Abstract：
UNASSIGNED: To explore the functional consequences of two common variants, p.V37I and c.299-300delAT in hearing loss associated gene GJB2.
UNASSIGNED: Connexin 26 expression and gap junctional permeability were studied in HEK 293T cells transfected with plasmids expressing GJB2 wild-type, p.V37I, or c.299-300delAT CX26 proteins with fluorescent tags. Functional analyses of different GJB2 haplotypes were performed to fully assess the alteration of ionic and small-molecule coupling.
UNASSIGNED: The p.V37I protein was localized at the plasma membrane, but failed to effectively transport intercellular propidium iodide or Ca2+ efficiently, indicating impairment of both biochemical and ionic coupling. The presence of GJB2 p.V37I appeared to increase the sensitivity of cells to H2O2 treatment. In contrast, the known variant c.299-300delAT protein was not transported to the cell membrane and could not form gap junctions, instead being confined to the cytoplasm. Ionic and biochemical coupling was defective in c.299-300delAT-transfected cells.
UNASSIGNED: The p.V37I and c.299-300delAT GJB2 mutations resulted in deficient gap junction-mediated coupling. Environmental factors may impact the functional consequences of GJB2 p.V37I. These results may inspire the development of molecular therapies targeting GJB2 mutations for hearing loss.
UNASSIGNED: Connexin 26 expression and gap junctional permeability were studied in HEK 293T cells transfected with plasmids expressing GJB2 wild-type, p.V37I, or c.299-300delAT CX26 proteins with fluorescent tags. Functional analyses of different GJB2 haplotypes were performed to fully assess the alteration of ionic and small-molecule coupling.
UNASSIGNED: The p.V37I protein was localized at the plasma membrane, but failed to effectively transport intercellular propidium iodide or Ca2+ efficiently, indicating impairment of both biochemical and ionic coupling. The presence of GJB2 p.V37I appeared to increase the sensitivity of cells to H2O2 treatment. In contrast, the known variant c.299-300delAT protein was not transported to the cell membrane and could not form gap junctions, instead being confined to the cytoplasm. Ionic and biochemical coupling was defective in c.299-300delAT-transfected cells.
UNASSIGNED: The p.V37I and c.299-300delAT GJB2 mutations resulted in deficient gap junction-mediated coupling. Environmental factors may impact the functional consequences of GJB2 p.V37I. These results may inspire the development of molecular therapies targeting GJB2 mutations for hearing loss.
摘要：
■为了探索两种常见变体的功能后果，p.V37I和c.299-300delAT在听力损失相关基因GJB2中的表达。
■在转染表达GJB2野生型质粒的HEK293T细胞中研究了连接蛋白26的表达和间隙连接通透性，p.V37I,或c.299-300delATCX26蛋白与荧光标签。进行了不同GJB2单倍型的功能分析，以全面评估离子和小分子偶联的改变。
■p.V37I蛋白定位于质膜，但未能有效转运细胞间碘化丙啶或Ca2+，表明生化和离子耦合的损害。GJB2p.V37I的存在似乎增加了细胞对H2O2处理的敏感性。相比之下,已知的变异c.299-300delAT蛋白不被转运到细胞膜上，不能形成间隙连接，而是局限于细胞质。在c.299-300delAT转染的细胞中，离子和生化偶联存在缺陷。
■p.V37I和c.299-300delATGJB2突变导致间隙连接介导的偶联缺陷。环境因素可能会影响GJB2p.V37I.的功能后果。这些结果可能会激发针对听力损失的GJB2突变的分子疗法的发展。
■在转染表达GJB2野生型质粒的HEK293T细胞中研究了连接蛋白26的表达和间隙连接通透性，p.V37I,或c.299-300delATCX26蛋白与荧光标签。进行了不同GJB2单倍型的功能分析，以全面评估离子和小分子偶联的改变。
■p.V37I蛋白定位于质膜，但未能有效转运细胞间碘化丙啶或Ca2+，表明生化和离子耦合的损害。GJB2p.V37I的存在似乎增加了细胞对H2O2处理的敏感性。相比之下,已知的变异c.299-300delAT蛋白不被转运到细胞膜上，不能形成间隙连接，而是局限于细胞质。在c.299-300delAT转染的细胞中，离子和生化偶联存在缺陷。
■p.V37I和c.299-300delATGJB2突变导致间隙连接介导的偶联缺陷。环境因素可能会影响GJB2p.V37I.的功能后果。这些结果可能会激发针对听力损失的GJB2突变的分子疗法的发展。
