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PMID:18191123

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Citation

Yaung, J, Kannan, R, Wawrousek, EF, Spee, C, Sreekumar, PG and Hinton, DR (2008) Exacerbation of retinal degeneration in the absence of alpha crystallins in an in vivo model of chemically induced hypoxia. Exp. Eye Res. 86:355-65

Abstract

This study evaluated the role of crystallins in retinal degeneration induced by chemical hypoxia. Wild-type, alphaA-crystallin (-/-), and alphaB-crystallin (-/-) mice received intravitreal injection of 12 nmol (low dose), 33 nmol (intermediate dose) or 60 nmol (high dose) cobalt chloride (CoCl(2)). Hematoxylin and eosin and TdT-mediated dUTP nick-end labeling (TUNEL) stains were performed after 24 h, 96 h, and 1 week post-injection, while immunofluorescent stains for alphaA- and alphaB-crystallin were performed 1 week post-injection. The in vitro effects of CoCl(2) on alphaB-crystallin expression in ARPE-19 cells were determined by real time RT-PCR, Western blot, and confocal microscopy and studies evaluating subcellular distribution of alphaB-crystallin in the mitochondria and cytosol were also performed. Histologic studies revealed progressive retinal degeneration with CoCl(2) injection in wild-type mice. Retinas of CoCl(2) injected mice showed transient increased expression of HIF-1alpha which was maximal 24h after injection. Intermediate-dose CoCl(2) injection was associated with increased retinal immunofluorescence for both alphaA- and alphaB-crystallin; however, after high-dose injection, increased retinal degeneration was associated with decreased levels of crystallin expression. Injection of CoCl(2) at either intermediate or high dose in alphaA-crystallin (-/-) and alphaB-crystallin (-/-) mice resulted in much more severe retinal degeneration compared to wild-type eyes. A decrease in ARPE-19 total and cytosolic alphaB-crystallin expression with increasing CoCl(2) treatment and an increase in mitochondrial alphaB-crystallin were found. We conclude that lack of alpha-crystallins accentuates retinal degeneration in chemically induced hypoxia in vivo.

Links

PubMed PMC2731668 Online version:10.1016/j.exer.2007.11.007

Keywords

Animals; Apoptosis; Cells, Cultured; Disease Models, Animal; Disease Progression; Mice; Mice, Knockout; Mitochondria/metabolism; Pigment Epithelium of Eye/metabolism; Pigment Epithelium of Eye/pathology; Retinal Degeneration/chemically induced; Retinal Degeneration/metabolism; Retinal Degeneration/pathology; Reverse Transcriptase Polymerase Chain Reaction/methods; alpha-Crystallin B Chain/genetics; alpha-Crystallin B Chain/metabolism; alpha-Crystallins/deficiency; alpha-Crystallins/genetics; alpha-Crystallins/physiology

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

MOUSE:CRYAB

located_in

GO:0005829: cytosol

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:CRYAB

located_in

GO:0005739: mitochondrion

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:CRYAA

acts_upstream_of_or_within

GO:0001666: response to hypoxia

ECO:0000315: mutant phenotype evidence used in manual assertion

MGI:MGI:1858025

P

Seeded From UniProt

complete

MOUSE:CRYAA

acts_upstream_of_or_within

GO:0043066: negative regulation of apoptotic process

ECO:0000315: mutant phenotype evidence used in manual assertion

MGI:MGI:1858025

P

Seeded From UniProt

complete

MOUSE:CRYAB

acts_upstream_of_or_within

GO:0043066: negative regulation of apoptotic process

ECO:0000315: mutant phenotype evidence used in manual assertion

MGI:MGI:2652177

P

Seeded From UniProt

complete

MOUSE:CRYAB

part_of

GO:0005829: cytosol

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:CRYAB

part_of

GO:0005739: mitochondrion

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:CRYAB

acts_upstream_of_or_within

GO:0001666: response to hypoxia

ECO:0000315: mutant phenotype evidence used in manual assertion

MGI:MGI:2652177

P

Seeded From UniProt

complete


See also

References

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