Suppression of lens growth by alphaA-crystallin promoter-driven expression of diphtheria toxin results in disruption of retinal cell organization in zebrafish.
In order to study lens-retina relationships during development, we cloned the zebrafish alphaA-crystallin cDNA and its promoter region. Using a 2.8-kb fragment of the zebrafish alphaA-crystallin promoter (z(alpha)Acry), we expressed the diphtheria toxin A fragment (DTA) in zebrafish embryos in a lens-specific manner. Injection of the z(alpha)Acry-DTA plasmid into eggs at the one-or two-cell stage resulted in the formation of small eyes, in which both lens and retina were reduced in size. In the DTA-expressing lenses, their fiber structure was disorganized, indicating that normal lens development had been abrogated. The neural retina also showed abnormal development, although this tissue did not express DTA. Lamination in the retina did not develop well, and molecular markers for the outer and inner plexiform layers were either abnormally expressed or absent. However, cell type-specific markers of ganglion and bipolar cells, as well as photoreceptors, were expressed in appropriate positions, indicating that initial differentiation of these retinal subpopulations occurred in the DTA-expressing embryos. Cell proliferation also proceeded normally in these embryos, although apoptosis was enhanced. These results suggest that the differentiated lens plays a critical role in the morphogenetic organization of retinal cells during eye development in zebrafish embryos.
Kurita
Ryo
R
Department of Molecular and Developmental Biology, Institute of Medical Science, University of Tokyo, Japan.
Sagara
Hiroshi
H
Aoki
Yutaka
Y
Link
Brian A
BA
Arai
Ken-ichi
K
Watanabe
Sumiko
S
D000818:Animals; D017209:Apoptosis; D001483:Base Sequence; D002454:Cell Differentiation; D002455:Cell Division; D004167:Diphtheria Toxin / Q000235:genetics*; D004625:Embryo, Nonmammalian / Q000648:ultrastructure; D017353:Gene Deletion; D015870:Gene Expression; D007908:Lens, Crystalline / Q000196:embryology*; D009154:Mutation; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D012160:Retina / Q000166:cytology / Q000196:embryology* / Q000648:ultrastructure; D016415:Sequence Alignment; D019076:Transgenes; D015027:Zebrafish; D038201:alpha-Crystallins / Q000235:genetics* / Q000378:metabolism
D004167:Diphtheria Toxin; D012333:RNA, Messenger; D038201:alpha-Crystallins