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Han-Qing Xie, Ruben Adler; Green Cone Opsin and Rhodopsin Regulation by CNTF and Staurosporine in Cultured Chick Photoreceptors. Invest. Ophthalmol. Vis. Sci. 2000;41(13):4317-4323.
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purpose. To investigate the regulation of visual pigment expression in chick
embryo photoreceptor cells by ciliary neurotrophic factor (CNTF), and
by the protein kinase inhibitor staurosporine.
methods. Embryonic day (ED) 8 chick embryo retinal cells were dissociated and
cultured at low densities for 3 days, either in control medium or in
medium supplemented with CNTF or staurosporine. The cultures were
analyzed by immunocytochemistry with the monoclonal antibody Rho4D2,
which recognizes chicken rhodopsin and green cone pigment, and by
reverse transcription–polymerase chain reaction (RT-PCR) and Northern
blot analysis to investigate visual pigment expression at the mRNA
results. CNTF increased the number of Rho4D2-immunoreactive photoreceptors in
retinal cell cultures, in agreement with previous reports. RT-PCR and
Northern blot analysis, however, showed that rhodopsin mRNA was
undetectable in both control and CNTF-treated cultures but that CNTF
induced significant increases in mRNA levels for the green cone
pigment. Staurosporine-treated cultures also had more
Rho4D2-immunoreactive cells than control cultures, but this increase
was accompanied by induction of rhodopsin expression, with concomitant
decreases in levels of green cone pigment mRNA. No significant
differences were found between CNTF- or staurosporine-treated cultures
and the corresponding control cultures regarding the red cone pigment,
which was expressed in all cases, and the blue and violet pigments,
which were not detected in any of the samples.
conclusions. The results suggest that multiple regulatory systems control visual
pigment expression during differentiation of chick embryo photoreceptor
cells. CNTF appears to stimulate specifically the differentiation of
green cones, without the previously suggested effects on the
differentiation of rod photoreceptors in ED 8 chick retinal
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