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RNA-seq of the E14.5 lacrimal gland

ABSTRACT: The goal of this study is to find the transcriptional targets downstream of PI3K signaling during lacrimal gland development. We performed laser capture microdissection of the lacrimal gland epithelial tissue at embryonic day E14.5 from control embryos and mutant embryos containing lacrimal gland-specific deletion of PI3K subunits. After tissue harvest, RNA was extracted, conversion to cDNA and amplification was performed by Clontech SMART-seq v4 Ultra low input RNA kit,and cDNA library construction was performed using Nextera XT DNA library preparation kit by core facility at Columbia university prior to RNA sequencing. After preparation of cDNA library, each sample was sequenced using Illumina platform

ORGANISM(S): Mus musculus

PROVIDER: GSE158300 | GEO | 2020/09/22

REPOSITORIES: GEO

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RNA-seq of the lacrimal gland epithelial tissue in control vs Pea3-depleted tissue in mouse embyro

Project description:The goal of the study is to find the transcriptional targets downstream of Pea3-mediated signaling in the lacrimal gland epithelium. In order to look for the potential targets, we performed laser capture microdissection of the lacrimal gland epithelial tissue at embryonic day E14.5 from control embryos as well as mutant embryos containing epithelium-specific deletion of Pea3. After tissue harvest, RNA was extracted. Conversion to cDNA as well as amplification was performed by Clontech SMART-seq v4 Ultra low input RNA kit, as well as cDNA library construction was performed using Nextera XT DNA library preparation kit by core facility at Columbia university prior to RNA sequencing. After preparation of cDNA library, each sample was sequenced using Illumina platform.

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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.

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miR-205 is a critical regulator of lacrimal gland development

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