Multiple transcripts of anoctamin genes expressed in the mouse submandibular salivary gland
한지혜, 김혜미, 서덕규, Lee Gene, 정의배, Yu Frank H.,
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한지혜 ( Han Ji-Hye ) - Seoul National University School of Dentistry Program in Neurobiology
김혜미 ( Kim Hye-Mi ) - Seoul National University School of Dentistry Program in Neurobiology
서덕규 ( Seo Deog-Gyu ) - Seoul National University School of Dentistry Department of Conservative Dentistry
( Lee Gene ) - Seoul National University School of Dentistry Department of Oral Biochemistry
정의배 ( Jeung Eui-Bae ) - Chungbuk National University College of Veterinary Medicine Laboratory of Veterinary Biochemistry
( Yu Frank H. ) - Seoul National University School of Dentistry Program in Neurobiology
KMID : 0363020150450020069
Abstract
Purpose: Salivary fluid formation is primarily driven by Ca2+-activated, apical efflux of chloride into the lumen of the salivary acinus. The anoctamin1 protein is an anion channel with properties resembling the endogenous calcium-activated chloride channels. In order to better understand the role of anoctamin proteins in salivary exocrine secretion, the expression of the ten members of the anoctamin gene family in the mouse submandibular gland was studied.
Methods: Total RNA extracted from mouse submandibular salivary glands was reverse transcribed using primer pairs to amplify the full-length coding regions of each anoctamin gene and was subcloned into plasmid vectors for DNA sequencing. Alternative splice variants were also screened by polymerase chain reaction using primer pairs that amplified six overlapping regions of the complementary DNA of each anoctamin gene, spanning multiple exons.
Results: Multiple anoctamin transcripts were found in the mouse submandibular salivary gland, including full-length transcripts of anoctamin1, anoctamin3, anoctamin4, anoctamin5, anoctamin6, anoctamin9, and anoctamin10. Exon-skipping splicing in the N-terminal exons of the anoctamins1, anoctamin5, and anoctamin6 genes resulted in multiple alternative splice variants. No expression of anoctamin2, anoctamin7, or anoctamin8 was found.
Conclusions: The predominant anoctamin transcript expressed in the mouse submandibular gland is anoctamin1ac. The chloride channel protein produced by anoctamin1ac is likely responsible for the Ca2+-activated chloride efflux, which is the rate-limiting step in salivary exocrine secretion.
키워드
Alternative splicing ; Chloride channels ; Submandibular gland
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