Impact of α-crystallin protein loss on zebrafish lens development.
CRISPR
Cataract
Crystallins
Lens
Zebrafish
Journal
Experimental eye research
ISSN: 1096-0007
Titre abrégé: Exp Eye Res
Pays: England
ID NLM: 0370707
Informations de publication
Date de publication:
02 2023
02 2023
Historique:
received:
03
10
2022
revised:
27
11
2022
accepted:
19
12
2022
pmc-release:
01
02
2024
pubmed:
27
12
2022
medline:
15
2
2023
entrez:
26
12
2022
Statut:
ppublish
Résumé
The α-crystallin small heat shock proteins contribute to the transparency and refractive properties of the vertebrate eye lens and prevent the protein aggregation that would otherwise produce lens cataracts, the leading cause of human blindness. There are conflicting data in the literature as to what role the α-crystallins may play in early lens development. In this study, we used CRISPR gene editing to produce zebrafish lines with mutations in each of the three α-crystallin genes (cryaa, cryaba and cryabb) to prevent protein production. The absence of each α-crystallin protein was analyzed by mass spectrometry, and lens phenotypes were assessed with differential interference contrast microscopy and histology. Loss of αA-crystallin produced a variety of lens defects with varying severity in larvae at 3 and 4 dpf but little substantial change in normal fiber cell denucleation. Loss of αBa-crystallin produced no substantial lens defects. Our cryabb mutant produced a truncated αBb-crystallin protein and showed no substantial change in lens development. Mutation of each α-crystallin gene did not alter the mRNA levels of the remaining two, suggesting a lack of genetic compensation. These data suggest that αA-crystallin plays some role in lens development, but the range of phenotype severity in null mutants indicates its loss simply increases the chance for defects and that the protein is not essential. Our finding that cryaba and cryabb mutants lack noticeable lens defects is congruent with insubstantial transcript levels for these genes in lens epithelial and fiber cells through five days of development. Future experiments can explore the molecular mechanisms leading to lens defects in cryaa null mutants and the impact of αA-crystallin loss during zebrafish lens aging.
Identifiants
pubmed: 36572168
pii: S0014-4835(22)00439-0
doi: 10.1016/j.exer.2022.109358
pmc: PMC9918708
mid: NIHMS1861648
pii:
doi:
Substances chimiques
alpha-Crystallins
0
Crystallins
0
alpha-Crystallin A Chain
0
Proteins
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
109358Subventions
Organisme : NEI NIH HHS
ID : P30 EY010572
Pays : United States
Organisme : NEI NIH HHS
ID : R15 EY013535
Pays : United States
Informations de copyright
Copyright © 2022 Elsevier Ltd. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of competing interest None.
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