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Journal of Neuroscience, Vol 12, 2022-2033, Copyright © 1992 by Society for Neuroscience
Fibroblast growth factor stimulates photoreceptor differentiation in vitro
D Hicks and Y Courtois
INSERM U. 118, Paris, France.
Dissociated newborn rat retinal cells were maintained in monolayer culture
for periods of up to 11 d. When grown in the absence of exogenous growth
factors, 1-2% of the total neuronal population expressed opsin (the
photopigment that is specific for maturing photoreceptors). Addition of a
single dose of 10 ng/ml basic fibroblast growth factor (bFGF) to the
culture medium induced an average increase of sixfold in the numbers of
neurons expressing opsin. This supplementation had little effect on the
total number of differentiated neurons or of glial cells when measured at
the same time points. Furthermore, another specific class of retinal
neurons, the amacrine cells, showed no changes following exposure to this
growth factor. Two other growth factors known to exert neurotrophic
effects, epidermal and nerve growth factor, were without effect. The effect
of bFGF was dose dependent, with highly significant differences being
observed with as little as 100 pg/ml, and with 700 pg/ml eliciting
half-maximal stimulation; maximal effects were observed at 10 ng/ml.
Induction of opsin expression by low concentrations of bFGF was blocked
completely by an antiserum directed specifically against bFGF, but not by
preimmune serum immunoglobulins. This increase in the number of
photoreceptors expressing opsin following exposure to bFGF could have been
due to either increased cell survival, increased proliferation of
progenitor cells, or increased differentiation of immature photoreceptors.
There was no increase in overall cell survival under the experimental
conditions used, and double labeling immunocytochemistry combined with
autoradiographic analysis of 3H- thymidine uptake showed that proliferation
of neuronal precursors was not enhanced by the addition of bFGF. In
contrast to these observations, cultures established from older (postnatal
day 3) retina revealed large numbers of opsin-expressing photoreceptors in
all culture plates, with or without added growth factors. This reduction in
the stimulatory effects of bFGF with increasing postnatal age is consistent
with the period of sensitivity being limited to the cycling of neuronal
precursors. It is possible that a bFGF-like molecule is secreted by
neighboring cells such as the retinal pigmented epithelium, to participate
in retinal development and differentiation. To our understanding, this
molecule is the first protein identified to influence specifically the
differentiation of photoreceptor cells.
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