Synaptotagmin VII (Syt VII), which has a higher Ca2+ affinity and slower disassembly kinetics with lipid than Syt I and Syt IX, was regarded as being uninvolved in synaptic vesicle (SV) exocytosis but instead possibly...Synaptotagmin VII (Syt VII), which has a higher Ca2+ affinity and slower disassembly kinetics with lipid than Syt I and Syt IX, was regarded as being uninvolved in synaptic vesicle (SV) exocytosis but instead possibly as a calcium sensor for the slower kinetic phase of dense core vesicles (DCVs) release. By using high temporal resolution capacitance and amperometry measurements, it was demonstrated that the knockdown of endogenous Syt VII attenuated the fusion of DCV with the plasma membrane, reduced the amplitude of the exocytotic burst of the Ca2+-triggered DCV release without affecting the slope of the sustained component, and blocked the fusion pore expansion. This suggests that Syt VII is the Ca2+ sensor of DCV fusion machinery and is an essential factor for the establishment and maintenance of the pool size of releasable DCVs in PC12 cells.展开更多
A number of transmembrane receptors are targeted to the nucleus and convincingly localized therein. However, what remains a conundrum is how these cell-surface receptors end up in the nucleus. In this study, we report...A number of transmembrane receptors are targeted to the nucleus and convincingly localized therein. However, what remains a conundrum is how these cell-surface receptors end up in the nucleus. In this study, we reported that the transmembrane receptor phosphorylated TrkA was located in a series of carrier vesicles, including ring-like vesicles near the plasma membrane, large core vesicles and small dense core vesicles around the nuclei, as well as in the nucleus in human glioma cell line U251 using immunocytochemistry and immunofluorescence staining. Meanwhile, we also showed that small dense core vesicles budded from large core vesicles, and interacted with the nuclear envelope. Accordingly, our results suggested that such a series of membrane compartments might be involved in the pathway of nuclear translocation of the transmembrane receptor TrkA.展开更多
The molecular mechanisms by which dense core vesicles(DCVs) translocate,tether,dock and prime are poorly understood.In this study,Caenorhabditis elegans was used as a model organism to study the function of Rab protei...The molecular mechanisms by which dense core vesicles(DCVs) translocate,tether,dock and prime are poorly understood.In this study,Caenorhabditis elegans was used as a model organism to study the function of Rab proteins and their effectors in DCV exocytosis.RAB-27/AEX-6,but not RAB-3,was found to be required for peptide release from neurons.By analyzing the movement of DCVs approaching the plasma membrane using total internal reflection fluorescence microscopy,we demonstrated that RAB-27/AEX-6 is involved in the tethering of DCVs and that its effector rabphilin/RBF-1 is required for the initial tethering and subsequent stabilization by docking.展开更多
基金Supported by the National Natural Science Foundation of China (Grant Nos. 30500117 and 30670502)
文摘Synaptotagmin VII (Syt VII), which has a higher Ca2+ affinity and slower disassembly kinetics with lipid than Syt I and Syt IX, was regarded as being uninvolved in synaptic vesicle (SV) exocytosis but instead possibly as a calcium sensor for the slower kinetic phase of dense core vesicles (DCVs) release. By using high temporal resolution capacitance and amperometry measurements, it was demonstrated that the knockdown of endogenous Syt VII attenuated the fusion of DCV with the plasma membrane, reduced the amplitude of the exocytotic burst of the Ca2+-triggered DCV release without affecting the slope of the sustained component, and blocked the fusion pore expansion. This suggests that Syt VII is the Ca2+ sensor of DCV fusion machinery and is an essential factor for the establishment and maintenance of the pool size of releasable DCVs in PC12 cells.
基金the National Natural Science Foundation of China (Grant Nos. 30270639, 30570894 and 30570981)the Doctoral Research Foundation of the Jiangsu University, China (Grant No. 02JDG028)
文摘A number of transmembrane receptors are targeted to the nucleus and convincingly localized therein. However, what remains a conundrum is how these cell-surface receptors end up in the nucleus. In this study, we reported that the transmembrane receptor phosphorylated TrkA was located in a series of carrier vesicles, including ring-like vesicles near the plasma membrane, large core vesicles and small dense core vesicles around the nuclei, as well as in the nucleus in human glioma cell line U251 using immunocytochemistry and immunofluorescence staining. Meanwhile, we also showed that small dense core vesicles budded from large core vesicles, and interacted with the nuclear envelope. Accordingly, our results suggested that such a series of membrane compartments might be involved in the pathway of nuclear translocation of the transmembrane receptor TrkA.
基金supported by grants from National Basic Research Program of China(2010CB833701)The National Natural Science Foundation of China(30870564,90913022)Key Inndvative Project of Chinese Academy of Sciences(KSCX2-SW-224)~~
基金supported by the National Basic Research Program of China(Grant No. 2010CB833701)the National Natural Science Foundation of China(Grant Nos. 30870564 and 90913022)the CAS Project(Grant No.KSCX2-SW-224)
文摘The molecular mechanisms by which dense core vesicles(DCVs) translocate,tether,dock and prime are poorly understood.In this study,Caenorhabditis elegans was used as a model organism to study the function of Rab proteins and their effectors in DCV exocytosis.RAB-27/AEX-6,but not RAB-3,was found to be required for peptide release from neurons.By analyzing the movement of DCVs approaching the plasma membrane using total internal reflection fluorescence microscopy,we demonstrated that RAB-27/AEX-6 is involved in the tethering of DCVs and that its effector rabphilin/RBF-1 is required for the initial tethering and subsequent stabilization by docking.
