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Integrative analysis of transcriptome and proteome sheds light on functional differences between human salivary glands

ABSTRACT: Saliva aids in the predigestion of food and perception of taste. It helps to maintain the integrity of the mineralized tooth and epithelial surfaces in the mouth, and shields the oro-digestive tract from environmental hazards and invading pathogens. Although salivary glands and saliva fluid are biologically and functionally inseparable, they have thus far been investigated as separate entities. To bridge this gap, we performed an integrative analysis of the transcriptome of 27 samples collected from the major human adult and fetal major salivary glands - submandibular, sublingual, and parotid - along with mass-spectrometry-based saliva proteome data and immunohistochemical localization in glandular tissue. Our results suggest that functional maturation at the transcriptome level occurs late in gland development, and is driven mainly by the transcription of genes that code for secreted saliva proteins. We further provide evidence that protein dosage of the most abundant salivary proteins secreted by the salivary glands is predominantly regulated at the transcriptome level. Finally, we demonstrate distinct transcriptomic profiles of each major salivary gland type that reveal functional specialization and will aid in future clinical analyses. Our study provides the hitherto most comprehensive RNAseq dataset of healthy salivary glands in humans, thus establishing a robust framework for deeper studies of saliva and salivary gland biology, development, and evolution, ultimately paving the way for better understanding the importance of these craniofacial secretory organs in health and their malfunctions in disease.

ORGANISM(S): Homo sapiens

PROVIDER: GSE143702 | GEO | 2020/07/10

REPOSITORIES: GEO

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Project description:MyD88 is an important adaptor protein for signal transduction downstream of Toll-like receptors and TACI, receptors for regulation of innate immunity and B cell responses, respectively. The surfaces of oral mucosa are protected from infections by antimicrobial proteins and natural immunoglobulins that are constantly secreted in saliva, serving as principal innate immune defense in the oral cavity. Although MyD88-mediated signaling has a regulatory role in the intestinal mucosal immunity, its specific role in the oral cavity has been remained elusive. To identify the specific roles of MyD88-dependent signaling in gene expression profiles in salivary glands, we performed microarray analysis of the major salivary gland tissues (submandibular gland plus sublingual gland) from control (wild-type C57BL/6) mice and C57BL/6 background Myd88-null mice using Agilent Whole Mouse Genome Oligo Microarrays (8x60K, Design ID 028005). Total RNA was extracted from the major salivary gland tissues (submandibular gland plus sublingual gland) of wild-type and Myd88-null mice using using Trizol reagent (Life Technologies, Rockville, MD, USA) according to the manufacturer's instructions. 100 ng of all RNA samples was used as template to produce Cy3-labeled cRNA. The RNA samples were amplified and labeled using the Low Input Quick Amp Labeling Kit (Agilent Technologies, Palo Alto, USA) following the manufacturerM-bM-^@M-^Ys protocol. Cy3 labeled cRNAs were hybridized overnight (17 hours, 65M-BM-0C) to an Agilent Whole Mouse Genome Oligo Microarrays (8x60K, Design ID 028005) using AgilentM-bM-^@M-^Ys recommended hybridization chamber and oven.Finally, microarrays were washed once with wash buffer 1 for 1 min at RT followed by a second wash with wash buffer 2 for 1 min at 37M-BM-0C and a final wash step with acetonitrile for 30 sec at RT. Fluorescence signals of the hybridized Agilent Microarrays were detected using AgilentM-bM-^@M-^Ys Microarray Scanner System (G2505C, Agilent Technologies). The Agilent Feature Extraction Software (FES 10.7.3.1 ) was used to read out and process the microarray image files.

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Integrative analysis of transcriptome and proteome sheds light on functional differences between human salivary glands

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