Abstract
BackgroundMouse models have served a valuable role in deciphering various facets of Salivary Gland (SG) biology, from normal developmental programs to diseased states. To facilitate such studies, gene expression profiling maps have been generated for various stages of SG organogenesis. However these prior studies fall short of capturing the transcriptional complexity due to the limited scope of gene-centric microarray-based technology. Compared to microarray, RNA-sequencing (RNA-seq) offers unbiased detection of novel transcripts, broader dynamic range and high specificity and sensitivity for detection of genes, transcripts, and differential gene expression. Although RNA-seq data, particularly under the auspices of the ENCODE project, have covered a large number of biological specimens, studies on the SG have been lacking.ResultsTo better appreciate the wide spectrum of gene expression profiles, we isolated RNA from mouse submandibular salivary glands at different embryonic and adult stages. In parallel, we processed RNA-seq data for 24 organs and tissues obtained from the mouse ENCODE consortium and calculated the average gene expression values. To identify molecular players and pathways likely to be relevant for SG biology, we performed functional gene enrichment analysis, network construction and hierarchal clustering of the RNA-seq datasets obtained from different stages of SG development and maturation, and other mouse organs and tissues. Our bioinformatics-based data analysis not only reaffirmed known modulators of SG morphogenesis but revealed novel transcription factors and signaling pathways unique to mouse SG biology and function. Finally we demonstrated that the unique SG gene signature obtained from our mouse studies is also well conserved and can demarcate features of the human SG transcriptome that is different from other tissues.ConclusionsOur RNA-seq based Atlas has revealed a high-resolution cartographic view of the dynamic transcriptomic landscape of the mouse SG at various stages. These RNA-seq datasets will complement pre-existing microarray based datasets, including the Salivary Gland Molecular Anatomy Project by offering a broader systems-biology based perspective rather than the classical gene-centric view. Ultimately such resources will be valuable in providing a useful toolkit to better understand how the diverse cell population of the SG are organized and controlled during development and differentiation.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3228-7) contains supplementary material, which is available to authorized users.
Highlights
Mouse models have served a valuable role in deciphering various facets of Salivary Gland (SG) biology, from normal developmental programs to diseased states
Transcriptomic map of the mouse salivary gland during development and adult In order to better define the dynamic changes in global gene expression levels and to identify new tissue-specific and tissue-enriched regulators, we performed RNA-seq based expression profiling of the mouse submandibular salivary gland at various key stages of embryonic development in addition to post-natal and adult tissues
We isolated total RNA from the mouse submandibular gland dissected from E14.5 day old embryos (Pseudoglandular stage - representing the onset of branching morphogenesis), E16.5 (Canalicular stage representing the onset of cytodifferentiation) and E18.5 (Terminal Bud stage - representing the expansion of acini and lumenization of ducts and acini) [1, 4]
Summary
Mouse models have served a valuable role in deciphering various facets of Salivary Gland (SG) biology, from normal developmental programs to diseased states To facilitate such studies, gene expression profiling maps have been generated for various stages of SG organogenesis. As the embryo develops to E14, the gland undergoes rapid proliferation and intricate branching morphogenesis, during which the end buds undergo successive rounds of clefting resulting in the generation of multiple epithelial buds. This Pseudoglandular stage coincides with reorganization of the end buds and the formation of the acini, which are the main secretory units of the salivary gland. While the gland is functional and able to secrete saliva at birth, further acinar maturation and differentiation continues postnatally, and by puberty, differentiation of the granular convoluted tubules is complete [1, 7]
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