Constancy of ERp29 expression in cultured retinal pigment epithelial cells in the Ccl2/Cx3cr1 deficient mouse model of age-related macular degeneration.
OBJECTIVE
Given the critical role of the retinal pigment epithelium (RPE) in the pathogenesis of age-related macular generation (AMD) and the links drawn between chaperone proteins and neurodegenerative disease, we aimed to culture RPE from the Ccl2(-/-)/Cx3cr1(-/-) mouse model of AMD and evaluate expression of chaperone protein ERp29.


METHODS
Murine RPE cells were surgically and chemically isolated and cultured. ERp29 mRNA and protein expression were evaluated by real-time RT-PCR, immunohistochemistry, and Western blot.


RESULTS
Ccl2(-/-)/Cx3cr1(-/-) RPE was successfully isolated and cultured. They presented a decreased but statistically insignificant difference in ERp29 transcript and protein expression compared to C57B6/L wild type mouse RPE.


CONCLUSIONS
The effective murine RPE culture described here enables future investigation into RPE biology and AMD pathogenesis. Although we found a decrease in ERp29 expression in the Ccl2(-/-)/Cx3cr1(-/-) RPE, the difference was less than we previously observed in the whole retina. This suggests that the RPE may not contribute to the greater differential expression and ERp29 might have a more significant role in the neuroretina than in the RPE during AMD pathogenesis.
Verma
Varun
V
Immunopathology Section, Laboratory of Immunology, National Eye Institute, National Institutes of Health, Bethesda, Maryland 20892-1857, USA.
Sauer
Theodor
T
Chan
Chi-Chao
CC
Zhou
Min
M
Zhang
Congxiao
C
Maminishkis
Arvydas
A
Shen
Defen
D
Tuo
Jingsheng
J
D000818:Animals; D000073883:CX3C Chemokine Receptor 1; D002469:Cell Separation; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000172:deficiency / Q000235:genetics*; D004195:Disease Models, Animal; D015870:Gene Expression; D006360:Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D007124:Immunoenzyme Techniques; D008268:Macular Degeneration / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010857:Pigment Epithelium of Eye / Q000378:metabolism* / Q000648:ultrastructure; D012333:RNA, Messenger / Q000378:metabolism; D019707:Receptors, Chemokine / Q000172:deficiency / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000073883:CX3C Chemokine Receptor 1; C491768:Ccl2 protein, mouse; D018932:Chemokine CCL2; C471885:Cx3cr1 protein, mouse; C491737:Erp29 protein, mouse; D006360:Heat-Shock Proteins; D012333:RNA, Messenger; D019707:Receptors, Chemokine