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A time course microarray study of gene expression in the mouse lacrimal gland after acute corneal trauma.

ABSTRACT: Examine the effect of stimulation stress produced by chemical burn to the mice cornea in the lacrimal gland over a time course (0.5 - 360 hours) in comparison with the non-stimulated control animals. Using silver nitrate applicator (silver nitrate 37.5 mg and potassium nitrate 12.5 mg, Graham-Field,Inc.,) do two burns per eye, each with a separate applicator. For animals observed for more than 24 hours, eyes were then treated with gentamicin ointment to minimize infection. Total RNA were extracted from both burn and control eyes and were covalently coupled separately with Cy5 and Cy3 monoreactive fluors. The Cy5 and Cy3 labelled RNA were hybridized to the microarray. Fluorescent array images were collected for both Cy3 and Cy5 with ScanArray 4000XL and image intensity data were extracted and analyzed with Imagene analysis software. Ref:Fang Y, Choi D, Searles RP, Mathers WD.A time course microarray study of gene expression in the mouse lacrimal gland after acute corneal trauma.Invest Ophthalmol Vis Sci. 2005 Feb;46(2):461-9. Keywords = Corneal injury Keywords = mouse Keywords = lacrimal gland Keywords = gene microarrays Keywords: time-course

ORGANISM(S): Mus musculus

PROVIDER: GSE1393 | GEO | 2004/05/21

SECONDARY ACCESSION(S): PRJNA90175

REPOSITORIES: GEO

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Project description:Purpose: Identify the differentially expressed circular RNA (circRNA) and elucidate their potential function in AQP5 knockout (AQP5-/-) mice with primary dry eye phenotype. Methods: A slit lamp examination was performed on AQP5 knockout mice to assess corneal epithelial defect by fluorescein sodium staining. Hematoxylin-eosin staining and transmission electron microscope analysis were performed to access the structure of lacrimal gland epithelial cells. The expression profiles of circRNA and messenger RNA (mRNA) were determined by microarray analysis. The selected circRNA was verified by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were used to predict biological functions and potential pathways of parental genes involved in lacrimal gland epithelial cell changes. According to the bioinformatics analysis of identified circRNA, we can predict a circRNA-miRNA-mRNA network. Results: AQP5-/- mice exhibit spontaneous dry eye symptoms. AQP5 deficiency obviously changes the structure of lacrimal gland epithelial cells. Analysis showed that compared to AQP5+/+ mice, 30 circRNAs in the lacrimal glands of AQP5‑/- mice had differential expression (fold change ≥ 2.0, P < 0.05). Nine upregulated circRNAs were identified by qRT-PCR; nine upregulated validated circRNAs, 40 altered microRNAs (miRNAs), and nine upregulated mRNAs were involved in the network analysis. KEGG analysis showed these nine target genes were expressed in phagosomes. Conclusions: AQP5-/- mice have primary and stable dry eye phenotypes from birth. The study identified different expressed circRNAs in lacrimal glands between AQP5-/- and AQP5+/+ mice, predicting a circRNA-miRNA-mRNA network of phagosomes. CircRNA likely plays an important role in lacrimal gland epithelial cell pathogenesis. Therefore, it is reasonable to use circRNA as a potential therapeutic target for dry eyes.

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A time course microarray study of gene expression in the mouse lacrimal gland after acute corneal trauma.

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