Project description: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
Project description:We identified that Sparc gene expression is upregulated in corneal epithelial cells in a mouse model of dry eye disease involving lacrimal gland excision. Therefore, in this experiment we assess the effect of SPARC treatment on the transcriptome of human corneal epithelial cells.
Project description:Background: The study of human lacrimal gland biology and development is limited. Lacrimal gland tissue is damaged or poorly functional in a number of disease states including dry eye disease. Development of cell based therapies for lacrimal gland diseases requires a better understanding of the gene expression and signaling pathways in lacrimal gland. Differential gene expression analysis between lacrimal gland and other embryologically similar tissues may be helpful in furthering our understanding of lacrimal gland development. Methods: We performed global gene expression analysis of human lacrimal gland tissue using Affymetrix ® gene expression arrays. Primary data from our laboratory was compared with datasets available in the NLM GEO database for other surface ectodermal tissues including salivary gland, skin, conjunctiva and corneal epithelium. Results: The analysis revealed statistically significant difference in the gene expression of lacrimal gland tissue compared to other ectodermal tissues. The lacrimal gland specific, cell surface secretory protein encoding genes and critical signaling pathways which distinguish lacrimal gland from other ectodermal tissues are described. Conclusions: Differential gene expression in human lacrimal gland compared with other ectodermal tissue types revealed interesting patterns which may serve as the basis for future studies in directed differentiation among other areas.
Project description:PACAP is a neuropeptide that promotes lacrimal fluid secretion, but its direct action on corneal injury remains to be clarified. Therefore, the effect of PACAP on corneal epithelial repair was clarified using mouse corneal wound-injury model and human corneal epithelial culture cells. PAC1-R mRNA and its immunoreactivity was detected in mouse and corneal epithelium. In corneal wound-injury model mice, PACAP eye drop significantly reduced the injured area at 12 hours, and the effect was cancelled by co-treatment with the PACAP receptor antagonist. PACAP heterozygous and PAC1-R knockout mouse delayed the corneal healing. Although surgical removal of the lacrimal gland attenuates corneal healing, PACAP eye drop on the eyes significantly recuperated corneal damage. In an in-vitro study, PACAP treatment in human corneal epithelial cells significantly decreased the injury area induced by scratching. PACAP treatment in the in-vitro human corneal epithelial cell experiments significantly reduced the area of injury and abolished the corneal repair effect due to the inhibitory effect of Ara-C on proliferation. DNA whole-genome microarray analysis suggested that the nuclear receptor NR4A1 is an important factor in corneal epithelial proliferation, and THPN, which induces nuclear export of NR4A1, suppressed PACAP-induced proliferation of human corneal epithelial cells and the repair effect of mouse corneal epithelium. These data suggest that PACAP stimulates corneal repair by corneal epithelial proliferation via PAC1-R and NR4A1 transcriptional activity. PACAP could be a good candidate as an eye-drop medication for corneal injury disease including dry eye syndrome.
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.
Project description:To analyze the gene expression of mouse embryonic lacrimal gland and its developmental related organs such as harderian gland and eye lid. We performed microarray using them.
Project description:The Effect of Sex on Lacrimal Gland Gene Expression in the MRL/lpr and Non-obese Diabetic Mouse Models of SjšgrenÕs Syndrome Keywords: Female vs Male Lacrimal Gland Gene Expression. Female and male lacrimal glands were harvested each mouse strain . Tissues were pooled into 3 biological replicates and were hybridized to separate microarrays. Each cRNA prep was hybridized to a GE Healthcare/Amersham Biosciences CodeLink UniSet Mouse 20K I Bioarray and a Affymetrix GeneChip Mouse Expression Array 430A.
Project description:Advanced age is one of the most recognizable risk factors for dry eye. Dry eye disease affects millions worldwide and can result from age-related lacrimal gland dysfunction, which correlates with a decline in lacrimal gland secretory cell function and chronic inflammation. This study investigated the potential of calorie restriction to maintain LG and ocular surface health. Adult female C57BL/6J mice were subjected to a 40% calorie restriction for four months, starting at 6–7 months and continuing until 10–11 months. These mice were compared to controls fed ad libitum. Bulk RNA sequencing of lacrimal glands, conjunctiva and cornea subjected to calorie restriction compared to ad libitum revealed significant differentially expressed genes (DEGs). Pathways enriched in the upregulated DEGs indicate enhanced circadian rhythm, secretory functions and lipid metabolism. These findings were confirmed using individual qPCR reactions and western blotting. In contrast, pathways enriched in the downregulated DEGs were associated with immune cell activation, adaptive immune responses, extracellular matrix remodeling, and metalloproteinase activity. Histological sections of calorie-restricted lacrimal glands revealed reduced mononuclear cell infiltration and fewer positive cells for CD4, CD19, and MHC II compared to libitum lacrimal glands. Calorie restriction also prevented age-related corneal barrier dysfunction and mitigated age-related conjunctival goblet cell loss, hallmarks of dry eye disease. These findings suggest that calorie restriction supports lacrimal gland and ocular surface health by reducing inflammation and extracellular matrix remodeling and by enhancing the lacrimal glands secretory function.
Project description:The Effect of Sex on Lacrimal Gland Gene Expression in the MRL/lpr and Non-obese Diabetic Mouse Models of SjšgrenÕs Syndrome Keywords: Female vs Male Lacrimal Gland Gene Expression.
Project description:First, we determined the gene expression profile of WT mouse lacrimal gland tissue and organoids in expansion and in differentiation. Then, we sought the differences between WT and Pax6-KO mouse lacrimal gland organoids in expansion and in differentiation.