell Nucleus Shape; D002478:Cells, Cultured; D018679:Cholinergic Agonists / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D007150:Immunohistochemistry; D007328:Insulin / Q000096:biosynthesis / Q000378:metabolism / Q000502:physiology*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000502:physiology; D007515:Islets of Langerhans / Q000737:chemistry / Q000166:cytology / Q000187:drug effects* / Q000648:ultrastructure; D015388:Organelles / Q000737:chemistry; D043587:Receptor, Muscarinic M3 / Q000737:chemistry / Q000378:metabolism
chemical_list: D018679:Cholinergic Agonists; D007328:Insulin; D043587:Receptor, Muscarinic M3; D005947:Glucose; D000109:Acetylcholine