Cells utilize calcium ions(Ca^2+)to signal almost all aspects of cellular life,ranging from cell proliferation to cell death,in a spatially and temporally regulated manner.A key aspect of this regulation is the compar...Cells utilize calcium ions(Ca^2+)to signal almost all aspects of cellular life,ranging from cell proliferation to cell death,in a spatially and temporally regulated manner.A key aspect of this regulation is the compartmen-talization of Ca^2+in various cytoplasmic organelles that act as intracellular Ca^2+stores.Whereas Ca^2+release from the large-volume Ca^2+stores,such as the endoplasmic reticulum(ER)and Golgi apparatus,are preferred for signal transduction,Ca^2+release from the small-volume individual vesicular stores that are dispersed throughout the cell,such as lysosomes,may be more useful in local regulation,such as membrane fusion and individualized vesicular movements.Conceivably,these two types of Ca^2+stores may be established,maintained or refilled via distinct mechanisms.ER stores are refilled through sustained Ca^2+influx at ER-plasma membrane(PM)membrane contact sites(MCSs).In this review,we discuss the release and refilling mechanisms of intracellular small vesicular Ca^2+stores,with a special focus on lysosomes.Recent imaging studies of Ca2+release and organelle MCSs suggest that Ca^2+exchange may occur between two types of stores,such that the small stores acquire Ca^2+from the large stores via ER-vesicle MCSs.Hence vesicular stores like lysosomes may be viewed as secondary Ca^2+stores in the cell.展开更多
TRPP2 channel protein belongs to the superfamily of transient receptor potential(TRP) channels and is widely expressed in various tissues, including smooth muscle in digestive gut. Accumulating evidence has demonstrat...TRPP2 channel protein belongs to the superfamily of transient receptor potential(TRP) channels and is widely expressed in various tissues, including smooth muscle in digestive gut. Accumulating evidence has demonstrated that TRPP2 can mediate Ca^(2+) release from Ca^(2+) stores. However, the functional role of TRPP2 in gallbladder smooth muscle contraction still remains unclear. In this study, we used Ca^(2+) imaging and tension measurements to test agonist-induced intracellular Ca^(2+) concentration increase and smooth muscle contraction of guinea pig gallbladder, respectively. When TRPP2 protein was knocked down in gallbladder muscle strips from guinea pig, carbachol(CCh)-evoked Ca^(2+) release and extracellular Ca^(2+) influx were reduced significantly, and gallbladder contractions induced by endothelin 1 and cholecystokinin were suppressed markedly as well. CCh-induced gallbladder contraction was markedly suppressed by pretreatment with U73122, which inhibits phospholipase C to terminate inositol 1,4,5-trisphosphate receptor(IP3) production, and 2-aminoethoxydiphenyl borate(2APB), which inhibits IP3 recepor(IP3R) to abolish IP3R-mediated Ca^(2+) release. To confirm the role of Ca^(2+) release in CCh-induced gallbladder contraction, we used thapsigargin(TG)-to deplete Ca^(2+) stores via inhibiting sarco/endoplasmic reticulum Ca^(2+)-ATPase and eliminate the role of store-operated Ca^(2+) entry on the CCh-induced gallbladder contraction. Preincubation with 2 μmol L^(-1) TG significantly decreased the CCh-induced gallbladder contraction. In addition, pretreatments with U73122, 2APB or TG abolished the difference of the CCh-induced gallbladder contraction between TRPP2 knockdown and control groups. We conclude that TRPP2 mediates Ca^(2+) release from intracellular Ca^(2+) stores, and has an essential role in agonist-induced gallbladder muscle contraction.展开更多
文摘Cells utilize calcium ions(Ca^2+)to signal almost all aspects of cellular life,ranging from cell proliferation to cell death,in a spatially and temporally regulated manner.A key aspect of this regulation is the compartmen-talization of Ca^2+in various cytoplasmic organelles that act as intracellular Ca^2+stores.Whereas Ca^2+release from the large-volume Ca^2+stores,such as the endoplasmic reticulum(ER)and Golgi apparatus,are preferred for signal transduction,Ca^2+release from the small-volume individual vesicular stores that are dispersed throughout the cell,such as lysosomes,may be more useful in local regulation,such as membrane fusion and individualized vesicular movements.Conceivably,these two types of Ca^2+stores may be established,maintained or refilled via distinct mechanisms.ER stores are refilled through sustained Ca^2+influx at ER-plasma membrane(PM)membrane contact sites(MCSs).In this review,we discuss the release and refilling mechanisms of intracellular small vesicular Ca^2+stores,with a special focus on lysosomes.Recent imaging studies of Ca2+release and organelle MCSs suggest that Ca^2+exchange may occur between two types of stores,such that the small stores acquire Ca^2+from the large stores via ER-vesicle MCSs.Hence vesicular stores like lysosomes may be viewed as secondary Ca^2+stores in the cell.
基金supported by Anhui Provincial Natural Science Foundation (1208085MH181, 1108085J11)National Natural Science Foundation of China (81371284)Young Prominent Investigator Supporting Program from Anhui Medical University and National Training Program of Innovation and Entrepreneurship for Undergraduates (201310366012)
文摘TRPP2 channel protein belongs to the superfamily of transient receptor potential(TRP) channels and is widely expressed in various tissues, including smooth muscle in digestive gut. Accumulating evidence has demonstrated that TRPP2 can mediate Ca^(2+) release from Ca^(2+) stores. However, the functional role of TRPP2 in gallbladder smooth muscle contraction still remains unclear. In this study, we used Ca^(2+) imaging and tension measurements to test agonist-induced intracellular Ca^(2+) concentration increase and smooth muscle contraction of guinea pig gallbladder, respectively. When TRPP2 protein was knocked down in gallbladder muscle strips from guinea pig, carbachol(CCh)-evoked Ca^(2+) release and extracellular Ca^(2+) influx were reduced significantly, and gallbladder contractions induced by endothelin 1 and cholecystokinin were suppressed markedly as well. CCh-induced gallbladder contraction was markedly suppressed by pretreatment with U73122, which inhibits phospholipase C to terminate inositol 1,4,5-trisphosphate receptor(IP3) production, and 2-aminoethoxydiphenyl borate(2APB), which inhibits IP3 recepor(IP3R) to abolish IP3R-mediated Ca^(2+) release. To confirm the role of Ca^(2+) release in CCh-induced gallbladder contraction, we used thapsigargin(TG)-to deplete Ca^(2+) stores via inhibiting sarco/endoplasmic reticulum Ca^(2+)-ATPase and eliminate the role of store-operated Ca^(2+) entry on the CCh-induced gallbladder contraction. Preincubation with 2 μmol L^(-1) TG significantly decreased the CCh-induced gallbladder contraction. In addition, pretreatments with U73122, 2APB or TG abolished the difference of the CCh-induced gallbladder contraction between TRPP2 knockdown and control groups. We conclude that TRPP2 mediates Ca^(2+) release from intracellular Ca^(2+) stores, and has an essential role in agonist-induced gallbladder muscle contraction.
