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Volume 17, Number 16,
Issue of August 15, 1997
pp. 6277-6288
Copyright ©1997 Society for Neuroscience
Sonic Hedgehog Promotes Rod Photoreceptor Differentiation in
Mammalian Retinal Cells In Vitro
Received April 21, 1997; revised May 20, 1997; accepted May 27, 1997.
Edward M. Levine,
Henk Roelink,
Jennifer Turner, and
Thomas A. Reh
Department of Biological Structure, University of Washington,
Seattle, Washington 98195
The hedgehog gene family encodes secreted proteins
important in many developmental patterning events in both vertebrates
and invertebrates. In the Drosophila eye disk, hedgehog
controls the progression of photoreceptor differentiation in the
morphogenetic furrow. To investigate whether hedgehog proteins are also
involved in the development of the vertebrate retina at stages of
photoreceptor differentiation, we analyzed expression of the three
known vertebrate hedgehog genes. We found that Sonic
hedgehog and Desert hedgehog are expressed in
the developing retina, albeit at very low levels, whereas Indian
hedgehog (Ihh) is expressed in the developing
and mature retinal pigmented epithelium, beginning at embryonic day 13. To determine whether hedgehog proteins have activities on developing
retinal cells, we used an in vitro system in which much
of retinal histogenesis is recapitulated. N-terminal recombinant Sonic
Hedgehog protein (SHH-N) was added to rat retinal cultures for 3-12 d,
and the numbers of retinal cells of various phenotypes were analyzed by
immunohistochemistry. We found that SHH-N caused a transient increase
in the number of retinal progenitor cells, and a 2- to 10-fold increase
in the number of photoreceptors differentiating in the cultures when
analyzed with three different photoreceptor-specific antigens. In
contrast, the numbers of retinal ganglion cells and amacrine cells were
similar to those in control cultures. These results show that Hedgehog
proteins can regulate mitogenesis and photoreceptor differentiation in
the vertebrate retina, and Ihh is a candidate factor
from the pigmented epithelium to promote retinal progenitor
proliferation and photoreceptor differentiation.
Key words:
Sonic hedgehog;
Indian hedgehog;
Desert hedgehog;
retina;
mitogenesis;
differentiation;
rod photoreceptor
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