PMID- 35377941
OWN - NLM
STAT- MEDLINE
DCOM- 20220406
LR  - 20220614
IS  - 2164-2591 (Electronic)
IS  - 2164-2591 (Linking)
VI  - 11
IP  - 4
DP  - 2022 Apr 1
TI  - Derivation and Characterization of Murine and Amphibian Muller Glia Cell Lines.
PG  - 4
LID - 10.1167/tvst.11.4.4 [doi]
LID - 4
AB  - PURPOSE: Muller glia (MG) in the retina of Xenopus laevis (African clawed frog) 
      reprogram to a transiently amplifying retinal progenitor state after an injury. 
      These progenitors then give rise to new retinal neurons. In contrast, mammalian 
      MG have a restricted neurogenic capacity and undergo reactive gliosis after 
      injury. This study sought to establish MG cell lines from the 
      regeneration-competent frog and the regeneration-deficient mouse. METHODS: MG 
      were isolated from postnatal day 5 GLAST-CreERT; Rbfl/fl mice and from adult (3-5 
      years post-metamorphic) X laevis. Serial adherent subculture resulted in 
      spontaneously immortalized cells and the establishment of two MG cell lines: 
      murine retinal glia 17 (RG17) and Xenopus glia 69 (XG69). They were characterized 
      for MG gene and protein expression by qPCR, immunostaining, and Western blot. 
      Purinergic signaling was assessed with calcium imaging. Pharmacological 
      perturbations with 2'-3'-O-(4-benzoylbenzoyl) adenosine 5'-triphosphate (BzATP) 
      and KN-62 were performed on RG17 cells. RESULTS: RG17 and XG69 cells express 
      several MG markers and retain purinergic signaling. Pharmacological perturbations 
      of intracellular calcium responses with BzATP and KN-62 suggest that the 
      ionotropic purinergic receptor P2X7 is present and functional in RG17 cells. 
      Stimulation of XG69 cells with adenosine triphosphate-induced calcium responses 
      in a dose-dependent manner. CONCLUSIONS: We report the characterization of RG17 
      and XG69, two novel MG cell lines from species with significantly disparate 
      retinal regenerative capabilities. TRANSLATIONAL RELEVANCE: RG17 and XG69 cell 
      line models will aid comparative studies between species endowed with varied 
      regenerative capacity and will facilitate the development of new cell-based 
      strategies for treating retinal degenerative diseases.
FAU - Gallo, Ryan A
AU  - Gallo RA
AD  - Dr. Nasser Ibrahim Al-Rashid Orbital Vision Research Center, Bascom Palmer Eye 
      Institute, Department of Ophthalmology, University of Miami Miller School of 
      Medicine, Miami, FL, USA.
AD  - Medical Scientist Training Program, University of Miami Miller School of 
      Medicine, Miami, FL, USA.
FAU - Qureshi, Farhan
AU  - Qureshi F
AD  - Medical Scientist Training Program, University of Miami Miller School of 
      Medicine, Miami, FL, USA.
FAU - Strong, Thomas A
AU  - Strong TA
AD  - Dr. Nasser Ibrahim Al-Rashid Orbital Vision Research Center, Bascom Palmer Eye 
      Institute, Department of Ophthalmology, University of Miami Miller School of 
      Medicine, Miami, FL, USA.
FAU - Lang, Steven H
AU  - Lang SH
AD  - Dr. Nasser Ibrahim Al-Rashid Orbital Vision Research Center, Bascom Palmer Eye 
      Institute, Department of Ophthalmology, University of Miami Miller School of 
      Medicine, Miami, FL, USA.
FAU - Pino, Kevin A
AU  - Pino KA
AD  - Dr. Nasser Ibrahim Al-Rashid Orbital Vision Research Center, Bascom Palmer Eye 
      Institute, Department of Ophthalmology, University of Miami Miller School of 
      Medicine, Miami, FL, USA.
FAU - Dvoriantchikova, Galina
AU  - Dvoriantchikova G
AD  - Dr. Nasser Ibrahim Al-Rashid Orbital Vision Research Center, Bascom Palmer Eye 
      Institute, Department of Ophthalmology, University of Miami Miller School of 
      Medicine, Miami, FL, USA.
FAU - Pelaez, Daniel
AU  - Pelaez D
AD  - Dr. Nasser Ibrahim Al-Rashid Orbital Vision Research Center, Bascom Palmer Eye 
      Institute, Department of Ophthalmology, University of Miami Miller School of 
      Medicine, Miami, FL, USA.
AD  - Department of Cell Biology, University of Miami Miller School of Medicine, Miami, 
      FL, USA.
LA  - eng
GR  - P30 EY014801/EY/NEI NIH HHS/United States
GR  - T32 GM112601/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Transl Vis Sci Technol
JT  - Translational vision science & technology
JID - 101595919
SB  - IM
MH  - Animals
MH  - *Ependymoglial Cells/metabolism
MH  - Mammals
MH  - Mice
MH  - Neuroglia/metabolism
MH  - Retina
MH  - *Retinal Neurons
MH  - Xenopus laevis
PMC - PMC8994200
COIS- Disclosure: R.A. Gallo, None; F. Qureshi, None; T.A. Strong, None; S.H. Lang, 
      None; K.A. Pino, None; G. Dvoriantchikova, None; D. Pelaez, None
EDAT- 2022/04/05 06:00
MHDA- 2022/04/07 06:00
PMCR- 2022/04/04
CRDT- 2022/04/04 17:11
PHST- 2022/04/04 17:11 [entrez]
PHST- 2022/04/05 06:00 [pubmed]
PHST- 2022/04/07 06:00 [medline]
PHST- 2022/04/04 00:00 [pmc-release]
AID - 2778724 [pii]
AID - TVST-21-4100 [pii]
AID - 10.1167/tvst.11.4.4 [doi]
PST - ppublish
SO  - Transl Vis Sci Technol. 2022 Apr 1;11(4):4. doi: 10.1167/tvst.11.4.4.