PMID- 21409522
OWN - NLM
STAT- MEDLINE
DCOM- 20111130
LR  - 20211203
IS  - 1573-6830 (Electronic)
IS  - 0272-4340 (Linking)
VI  - 31
IP  - 6
DP  - 2011 Aug
TI  - Functional identification of cell phenotypes differentiating from mice retinal 
      neurospheres using single cell calcium imaging.
PG  - 835-46
LID - 10.1007/s10571-011-9673-6 [doi]
AB  - Degeneration of neural retina causes vision impairment and can lead to blindness. 
      Neural stem and progenitor cells might be used as a tool directed to regenerative 
      medicine of the retina. Here, we describe a novel platform for cell 
      phenotype-specific drug discovery and screening of proneurogenic factors, able to 
      boost differentiation of neural retinal progenitor cells. By using single cell 
      calcium imaging (SCCI) and a rational-based stimulation protocol, a diversity of 
      cells emerging from differentiated retinal neurosphere cultures were identified. 
      Exposure of retinal progenitor cultures to KCl or to 
      alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA) stimulated Ca(2+) 
      transients in microtubule-associated protein 2 (MAP-2) positive neurons. 
      Doublecortin (DCX) and polysialated neural cell adhesion molecule (PSA-NCAM) 
      positive neuroblasts were distinguished from differentiated neurons on the basis 
      of their response to muscimol. Ca(2+) fluxes in glial fibrillary acidic protein 
      (GFAP) or glutamine synthetase (GS) positive cells were induced by ATP. To 
      validate the platform, neurospheres were treated with brain-derived neurotrophic 
      factor (BDNF) (proneurogenic) or ciliary neurotrophic factor (CNTF) (gliogenic 
      factor). BDNF increased the percentage of differentiated cells expressing Tuj-1 
      sensitive to KCl or AMPA and reduced the population of cells responding to 
      muscimol. CNTF exposure resulted in a higher number of cells expressing GFAP 
      responding to ATP. All together, our data may open new perspectives for cell 
      type-specific discovery of drug targets and screening of novel proneurogenic 
      factors to boost differentiation of neural retina cells to treat degenerative 
      retinal diseases.
FAU - De Melo Reis, R A
AU  - De Melo Reis RA
AD  - Neuroprotection and Neurogenesis in Brain Repair, Center for Neurosciences and 
      Cell Biology of Coimbra, Faculty of Medicine, University of Coimbra, 3004-504 
      Coimbra, Portugal.
FAU - Schitine, C S
AU  - Schitine CS
FAU - Kofalvi, A
AU  - Kofalvi A
FAU - Grade, S
AU  - Grade S
FAU - Cortes, L
AU  - Cortes L
FAU - Gardino, P F
AU  - Gardino PF
FAU - Malva, J O
AU  - Malva JO
FAU - de Mello, F G
AU  - de Mello FG
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110317
PL  - United States
TA  - Cell Mol Neurobiol
JT  - Cellular and molecular neurobiology
JID - 8200709
RN  - 0 (Biomarkers)
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Ciliary Neurotrophic Factor)
RN  - 0 (Dcx protein, mouse)
RN  - 0 (Doublecortin Protein)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Animals
MH  - Biomarkers/metabolism
MH  - Brain-Derived Neurotrophic Factor/pharmacology
MH  - Calcium/*metabolism
MH  - *Cell Differentiation/drug effects
MH  - Cell Lineage/drug effects
MH  - Ciliary Neurotrophic Factor/pharmacology
MH  - Doublecortin Protein
MH  - Imaging, Three-Dimensional/*methods
MH  - Mice
MH  - Neuroglia/cytology/drug effects/metabolism
MH  - Neurons/*cytology/drug effects/metabolism
MH  - Phenotype
MH  - Retina/*cytology
MH  - Single-Cell Analysis/*methods
MH  - Spheroids, Cellular/*cytology/drug effects/metabolism
MH  - Stem Cells/cytology/drug effects/metabolism
EDAT- 2011/03/17 06:00
MHDA- 2011/12/13 00:00
CRDT- 2011/03/17 06:00
PHST- 2010/12/21 00:00 [received]
PHST- 2011/03/02 00:00 [accepted]
PHST- 2011/03/17 06:00 [entrez]
PHST- 2011/03/17 06:00 [pubmed]
PHST- 2011/12/13 00:00 [medline]
AID - 10.1007/s10571-011-9673-6 [doi]
PST - ppublish
SO  - Cell Mol Neurobiol. 2011 Aug;31(6):835-46. doi: 10.1007/s10571-011-9673-6. Epub 
      2011 Mar 17.