Characterization of Ca2+ signaling in the external yolk syncytial layer(E-YSL)during the late blastu

• 作 者: Andrew L. Miller
• 机 构: DivisionofLifeScienceandStateKeyLaboratoryofMolecularNeuroscience,HKUST,ClearWaterBay,HongKong,PRCan
• 出 处: 第三届全国斑马鱼研究大会
• 发表日期: 2013年10期
• 关键词: Zebrafish Yolk Syncytial Layer Yolk Syncytial Nuclei Aequorin Gastrulation

　　Using the bioluminescent Ca2+ reporter,f-aequorin,in conjunction with a custom-designed luminescence imaging system,we have identified both fast,short-range(i.e.,traveling at ~50 μm/sec over a distance of ~80 μm)and slow,long-range(i.e.,traveling at ~5 μm/sec over a distance of ~230 μm)propagating Ca2+ waves that are generated in the E-YSL.Embryos were loaded with f-aequorin into the top of the yolk at the 128-cell stage in order to preferentially load the Ca2+ reporter into the E-YSL as it developed.These two classes of Ca2+ waves begin during the late blastula period,between doming and ~30% epiboly stages,and continue for ~90 min(i.e.,until ~shield stage).The majority of both classes of waves are generated in the dorsal quadrant of the embryo and move in both directions around the E-YSL.