Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2025 Jan 9;26(2):496. doi: 10.3390/ijms26020496

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2025 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Developmental anomalies of the retina and eyeball caused by local misexpression of exogenous Shh. A DNA solution containing the Shh and GFP expression plasmids was electroporated in a restricted area around the right eye primordium of HH stage 14 chick embryos, and the resulting morphology was examined at stage 26. (A) Views of the affected eye and its fluorescent image show that the injected constructs are expressed in the upper anterior portion of the right eye. The growth of the upper anterior half of the eyeball is retarded and the cornea and lens have developed in an upward anterior direction (arrowhead). Ten times magnification by stereo microscope. (B) Views of the affected eye and its fluorescent image matched with the upper panel show that the injected constructs are expressed in the lower part of the right eye. The growth of the lower half of the eyeball is retarded and the cornea and lens developed in a downward direction (arrowhead). (C) The coronal section of the eyes shows that the growth of the bottom half of the right eyeball is retarded (inset a), and the cornea and lens are at the bottom corner (HE staining, bar scale 60 μm). (D) Immunohistochemistry with specific antibodies reveals exogenous Shh and GFP protein (asterisk) in the treated area of the Shh-induced right eye (inset a in panel C) but not in the uninduced left eye (inset b in panel C). There is less Pax6 expression in the treated area of the Shh-induced eye compared to the same area of the uninduced eye (bar scale 30 μm). (E) Immunohistochemistry with an anti-BrdU antibody shows slightly weak staining in the Shh-induced area (inset c in panel D) compared to the uninduced area (inset d in panel D). Apoptosis is slightly detected in the retina and pigment epithelia (arrowheads) in the Shh-induced and uninduced eyes, suggesting that gene induction did not result in cell death. Immunohistochemistry and in situ hybridization suggest decreased expression of Pax6, Six3 and Rx in the Shh-induced right eye (inset a in panel C) compared to the uninduced left eye (inset b in panel C), but mostly the same level of Musashi expression in both eyes (bar scale 20 μm). (F) qRT-PCR analysis suggests that expression of Gli1, Pax6, Six3 and Rx decreased in the affected right eye (dissected area of inset c in panel D) compared to the unaffected left eye (dissected area of inset d in panel D), but that there was mostly the same level of Musashi expression in both eyes. Expression of Patched, Gli1, Pax6, Musashi, Six3 and Rx in the Shh-induced and uninduced retinas is indicated by qRT-PCR. The amount of the cDNA pools was prepared for qRT-PCR. The bar graph indicates the ratio of Shh-induced to Shh-uninduced retinas. The analysis is representative of three independent experiments. *, p < 0.05 (unpaired t-test, two-tailed); error bars, ±SD.