钙稳态在体内集体迁移系统中的功能分析

论文摘要

集体细胞迁移（collective cell migration）在多细胞有机体的不同发育阶段的众多形态发生事件中都起到很重要的作用。而尽管钙稳态（Ca2+ homeostasis）在体外单细胞迁移中的作用和机制已经比较清楚,但是钙稳态如何调控和组织集体细胞迁移还知之甚少,尤其是在体内系统中。这里我们利用果蝇卵巢中的边界细胞团（border cell cluster）系统,发现在卵巢发生的第8期开始,当边界细胞的命运由静止向动态转化的过程中,钙的活性特异性上调。通过遗传筛选,我们发现胞浆中的钙稳态对于边界细胞的迁移至关重要,而线粒体钙稳态不是必须的。我们还发现胞浆钙稳态的维持依赖于钙离子诱导的钙离子释放（Ca2+-induced Ca2+release）,而非钙池操纵性钙内流（store-operated Ca2+ entry）。除了细胞内钙稳态,我们还发现由间隙连接介导的细胞间钙稳态在边界细胞的协同性方面扮演了重要角色。间隙连接蛋白inx2缺失的边界细胞表现出细胞非自主性的缺陷,最严重的情况下,边界细胞团甚至无法形成。意外的是,钙调蛋白（calmodulin）,钙调神经磷酸酶（calcineurin）以及果蝇中的经典蛋白激酶C（Pkc53E）,这三个非常重要的钙离子结合蛋白对于边界细胞的迁移都不是必须的,这暗示了经典的钙信号通路并非胞浆钙稳态向下传递信号的介导者。

论文目录

• Abstract
• 中文摘要
• Abbreviations
• Chapter Ⅰ: Review
• 2+ homeostasis and its maintenance'>  1 Ca²⁺ homeostasis and its maintenance
• 2+ shuttle system'>    1.1 Ca²⁺ shuttle system
• 2+ store'>      1.1.1 ER, the largest intracellular Ca²⁺store
•       1.1.2 Mitochondria, energy factory
• 2+ stores'>      1.1.3 Other Ca²⁺stores
• 2+ buffer system'>    1.2 Ca²⁺ buffer system
• 2+ buffer'>      1.2.1 ER Ca²⁺buffer
• 2+ buffer'>      1.2.2 Mitochondrial Ca²⁺buffer
• 2+ buffer'>      1.2.3 GA Ca²⁺buffer
• 2+ humeostasis in regulating cell migration'>  2 Ca²⁺ humeostasis in regulating cell migration
• 2+ homeostasis in cell migration'>    2.1 Cytosolic Ca²⁺ homeostasis in cell migration
• 2+ homeostasis in cell migration'>    2.2 ER Ca²⁺ homeostasis in cell migration
• 2+ homeostasis in cell migration'>    2.3 Mitochondrial Ca²⁺ homeostasis in cell migration
•   3 Genetic methods in Drosophila
•     3.1 GAL4/UAS system
•     3.2 FLP/FRT system
•   4 Drosophila oogenesis
•     4.1 Brief introduction
•     4.2 Border cell migration
•       4.2.1 Ecdysone signaling
•       4.2.2 JAK/STAT pathway
•       4.2.3 RTK cascade
•       4.2.4 Other regulatory components
• Introduction
• 2+ pattern and activity during Drosophila oogenesis'>Chapter Ⅱ: Specific Ca²⁺ pattern and activity during Drosophila oogenesis
• 2+ activity is higher in the follicle cells just adjacent to the polar cells andborder cells'>  1 Ca²⁺ activity is higher in the follicle cells just adjacent to the polar cells andborder cells
• 2+ dynamics during Drosophila oogenesis'>  2 Different forms of Ca²⁺ dynamics during Drosophila oogenesis
• 2+ homeostasis is required for border cell migration'>Chapter Ⅲ: Cytoplasmic Ca²⁺ homeostasis is required for border cell migration
•   1 CICR is required for border cell migration
•   2 Knockdown SERCA does not induce ER stress
• 2+ homeostasis are not required for border cellmigration'>  3 SOCE and mitochondrial Ca²⁺ homeostasis are not required for border cellmigration
• 2+ homeostasis is required for border cell migration'>Chapter Ⅳ: Intercellular Ca²⁺ homeostasis is required for border cell migration
•   1 Gap junctions are required for border cell migration
• 2+ homeostasis is required for the balance of STAT signaling'>  2 Intercellular Ca²⁺ homeostasis is required for the balance of STAT signaling
• 2+ homeostasis is also required for follicle cell migration'>  3 Intercellular Ca²⁺ homeostasis is also required for follicle cell migration
• 2+ signaling is not required for border cell migration'>Chapter Ⅴ: Canonical Ca²⁺ signaling is not required for border cell migration
• 2+ signaling pathways are not the mediators downstream of thecytoplasmic Ca²⁺ homeostasis'>  1 Canonical Ca²⁺ signaling pathways are not the mediators downstream of thecytoplasmic Ca²⁺homeostasis
• Discussion
• Reference
• Appendix
•   1 Fly genetics
•   2 Materials and methods
•     2.1 Live imaging of egg chambers
•     2.2 Immunostaining of egg chambers
•     2.3 Statistics and analysis of border cell migration index
• Publication
• Acknowledgement

文章来源

类型: 博士论文

作者: 郭炫

导师: 陈炯

关键词: 钙稳态,边界细胞,集体细胞迁移,果蝇

来源: 南京大学

年度: 2019

分类: 基础科学

专业: 生物学

单位: 南京大学

分类号: Q25

DOI: 10.27235/d.cnki.gnjiu.2019.000091

总页数: 76

文件大小: 5035k

下载量: 6