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Research output: Contribution to Journal/Magazine › Journal article › peer-review
A CNS-specific hypomorphic Pdgfr-beta mutant model of diabetic retinopathy. / Jadeja, Shalini; Mort, Richard L.; Keighren, Margaret et al.
In: Investigative Ophthalmology and Visual Science, Vol. 54, No. 5, 01.05.2013, p. 3569-3578.Research output: Contribution to Journal/Magazine › Journal article › peer-review
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TY - JOUR
T1 - A CNS-specific hypomorphic Pdgfr-beta mutant model of diabetic retinopathy
AU - Jadeja, Shalini
AU - Mort, Richard L.
AU - Keighren, Margaret
AU - Hart, Alan W.
AU - Joynson, Russell
AU - Wells, Sara
AU - Potter, Paul K.
AU - Jackson, Ian J.
PY - 2013/5/1
Y1 - 2013/5/1
N2 - PURPOSE: A mouse mutant identified during a recessive N-ethyl-N-nitrosourea (ENU) mutagenesis screen exhibited ocular hemorrhaging resulting in a blood-filled orbit, and hence was named "redeye." We aimed to identify the causal mutation in redeye, and evaluate it as a model for diabetic retinopathy (DR).METHODS: The causative gene mutation in redeye was identified by haplotype mapping followed by exome sequencing. Glucose tolerance tests, detailed histologic and immunofluorescence analyses, and vascular permeability assays were performed to determine the affect of redeye on glucose metabolism, pericyte recruitment, and the development of the retinal vasculature and blood-retinal barrier (BRB).RESULTS: A mutation was identified in the Pdgfrb gene at position +2 of intron 6. We show that this change causes partial loss of normal splicing resulting in a frameshift and premature termination, and, therefore, a substantial reduction in normal Pdgfrb transcript. The animals exhibit defective pericyte recruitment restricted to the central nervous system (CNS) causing basement membrane and vascular patterning defects, impaired vascular permeability, and aberrant BRB development, resulting in vascular leakage and retinal ganglion cell apoptosis. Despite exhibiting classic features of diabetic retinopathy, redeye glucose tolerance is normal.CONCLUSIONS: The Pdgfrb(redeye/redeye) mice exhibit all of the features of nonproliferative DR, including retinal neurodegeneration. In addition, the perinatal onset of the CNS-specific vascular phenotype negates the need to age animals or manage diabetic complications in other organs. Therefore, they are a more useful model for diseases involving pericyte deficiencies, such as DR, than those currently being used.
AB - PURPOSE: A mouse mutant identified during a recessive N-ethyl-N-nitrosourea (ENU) mutagenesis screen exhibited ocular hemorrhaging resulting in a blood-filled orbit, and hence was named "redeye." We aimed to identify the causal mutation in redeye, and evaluate it as a model for diabetic retinopathy (DR).METHODS: The causative gene mutation in redeye was identified by haplotype mapping followed by exome sequencing. Glucose tolerance tests, detailed histologic and immunofluorescence analyses, and vascular permeability assays were performed to determine the affect of redeye on glucose metabolism, pericyte recruitment, and the development of the retinal vasculature and blood-retinal barrier (BRB).RESULTS: A mutation was identified in the Pdgfrb gene at position +2 of intron 6. We show that this change causes partial loss of normal splicing resulting in a frameshift and premature termination, and, therefore, a substantial reduction in normal Pdgfrb transcript. The animals exhibit defective pericyte recruitment restricted to the central nervous system (CNS) causing basement membrane and vascular patterning defects, impaired vascular permeability, and aberrant BRB development, resulting in vascular leakage and retinal ganglion cell apoptosis. Despite exhibiting classic features of diabetic retinopathy, redeye glucose tolerance is normal.CONCLUSIONS: The Pdgfrb(redeye/redeye) mice exhibit all of the features of nonproliferative DR, including retinal neurodegeneration. In addition, the perinatal onset of the CNS-specific vascular phenotype negates the need to age animals or manage diabetic complications in other organs. Therefore, they are a more useful model for diseases involving pericyte deficiencies, such as DR, than those currently being used.
KW - Animals
KW - Base Sequence
KW - Basement Membrane
KW - Blood-Retinal Barrier
KW - Codon, Nonsense
KW - Diabetic Retinopathy
KW - Disease Models, Animal
KW - Exons
KW - Female
KW - Frameshift Mutation
KW - Glucose Tolerance Test
KW - Haplotypes
KW - Introns
KW - Male
KW - Mice
KW - Mice, Inbred C3H
KW - Mice, Mutant Strains
KW - Molecular Sequence Data
KW - Mutagenesis
KW - Pericytes
KW - RNA Splice Sites
KW - Receptor, Platelet-Derived Growth Factor beta
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1167/iovs.12-11125
DO - 10.1167/iovs.12-11125
M3 - Journal article
C2 - 23633653
VL - 54
SP - 3569
EP - 3578
JO - Investigative Ophthalmology and Visual Science
JF - Investigative Ophthalmology and Visual Science
SN - 0146-0404
IS - 5
ER -