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The submandibular and sublingual glands maintain oral microbial homeostasis through multiple antimicrobial proteins

ABSTRACT: Oral microbial homeostasis is a key factor affecting oral health, and saliva plays a significant role in maintaining oral microbial homeostasis. The submandibular gland (SMG) and sublingual gland (SLG) together produce the most saliva at rest. Organic ingredients, including antimicrobial proteins, are rich and distinctive and depend on the type of acinar cells in the SMG and SLG. However, the functions of the SMG and SLG in maintaining oral microbial homeostasis have been difficult to identify and distinguish, given their unique anatomical structures. Therefore, we analyzed each gland using single-cell RNA sequencing.

ORGANISM(S): Mus musculus

PROVIDER: GSE216476 | GEO | 2023/01/03

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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The submandibular and sublingual glands maintain oral microbial homeostasis through multiple antimicrobial proteins

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