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Fig. 6. HtrA1 antagonizes Bmp signaling in vivo. Cultured 293T cells expressing
HtrA1 (A-I) or noggin (J-R) were injected near the right eyes of stage 9-10
chick embryos, or the HtrA1 expression vector was electroporated into
the right optic cups of stage 10-11 chick embryos (S,T,U). After incubation
until stage 23-24 (A-R) or stage 17-18 (S,T,U), embryos were harvested. (A,J)
Localization of injected cells as monitored by GFP fluorescence. Pictures of
the eyes were taken at a constant magnification and image-analyzed for size.
The lens or retina was judged to be small when the area of the lens or the
pigmented retina in the picture was less than 70% of the untreated side. (B,K)
Eyes in un-injected sides. (C,L) Eyes injected with HtrA1- or
noggin-expressing cells. (D,M) Frontal sections of control eyes. (E,N) Frontal
sections of injected eyes. (F,H,O,Q) Magnified views of D and M. (G,I,P,R)
Magnified views of E and N, showing abnormalities of the lens and ventral
retina caused by HtrA1 and noggin, respectively. Note that the retinal pigment
epithelium (regions between arrowheads in D and M) was replaced by a
neuroepithelium-like tissue (regions between arrowheads in E and N). Vertical
lines in F, G, O and P indicate the thickness of the primary lens fibers. (S)
Retina showing expression of co-electroporated GFP. (T) Vax
expression in the untreated eye. (U) Upregulated Vax expression in
the retina caused by overexpression of HtrA1. Eyes tended to be
smaller after electroporation with the HtrA1 expression vector. Upregulation
of Vax expression was seen in eyes of normal or slightly small
size.
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