PMID- 9660494
OWN - NLM
STAT- MEDLINE
DCOM- 19980714
LR  - 20071114
IS  - 0146-0404 (Print)
IS  - 0146-0404 (Linking)
VI  - 39
IP  - 8
DP  - 1998 Jul
TI  - Antigenic changes of rabbit retinal Muller cells in culture.
PG  - 1453-61
AB  - PURPOSE: To determine whether dissociated and cultured Muller cells from the 
      avascular rabbit retina undergo the same phenotypic changes as Muller cells that 
      are dissociated and cultured from a vascular retina. METHODS: Muller cells were 
      dissociated from adult rabbit retinas by using an enzymatic digestion-mechanical 
      trituration technique and a cell attachment method that provided Muller cell- 
      enriched cell cultures. Indirect immunofluorescence localization of vimentin, 
      glial fibrillary acidic protein (GFAP), glutamine synthetase (GS), beta-amyloid 
      precursor protein (beta-APP), and (alpha-smooth muscle actin (alpha-SMA) was 
      carried out on Muller cells that were freshly dissociated, on those that had been 
      in culture 2 and 6 days, and on confluent primary cultures and late-passage 
      cultures. The specificity of the antibodies and changes in protein expression 
      were examined by western blot analysis. RESULTS: The expression of vimentin, 
      GFAP, GS, and beta-APP was present 2 days after dissociation and was retained 
      through 6 days in culture, at which time alpha-SMA began to be expressed in a 
      small number of cells. The confluent, primary cultures no longer expressed GS, 
      but vimentin and beta-APP were still expressed, and the expression of alpha-SMA 
      was increased. During the late-passage stage, the morphologic appearance of the 
      Muller cell cultures was large and amorphous, with additional changes in 
      antigenicity. Although there was loss of expression of the intermediate filament 
      proteins GFAP and vimentin, the expression of beta-APP was maintained, whereas 
      alpha-SMA was increased and appeared to be a major cytoskeletal protein. 
      CONCLUSIONS: Dissociated Muller cells that were maintained in culture underwent 
      phenotypic changes that included a large, amorphous appearance; the loss of 
      detectable vimentin, GFAP, and GS expression; the persistent presence of 
      beta-APP; and the de novo appearance of alpha-SMA. The phenotypic and antigenic 
      changes that occurred in cultured Muller cells from an avascular retina were 
      similar but not identical to the changes observed in cultured Muller cells from a 
      vascular retina.
FAU - McGillem, G S
AU  - McGillem GS
AD  - Eye Foundation Hospital, Department of Ophthalmology, University of Alabama at 
      Birmingham, 35233, USA.
FAU - Guidry, C
AU  - Guidry C
FAU - Dacheux, R F
AU  - Dacheux RF
LA  - eng
GR  - EY-03011/EY/NEI NIH HHS/United States
GR  - EY-09536/EY/NEI NIH HHS/United States
GR  - P30EY03039/EY/NEI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Invest Ophthalmol Vis Sci
JT  - Investigative ophthalmology & visual science
JID - 7703701
RN  - 0 (Actins)
RN  - 0 (Amidines)
RN  - 0 (Amyloid beta-Protein Precursor)
RN  - 0 (Epitopes)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Glial Fibrillary Acidic Protein)
RN  - 0 (Vimentin)
RN  - 88746-72-9 (diamidino yellow)
RN  - EC 6.3.1.2 (Glutamate-Ammonia Ligase)
SB  - IM
MH  - Actins/metabolism
MH  - Amidines
MH  - Amyloid beta-Protein Precursor/metabolism
MH  - Animals
MH  - Blotting, Western
MH  - Cells, Cultured
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Epitopes/*immunology
MH  - Fluorescent Antibody Technique, Indirect
MH  - Fluorescent Dyes
MH  - Glial Fibrillary Acidic Protein/metabolism
MH  - Glutamate-Ammonia Ligase/metabolism
MH  - Neuroglia/*immunology/metabolism
MH  - Phenotype
MH  - Rabbits
MH  - Retina/*immunology/metabolism
MH  - Vimentin/metabolism
EDAT- 1998/07/11 00:00
MHDA- 1998/07/11 00:01
CRDT- 1998/07/11 00:00
PHST- 1998/07/11 00:00 [pubmed]
PHST- 1998/07/11 00:01 [medline]
PHST- 1998/07/11 00:00 [entrez]
PST - ppublish
SO  - Invest Ophthalmol Vis Sci. 1998 Jul;39(8):1453-61.