Dataset Information

Retinal proteome alterations in a mouse model of type 2 diabetes.

ABSTRACT:

Aims/hypothesis

Diabetic retinopathy is a major complication of type 2 diabetes and the leading cause of blindness in adults of working age. Neuronal defects are known to occur early in disease, but the source of this dysfunction is unknown. The aim of this study was to examine differences in the retinal membrane proteome among non-diabetic mice and mouse models of diabetes either with or without metformin treatment.

Methods

Alterations in the retinal membrane proteome of 10-week-old diabetic db/db mice, diabetic db/db mice orally treated with the anti-hyperglycaemic metformin, and congenic wild-type littermates were examined using label-free mass spectrometry. Pathway enrichment analysis was completed with Genomatix and Ingenuity. Alterations in Slc17a7 mRNA and vesicular glutamate transporter 1 (VGLUT1) protein expression were evaluated using real-time quantitative PCR and IMMUNOFLUORESCENCE.

Results

A total of 98 proteins were significantly differentially abundant between db/db and wild-type animals. Pathway enrichment analysis indicated decreases in levels of proteins related to synaptic transmission and cell signalling. Metformin treatment produced 63 differentially abundant proteins compared with untreated db/db mice, of which only 43 proteins were found to occur in both datasets, suggesting that treatment only partially normalises the alterations induced by diabetes. VGLUT1, which is responsible for loading glutamate into synaptic vesicles, was found to be differentially abundant in db/db mice and was not normalised by metformin. The decrease in Slc17a7/VGLUT1 was confirmed by transcriptomic and immunocytochemical analysis.

Conclusions/interpretation

These findings expand the knowledge of the protein changes in diabetic retinopathy and suggest that membrane-associated signalling proteins are susceptible to changes that are partially ameliorated by treatment

SUBMITTER: Ly A

PROVIDER: S-EPMC3855476 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature

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Publications

Retinal proteome alterations in a mouse model of type 2 diabetes.

Ly Alice A Scheerer Markus F MF Zukunft Sven S Muschet Caroline C Merl Juliane J Adamski Jerzy J de Angelis Martin Hrabě MH Neschen Susanne S Hauck Stefanie M SM Ueffing Marius M

Diabetologia 20140101 1

<h4>Aims/hypothesis</h4>Diabetic retinopathy is a major complication of type 2 diabetes and the leading cause of blindness in adults of working age. Neuronal defects are known to occur early in disease, but the source of this dysfunction is unknown. The aim of this study was to examine differences in the retinal membrane proteome among non-diabetic mice and mouse models of diabetes either with or without metformin treatment.<h4>Methods</h4>Alterations in the retinal membrane proteome of 10-week- ...[more]

PMID: 24078137

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Project description:Aims/hypothesisDiabetic retinopathy is increasing in prevalence worldwide and is fast becoming a global epidemic and a leading cause of visual loss. Current therapies are limited, and the development of effective treatments for diabetic retinopathy requires a greater in-depth knowledge of disease progression and suitable modelling of diabetic retinopathy in animals. The aim of this study was to assess the early pathological changes in retinal morphology and neuronal, inflammatory and vascular features consistent with diabetic retinopathy in the ob/ob mouse model of type 2 diabetes, to investigate whether features similar to those in human diabetic retinopathy were present.MethodsMale and female wild-type (+/+), heterozygous (+/-) and homozygous (-/-) BTBR ob/ob mice were examined at 6, 10, 15 and 20 weeks of age. Animals were weighed and blood glucose was measured. TUNEL and brain-specific homeobox/POU domain protein 3A (BRN3A) markers were used to examine retinal ganglion cells. We used immunostaining (collagen IV and platelet endothelial cell adhesion molecule [PECAM]/CD31) to reveal retinal vessel degeneration. Spectral domain optical coherence tomography was used to reveal changes in the thickness and structure of the retinal layer. Vitreous fluorophotometry was used to investigate vascular permeability. A-waves, b-waves and oscillatory potentials were measured under photopic and scotopic conditions. Concanavalin A leucostasis and immunostaining with glial fibrillary acidic protein (GFAP) and ionised calcium-binding adapter molecule 1 (IBA-1) identified differences in inflammatory status. Paraffin sections and transmission electron microscopy were used to reveal changes in the thickness and structure of the retinal layer.ResultsFollowing the development of obesity and hyperglycaemia in 2-week-old and 3-week-old ob-/ob- mice, respectively (p?<?0.001), early functional deficits (p?<?0.001) and thinning of the inner retina (p?<?0.001) were identified. Glial activation, leucostasis (p?<?0.05) and a shift in microglia/macrophage phenotype were observed before microvascular degeneration (p?<?0.05) and elevated vascular permeability occurred (p?<?0.05).Conclusions/interpretationThe present characterisation of the development of diabetic retinopathy in the ob/ob mouse represents a platform that will enable the development of new therapies, particularly for the early stages of disease.

Dataset Information

Retinal proteome alterations in a mouse model of type 2 diabetes.

Aims/hypothesis

Methods

Results

Conclusions/interpretation

Publications

Retinal proteome alterations in a mouse model of type 2 diabetes.

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