Premium
Global microarray analysis and metabolic pathway profiling of a transgenic model of conditional, selective Müller cell ablation
Author(s) -
CHUNG SH,
SHEN W,
YANG J,
JAYAWARDANA K,
GILLIES MC
Publication year - 2013
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2013.2765.x
Subject(s) - biology , microarray analysis techniques , downregulation and upregulation , microbiology and biotechnology , laser capture microdissection , gene expression profiling , microarray , cell , gene , gene expression , genetics
Purpose Müller cells, the principal glia cells in mammalian retina, play a critical role in retinal homeostasis. We have developed a transgenic Cre‐Lox model for selective, conditional Müller cell ablation to examine the relationship between Müller cell dysfunction and retinal diseases. The purpose of this study was to examine differentially expressed genes and their related pathways with microarray analysis, then to profile metabolic pathways in areas of Müller cell ablation. Methods Affymatrix microarray was performed on whole retina samples 1 week, 1 month and 3 months after induced Müller cell ablation. Data were analysed with limma package (p<0.05) and qRT‐PCR was used for array validation. Isolation of patches of Müller cell ablation was achieved by laser capture microdissection (LCM) and qRT‐PCR was conducted on pathway related genes. Immunofluorescence microscopy was used to validate results. Results Neuroprotection and apoptosis‐related genes were upregulated 1 week after Müller cell ablation, angiogenesis, tight junction and metabolic‐pathway related genes were downregulated later. Further analysis of glycolytic and mTOR pathways with tissue obtained by LCM revealed significant downregulation of genes related to these pathways in patches of Müller cell loss compared with controls. Immunofluorescent studies revealed that the dowregulations mainly occurred in the photoreceptor segments, although Enolase1 colocalised with Müller cells. Conclusion We found reduction of transcription and expression of proteins involved in key metabolic pathways in patches of Müller cell ablation. This study provides new insights into the relationship between Müller cell dysfunction and retinal diseases.