以单细胞免疫荧光技术观察烟草细胞融合过程中微管骨架的变化

本文建立了单细胞免疫荧光标记技术并以此结合单对细胞融合技术对细胞融合过程中微管骨架组织形式的动态变化进行了追踪观察。发现在聚乙二醇(PEG)诱导条件下,一旦细胞开始粘连,细胞内微管骨架便开始解聚。在细胞融合的整个过程中一直维持着这种解聚的状态,直到融合完成,在后续的培养中微管骨架才重新出现。在微管骨架呈解聚状态时融合产物不能完成与另外的细胞融合。实验揭示了细胞的再融合能力可能受细胞本身微管骨架状态的影响。该结果为解释高等植物如何避免多精入卵提供了新的可能性。

THE CHANGE OF MICROTUBULE ORGANIZATION INFLUENCING CELL ABILITY FOR FUSION

It is well known that plants have ability to avoid polygamy in angiosperm, however, its mechanism is still poorly understood. In our previous work of in vitro fertilization we found that once an egg cell fused with a sperm cell, it could hardly fuse with other sperm within a cer- tain period. Therefore we proposed that fusing egg cell must suffer some intrinsic changes during the fusion, such as the change of microtubule organization, which may prevent the egg cell from another fusion with extra sperm cells. In present work, the dynamic change of microtubule organi- zation during the process of cell fusion in Nicotiana tabacum was observed through the single cell immunofluorescence microscopy. It was found that once cells began to adhere and fuse in Polyethylene Glycol (PEG), the microtubule immediately depolymerized. During this period of fusion and before microtubule polymerized again in the fusion product, the cell was unable to fuse with other cells. The results suggested microtubule organization was likely involved in the mechanisms of controlling cell fusion, and offered a clear indication that similar mechanism might also con- tribute to plant avoiding polygamy.