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Localization of mouse CLC-6 and CLC-7 mRNA and their functional complementation of yeast CLC gene mutant
1Graduate School, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan.
Abstract:
CLC-6 and CLC-7 belong to the family of voltage-dependent chloride channels. To learn more about the in vivo roles of CLC-6 and CLC-7, we performed in situ hybridization of these CLC channels in various mouse organs. Mouse CLC-6 (mCLC-6) was expressed in the peripheral region of seminiferous tubules in the testis, tracheal epithelium, epithelium of bronchioles, alveolar cells in the lung, acinar cells in the pancreas, and intestinal epithelium, but we could not detect signals from pancreatic islets. Mouse CLC-7 (mCLC-7) was expressed in neurons in the medulla oblongata, Purkinje cells in the cerebellum, proximal tubules in the kidney, and hepatocytes in the liver. The distribution of mCLC-6 and mCLC-7 were similar in the lung, pancreas, and testis. mCLC-6 functionally complemented the gef1 phenotype of a yeast strain in which a single CLC channel (GEF1) had been disrupted by homologous recombination. In contrast, mCLC-7 did not complement this gef1 phenotype. This study identified the cell types that express mCLC-6 and mCLC-7 in the mouse tissues, and the complementation assay suggested that mCLC-6 functions as an intracellular chloride channel.
Insights
This study maps the expression of chloride channel-6 (CLC-6) and chloride channel-7 (CLC-7) in mouse tissues. CLC-6 complements yeast gef1 mutations, suggesting it functions as an intracellular chloride channel.
Area of Science:
- Molecular Biology
- Cell Biology
- Physiology
Background:
- Voltage-dependent chloride channels, CLC-6 and CLC-7, play crucial roles in cellular functions.
- Understanding their in vivo distribution and function is essential for elucidating their physiological significance.
Purpose of the Study:
- To investigate the tissue-specific expression patterns of mouse CLC-6 (mCLC-6) and mouse CLC-7 (mCLC-7) in vivo.
- To functionally characterize mCLC-6 and mCLC-7 using a yeast complementation assay.
Main Methods:
- In situ hybridization was employed to determine the expression sites of mCLC-6 and mCLC-7 across various mouse organs.
- A yeast complementation assay was performed using a yeast strain with a disrupted GEF1 gene to assess the functional activity of mCLC-6 and mCLC-7.
Main Results:
- mCLC-6 expression was detected in the testis, lung, pancreas, and intestinal epithelium.
- mCLC-7 was found in the brain (medulla oblongata, cerebellum), kidney, and liver.
- mCLC-6, but not mCLC-7, functionally complemented the gef1 yeast phenotype, indicating a potential role as an intracellular chloride channel.
Conclusions:
- This study provides a comprehensive map of mCLC-6 and mCLC-7 expression in mouse tissues.
- The functional data suggest that mCLC-6 likely operates as an intracellular chloride channel, contributing to cellular chloride homeostasis.