Dataset Information

A conserved role of ?A-crystallin in the development of the zebrafish embryonic lens.

ABSTRACT: ?A- and ?B-crystallins are small heat shock proteins that bind thermodynamically destabilized proteins thereby inhibiting their aggregation. Highly expressed in the mammalian lens, the ?-crystallins have been postulated to play a critical role in the maintenance of lens optical properties by sequestering age-damaged proteins prone to aggregation as well as through a multitude of roles in lens epithelial cells. Here, we have examined the role of ?-crystallins in the development of the vertebrate zebrafish lens. For this purpose, we have carried out morpholino-mediated knockdown of ?A-, ?Ba- and ?Bb-crystallin and characterized the gross morphology of the lens. We observed lens abnormalities, including increased reflectance intensity, as a consequence of the interference with expression of these proteins. These abnormalities were less frequent in transgenic zebrafish embryos expressing rat ?A-crystallin suggesting a specific role of ?-crystallins in embryonic lens development. To extend and confirm these findings, we generated an ?A-crystallin knockout zebrafish line. A more consistent and severe lens phenotype was evident in maternal/zygotic ?A-crystallin mutants compared to those observed by morpholino knockdown. The penetrance of the lens phenotype was reduced by transgenic expression of rat ?A-crystallin and its severity was attenuated by maternal ?A-crystallin expression. These findings demonstrate that the role of ?-crystallins in lens development is conserved from mammals to zebrafish and set the stage for using the embryonic lens as a model system to test mechanistic aspects of ?-crystallin chaperone activity and to develop strategies to fine-tune protein-protein interactions in aging and cataracts.

SUBMITTER: Zou P

PROVIDER: S-EPMC4638411 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature

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A conserved role of αA-crystallin in the development of the zebrafish embryonic lens.

Zou Ping P Wu Shu-Yu SY Koteiche Hanane A HA Mishra Sanjay S Levic Daniel S DS Knapik Ela E Chen Wenbiao W Mchaourab Hassane S HS

Experimental eye research 20150704

αA- and αB-crystallins are small heat shock proteins that bind thermodynamically destabilized proteins thereby inhibiting their aggregation. Highly expressed in the mammalian lens, the α-crystallins have been postulated to play a critical role in the maintenance of lens optical properties by sequestering age-damaged proteins prone to aggregation as well as through a multitude of roles in lens epithelial cells. Here, we have examined the role of α-crystallins in the development of the vertebrate ...[more]

PMID: 26149094

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Project description:Heat shock proteins (Hsps) were originally identified as proteins expressed after exposure of cells to environmental stress. Several Hsps were subsequently shown to play roles as molecular chaperones in normal intracellular protein folding and targeting events and to be expressed during discrete periods in the development of several embryonic tissues. However, only recently have studies begun to address the specific developmental consequences of inhibiting Hsp expression to determine whether these molecular chaperones are required for specific developmental events. We have previously shown that the heat-inducible zebrafish hsp70 gene is expressed during a distinct temporal window of embryonic lens formation at normal growth temperatures. In addition, a 1.5-kb fragment of the zebrafish hsp70 gene promoter is sufficient to direct expression of a gfp reporter gene to the lens, suggesting that the hsp70 gene is expressed as part of the normal lens development program. Here, we used microinjection of morpholino-modified antisense oligonucleotides (MOs) to reduce Hsp70 levels during zebrafish development and to show that Hsp70 is required for normal lens formation. Hsp70-MO-injected embryos exhibited a small-eye phenotype relative to wild-type and control-injected animals, with the phenotype discernable during the second day of development. Histological and immunological analysis revealed a small, underdeveloped lens. Numerous terminal deoxynucleotidyl transferase-mediated dUTP-fluoroscein nick-end labeling (TUNEL)-positive nuclei appeared in the lens of small-eye embryos after 48 hours postfertilization (hpf), whereas they were no longer apparent in untreated embryos by this age. Lenses transplanted from hsp70-MO-injected embryos into wild-type hosts failed to recover and retained the immature morphology characteristic of the small-eye phenotype, indicating that the lens phenotype is lens autonomous. Our data suggest that the lens defect in hsp70-MO-injected embryos is predominantly at the level of postmitotic lens fiber differentiation, a result supported by the appearance of mature lens organization in these embryos by 5 days postfertilization, once morpholino degradation or dilution has occurred.

Dataset Information

A conserved role of ?A-crystallin in the development of the zebrafish embryonic lens.

Publications

A conserved role of αA-crystallin in the development of the zebrafish embryonic lens.

Similar Datasets

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