Cultured Müller cells have high levels of epidermal growth factor receptors.
High levels of epidermal growth factor (EGF)-receptors have been reported in membrane homogenates of bovine retinas, but the biologic function and tissue target of EGF in the retina have not been established fully. Because EGF participation has been suggested in the mechanisms of wound healing and Müller cells undergo changes after retinal injury, the authors studied EGF receptor expression and functional role of this substance in cultured Müller cells. These cells (isolated from normal rats) were tested for the glial cell markers: vimentin, S-100 protein, and carbonic anhydrase C. These markers were found to be positive through all passages used in the experiments. The 125I-EGF binding in Müller cells was highly specific, concentration dependent, and saturable. Compared with 3T3 fibroblasts, Müller cells bound threefold more EGF. Binding kinetics and Scatchard analyses showed the higher level of binding was related to the greater number of receptors on these cells (Müller cells, 2.4 x 10(5) receptors/cell; 3T3 fibroblasts, 7.1 x 10(4) receptors/cell) rather than a change in affinity of the receptors to bind the ligand. Nonlinear-regression analyses suggested the presence of two classes of affinity sites. The high level of EGF-receptor expression in Müller cells was confirmed by western blot analyses that showed increased reactivity of the approximately 170-kilodalton receptor band to a monoclonal anti-EGF receptor antibody. Moreover, EGF treatment of Müller cells resulted in two- to threefold increase in DNA synthesis, as evidenced by 3H-thymidine uptake studies. These findings support a functional role for EGF in Müller cell proliferation in retinal disease.
Roque
R S
RS
Department of Cellular Biology, Medical College of Georgia, Augusta 30912.
Caldwell
R B
RB
Behzadian
M A
MA
D016475:3T3 Cells / Q000378:metabolism; D000818:Animals; D002256:Carbonic Anhydrases / Q000378:metabolism; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism*; D007124:Immunoenzyme Techniques; D051379:Mice; D009457:Neuroglia / Q000378:metabolism; D011869:Radioligand Assay; D051381:Rats; D011922:Rats, Mutant Strains; D012160:Retina / Q000166:cytology / Q000378:metabolism*; D009418:S100 Proteins / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism
D009418:S100 Proteins; D014746:Vimentin; D004815:Epidermal Growth Factor; D004247:DNA; D066246:ErbB Receptors; D002256:Carbonic Anhydrases