MICROFILAMENTS DURING SEA-URCHIN FERTILIZATION - FLUORESCENCE DETECTION WITH RHODAMINYL PHALLOIDIN
MICROFILAMENTS DURING SEA-URCHIN FERTILIZATION - FLUORESCENCE DETECTION WITH RHODAMINYL PHALLOIDIN
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DOI：
10.1002/mrd.1120140402
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发表时间：
1986-01-01
期刊：
影响因子：
--
通讯作者：
SCHATTEN, G
中科院分区：
文献类型：
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作者：
CLINE, CA;SCHATTEN, G
用罗丹明标记的鬼笔环肽研究了海胆Arbacia punctulata和Lytechinus variegatus卵在受精和早期发育过程中微丝的分布。未受精卵的表面有许多点状荧光位点，罗丹明鬼笔环肽与之结合，表明存在肌动蛋白寡聚体或多聚体。在受精过程中，这种点状荧光模式开始改变。在受精后的30秒内，受精锥首先可以用这种技术检测到，它是卵子表面的直立结构。受精锥在8-9分钟时长到最大尺寸，并在受精后16分钟被吸收。受精后10分钟，受精卵的表面显示出许多荧光纤维，而不是在未受精卵中观察到的点状荧光，这表明微丝组装的破裂导致微绒毛伸长。延长的微丝在胞质分裂中持续存在。染色也可以在未受精、受精和卵裂卵的皮层中检测到。细胞松弛素E（10 μ M，30分钟）防止微丝伸长和胞质分裂，并降低受精后的皮质染色强度。在卵裂时，收缩环，出现狭窄的赤道纤维束，已被检测到在Lytechinus variegatus作为瞬态结构。
Rhodaminyl-labeled phalloidin is used to demonstrate the distribution of microfilaments during fertilization and early development in eggs of the sea urchins Arbacia punctulata and Lytechinus variegatus. The surface of unfertilized eggs have numerous punctate fluorescence sites at which rhodaminyl phalloidin binds, indicating the presence of actin oligomers or polymers. During fertilization this punctate pattern of fluorescence begins to change. Within thirty seconds of insemination, the fertilization cone is first detectable with this technique as an erect structure on the surface of the egg. The fertilization cone grows to a maximum size by 8-9 minutes, and is resorbed by 16 minutes after insemination. The surface of the fertilized egg displays numerous fluorescent fibers by 10 minutes after insemination rather than the punctate fluorescence observed in unfertilized eggs, indicative of the burst of microfilament assembly resulting in microvillar elongation. The elongated microfilaments persist through cytokinesis. Staining is also detected throughout the cortices of unfertilized, fertilized, and cleaving eggs. Cytochalasin E (10 .mu.M, 30 min) prevents microfilament elongation and cytokinesis and reduces the cortical staining intensity after fertilization. At cleavage, contractile rings, appearing as narrow equatorial bundles of fibers, have been detected in Lytechinus variegatus as transient structures.