title: Differentiated human midbrain-derived neural progenitor cells express excitatory strychnine-sensitive glycine receptors containing α2β subunits.
abstract: BACKGROUND
  Human fetal midbrain-derived neural progenitor cells (NPCs) may deliver a tissue source for drug screening and regenerative cell therapy to treat Parkinson's disease. While glutamate and GABA(A) receptors play an important role in neurogenesis, the involvement of glycine receptors during human neurogenesis and dopaminergic differentiation as well as their molecular and functional characteristics in NPCs are largely unknown.
  
  
  RESULTS
  Here we investigated NPCs in respect to their glycine receptor function and subunit expression using electrophysiology, calcium imaging, immunocytochemistry, and quantitative real-time PCR. Whole-cell recordings demonstrate the ability of NPCs to express functional strychnine-sensitive glycine receptors after differentiation for 3 weeks in vitro. Pharmacological and molecular analyses indicate a predominance of glycine receptor heteromers containing α2β subunits. Intracellular calcium measurements of differentiated NPCs suggest that glycine evokes depolarisations mediated by strychnine-sensitive glycine receptors and not by D-serine-sensitive excitatory glycine receptors. Culturing NPCs with additional glycine, the glycine-receptor antagonist strychnine, or the Na(+)-K(+)-Cl(-) co-transporter 1 (NKCC1)-inhibitor bumetanide did not significantly influence cell proliferation and differentiation in vitro.
  
  
  CONCLUSIONS
  These data indicate that NPCs derived from human fetal midbrain tissue acquire essential glycine receptor properties during neuronal maturation. However, glycine receptors seem to have a limited functional impact on neurogenesis and dopaminergic differentiation of NPCs in vitro.
authors.affiliation: Department of Neurology, Hannover Medical School, Hannover, Germany. wegner.florian@mh-hannover.de
authors.forename: Florian
authors.initials: F
authors.lastname: Wegner
authors.forename: Robert
authors.initials: R
authors.lastname: Kraft
authors.forename: Kathy
authors.initials: K
authors.lastname: Busse
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Härtig
authors.forename: Jörg
authors.initials: J
authors.lastname: Ahrens
authors.forename: Andreas
authors.initials: A
authors.lastname: Leffler
authors.forename: Reinhard
authors.initials: R
authors.lastname: Dengler
authors.forename: Johannes
authors.initials: J
authors.lastname: Schwarz
mesh_terms: D001483:Base Sequence; D020013:Calcium Signaling; D002454:Cell Differentiation; D002478:Cells, Cultured; D017931:DNA Primers / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008636:Mesencephalon / Q000166:cytology / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D060888:Real-Time Polymerase Chain Reaction; D018009:Receptors, Glycine / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013331:Strychnine / Q000494:pharmacology
chemical_list: D017931:DNA Primers; C439504:GLRA2 protein, human; C114561:GLRB protein, human; D018009:Receptors, Glycine; D013331:Strychnine