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Mechanisms of secretion studied with two-photon excitation imaging and uncaging

用双光子激发成像和解笼研究分泌机制

基本信息

批准号：
12144209
负责人：
NEMOTO Tomomi
金额：
$ 21.12万
依托单位：
National Institute for Physiological Sciences (2004)Okazaki National Research Institutes (2000-2003)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2004
项目状态：
已结题

项目摘要

Exocytosis and fluid-secretion in epithelial cells and synaptic functions were studied with two-photon excitation imaging and uncaging of caged-compounds.1) We have developed functional mapping of Ca^<2+>-activated channels using two-photon uncaging of caged-Ca^<2+> compound, which has an extraordinary spatial resolution because of the Ca^<2+> domain effect caused by caged-Ca^<2+> itself. In mouse pancreatic acinar preparations, we found that Ca^<2+>-activated Cl^- channel distributed both in the apical and basal membranes, but was absent in the lateral membrane. This distribution prevents the reuptake of Cl^- secreted into the lumen to the lateral membrane, and allows effective unidirectional transport of Cl^-from the basal membrane into the lumen. The lack of functional channels or transporters in the lateral membrane might be utilized in other epithelial transports, and supports the push-pull mechanism of fluid secretion in rodent pancreas.2) We have also developed a new imaging app … More roach to visualize exocytosis in intact tissue based on two-photon imaging of extracellular polar tracer (TEP imaging). TEP imaging revealed the sequential exocytosis in exocrine pancreas, where Ω-shaped profiles of individual vesicles were stably maintained by rapid coating of the vesicles with F-actin. Impairment of F-actin coating resulted in vacuole formation, which is an early sign of acute pancreatitis.3) We have found that sequential exocytosis also occurred in the guinea pig nasal gland, but with a substantial delay from the onset of fluid secretion.4) TEP imaging visualized exocytosis of individual insulin vesicles in the intact islets of Langerhans, and suggested that the fusion pore of insulin vesicles is made of lipid from its very beginning.5) We have developed an optical approach to stimulate neurons with submicron spatial resolution using two-photon uncaging of a caged-glutamate compound. We found that spine-head volumes tightly correlated with glutamate sensitivity, and enlargement of spine head underlay long-term potentiation. Less

我们用双光子激发成像和去笼化化合物研究了上皮细胞的胞吐和液体分泌以及突触功能。1)我们利用笼状化合物的双光子去中心化技术建立了钙激活通道的功能图谱，由于笼状化合物本身引起的钙域效应，它具有非凡的空间分辨率。在小鼠胰腺腺泡标本中，我们发现Ca~(2+)-激活的Cl~-通道在顶膜和基底膜上均有分布，但在侧膜上不存在。这种分布阻止了分泌到管腔内的氯离子重新摄取到侧膜，并允许从基膜到管腔的有效的单向运输。侧膜功能通道或转运体的缺乏可能被用于其他上皮转运，并支持啮齿动物胰腺液体分泌的推拉机制。2)我们还开发了一种新的成像APP…更多蟑螂，基于细胞外极性示踪剂的双光子成像(TEP成像)显示完整组织中的胞吐作用。TEP成像显示胰腺外分泌物中的连续胞吐，单个囊泡的Ω形状的轮廓通过快速包裹F-肌动蛋白而稳定地保持。F-肌动蛋白涂层受损导致空泡形成，这是急性胰腺炎的早期迹象。3)我们发现豚鼠鼻腺也发生了顺序的胞吐，但从液体分泌开始有很大的延迟。4)TEP成像显示了完整的朗格汉斯胰岛中单个胰岛素囊泡的胞吐，并提示胰岛素囊泡的融合孔从一开始就是由脂质组成的。5)我们已经开发出一种光学方法来刺激具有亚微米空间分辨率的神经元，方法是对笼养的谷氨酸化合物进行双光子浸泡。我们发现，脊柱头部的体积与谷氨酸敏感性密切相关，而脊柱头部的增大为长期增强奠定了基础。较少

项目成果

期刊论文数量（95）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Ion selectivities of the Ca2+ sensors for exocytosis in rat phaeochromocytoma cells

DOI：
10.1111/j.1469-7793.2001.t01-1-00627.x
发表时间：
2001-06
期刊：
The Journal of Physiology
影响因子：
0
作者：
T. Kishimoto;Ting‐ting Liu;Y. Ninomiya;Hiroshi Takagi;T. Yoshioka;G. Ellis‐Davies;Yasushi Miyashita;H. Kasai
通讯作者：
T. Kishimoto;Ting‐ting Liu;Y. Ninomiya;Hiroshi Takagi;T. Yoshioka;G. Ellis‐Davies;Yasushi Miyashita;H. Kasai

Kasai, H.: "Fast and cAMP-sensitive mode of Ca^<2+>-dependent exocytosis in pancreatic β cells"Diabetes. 51. S19-S24 (2002)

Kasai，H.：“胰腺β细胞中Ca 2+ 依赖性胞吐作用的快速和cAMP敏感模式”51.S19-S24(2002)。