Stowers Institute for Medical Research, Kansas City, MO 64110, USA.
Dev Biol. 2011 Oct 15;358(2):309-17. doi: 10.1016/j.ydbio.2011.08.004. Epub 2011 Aug 16.
Examining calcium dynamics within the neural crest (NC) has the potential to shed light on mechanisms that regulate complex cell migration and patterning events during embryogenesis. Unfortunately, typical calcium indicators are added to culture media or have low signal to noise after microinjection into tissue that severely limit analyses to cultured cells or superficial events. Here, we studied in vivo calcium dynamics during NC cell migration and patterning, using a genetically encoded calcium sensor, GCaMP3. We discovered that trunk NC cells displayed significantly more spontaneous calcium transients than cranial NC cells, and during cell aggregation versus cell migration events. Spontaneous calcium transients were more prevalent during NC cell aggregation into discrete sympathetic ganglia (SG). Blocking of N-cadherin activity in trunk NC cells near the presumptive SG led to a dramatic decrease in the frequency of spontaneous calcium transients. Detailed analysis and mathematical modeling of cell behaviors during SG formation showed NC cells aggregated into clusters after displaying a spontaneous calcium transient. This approach highlights the novel application of a genetically encoded calcium indicator to study subsets of cells during ventral events in embryogenesis.
研究神经嵴（NC）内的钙动力学有可能揭示调节胚胎发生过程中复杂细胞迁移和模式形成事件的机制。不幸的是，典型的钙指示剂被添加到培养基中，或者在微注射到组织后信号与噪声的比值很低，这严重限制了对培养细胞或浅层事件的分析。在这里，我们使用遗传编码的钙传感器 GCaMP3 研究了 NC 细胞迁移和模式形成过程中的体内钙动力学。我们发现，与颅神经嵴细胞相比，躯干神经嵴细胞显示出更多的自发钙瞬变，并且在细胞聚集与细胞迁移事件中也是如此。自发钙瞬变在 NC 细胞聚集形成离散的交感神经节（SG）时更为普遍。阻断躯干神经嵴细胞中接近假定的 SG 的 N-钙粘蛋白活性会导致自发钙瞬变的频率急剧下降。对 SG 形成过程中细胞行为的详细分析和数学建模表明，NC 细胞在显示自发钙瞬变后聚集形成簇。这种方法突出了遗传编码钙指示剂在研究胚胎发生过程中腹侧事件中细胞亚群的新应用。
