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Alterations in Protein Expression in Tree Shrew Sclera during Development of Lens-induced Myopia and Recovery. Invest Ophthalmol Vis Sci. 2011 Oct 28; Authors: Frost MR, Norton TT Abstract Purpose
uring the development of, and recovery from, negative lens-induced myopia there is regulated remodeling of the scleral extracellular matrix (ECM) that controls the extensibility of the sclera. Difference Gel Electrophoresis (DIGE) was used to identify and categorize proteins whose levels are altered in this process.Methods:Two groups of 5 tree shrews started monocular lens wear 24 days after eye opening (days of visual experience [VE]). The lens-induced myopia (LIM) group wore a -5 D lens for 4 days. The recovery (REC) group wore a -5 D lens for 11 days and then recovered for 4 days. Two normal groups (28 and 39 days of VE; n=5 each) were also examined, age-matched to each of the treatment groups. Refractive and A-scan measures confirmed the effect of the treatments. Scleral proteins were isolated and resolved by DIGE. Proteins that differed in abundance were identified by mass spectrometry. Ingenuity Pathway Analysis was used to investigate potential biological pathway interactions.Results: During normal development (28 to 39 days of VE), eight proteins decreased and one protein increased in relative abundance. LIM-treated eyes were myopic and longer compared with control eyes; LIM-control eyes were slightly myopic compared with 28N eyes, indicating a yoking effect. In both the LIM-treated and LIM-control eyes there was a general down-regulation from normal of proteins involved in transcription, cell adhesion, and protein synthesis. Additional proteins involved in cell adhesion, actin cytoskeleton, transcriptional regulation, and ECM structural proteins, differed in the LIM-treated eyes vs. normal but did not differ in the control eyes vs. normal. REC-treated eyes were recovering from the induced myopia. The REC-control eye refractions were not significantly different from the 39N eyes and few proteins differed from age-matched normal eyes. The balance of protein expression in the REC-treated eyes, compared with normal eyes and REC-control eyes, shifted toward up-regulation or a return to normal levels of proteins involved in cell adhesion, cell division, cytoskeleton, and ECM structural proteins, including up-regulation of several cytoskeleton-related proteins not affected during myopia development.Conclusions:The DIGE procedure revealed new proteins whose abundance is altered during myopia development and recovery. Many of these are involved in cell-matrix adhesions, cytoskeleton and transcriptional regulation and extend our understanding of the remodeling that controls the extensibility of the sclera. Reductions in these proteins during minus lens wear may produce the increased scleral viscoelasticity that results in faster axial elongation. Recovery is not a mirror image of lens-induced myopia - many protein levels, decreased during LIM, returned to, or slightly above, normal and additional cytoskeleton proteins were up-regulated. However, no single protein or pathway appeared to be responsible for the scleral changes during myopia development or recovery. PMID: 22039233 [PubMed - as supplied by publisher]
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uring the development of, and recovery from, negative lens-induced myopia there is regulated remodeling of the scleral extracellular matrix (ECM) that controls the extensibility of the sclera. Difference Gel Electrophoresis (DIGE) was used to identify and categorize proteins whose levels are altered in this process.Methods:Two groups of 5 tree shrews started monocular lens wear 24 days after eye opening (days of visual experience [VE]). The lens-induced myopia (LIM) group wore a -5 D lens for 4 days. The recovery (REC) group wore a -5 D lens for 11 days and then recovered for 4 days. Two normal groups (28 and 39 days of VE; n=5 each) were also examined, age-matched to each of the treatment groups. Refractive and A-scan measures confirmed the effect of the treatments. Scleral proteins were isolated and resolved by DIGE. Proteins that differed in abundance were identified by mass spectrometry. Ingenuity Pathway Analysis was used to investigate potential biological pathway interactions.Results: During normal development (28 to 39 days of VE), eight proteins decreased and one protein increased in relative abundance. LIM-treated eyes were myopic and longer compared with control eyes; LIM-control eyes were slightly myopic compared with 28N eyes, indicating a yoking effect. In both the LIM-treated and LIM-control eyes there was a general down-regulation from normal of proteins involved in transcription, cell adhesion, and protein synthesis. Additional proteins involved in cell adhesion, actin cytoskeleton, transcriptional regulation, and ECM structural proteins, differed in the LIM-treated eyes vs. normal but did not differ in the control eyes vs. normal. REC-treated eyes were recovering from the induced myopia. The REC-control eye refractions were not significantly different from the 39N eyes and few proteins differed from age-matched normal eyes. The balance of protein expression in the REC-treated eyes, compared with normal eyes and REC-control eyes, shifted toward up-regulation or a return to normal levels of proteins involved in cell adhesion, cell division, cytoskeleton, and ECM structural proteins, including up-regulation of several cytoskeleton-related proteins not affected during myopia development.Conclusions:The DIGE procedure revealed new proteins whose abundance is altered during myopia development and recovery. Many of these are involved in cell-matrix adhesions, cytoskeleton and transcriptional regulation and extend our understanding of the remodeling that controls the extensibility of the sclera. Reductions in these proteins during minus lens wear may produce the increased scleral viscoelasticity that results in faster axial elongation. Recovery is not a mirror image of lens-induced myopia - many protein levels, decreased during LIM, returned to, or slightly above, normal and additional cytoskeleton proteins were up-regulated. However, no single protein or pathway appeared to be responsible for the scleral changes during myopia development or recovery. PMID: 22039233 [PubMed - as supplied by publisher] More...
