Tetteroo P A, Bluemink J G, Dictus W J, van Zoelen E J, de Laat S W
Dev Biol. 1984 Jul;104(1):210-8. doi: 10.1016/0012-1606(84)90049-6.
The lateral mobility of plasma membrane lipids was analyzed during first cleavage of Xenopus laevis eggs by fluorescence photobleaching recovery (FPR) measurements, using the lipid analogs 5-(N-hexadecanoyl)aminofluorescein ("HEDAF") and 5-(N-tetradecanoyl)aminofluorescein ("TEDAF") as probes. The preexisting plasma membrane of the animal side showed an inhomogeneous, dotted fluorescence pattern after labeling and the lateral mobility of both probes used was below the detection limits of the FPR method (D much less than 10(-10) cm2/sec). In contrast, the preexisting plasma membrane of the vegetal side exhibited homogeneous fluorescence and the lateral diffusion coefficient of both probes used was relatively high (HEDAF, D = 2.8 X 10(-8) cm2/sec; TEDAF, D = 2.4 X 10(-8) cm2/sec). In the cleaving egg visible transfer of HEDAF or TEDAF from prelabeled plasma membrane to the new membrane in the furrow did not occur, even on the vegetal side. Upon labeling during cleavage, however, the new membrane was uniformly labeled and both probes were mobile, as in the vegetal preexisting plasma membrane. These data show that the membrane of the dividing Xenopus egg comprises three macrodomains: (i) the animal preexisting plasma membrane; (ii) the vegetal preexisting plasma membrane; (iii) the new furrow membrane.
利用脂质类似物5-(N-十六烷酰基)氨基荧光素(“HEDAF”)和5-(N-十四烷酰基)氨基荧光素(“TEDAF”)作为探针,通过荧光漂白恢复(FPR)测量分析了非洲爪蟾卵第一次卵裂期间质膜脂质的侧向流动性。动物侧预先存在的质膜在标记后呈现出不均匀的点状荧光模式,并且所使用的两种探针的侧向流动性均低于FPR方法的检测限(扩散系数D远小于10-10平方厘米/秒)。相比之下,植物侧预先存在的质膜呈现均匀荧光,并且所使用的两种探针的侧向扩散系数相对较高(HEDAF,D = 2.8×10-8平方厘米/秒;TEDAF,D = 2.4×10-8平方厘米/秒)。在正在分裂的卵中,即使在植物侧,也未观察到HEDAF或TEDAF从预先标记的质膜向沟中的新膜的可见转移。然而,在卵裂期间进行标记时,新膜被均匀标记,并且两种探针都是可移动的,这与植物侧预先存在的质膜情况相同。这些数据表明,正在分裂的非洲爪蟾卵的膜包括三个大区域:(i)动物侧预先存在的质膜;(ii)植物侧预先存在的质膜;(iii)新的沟膜。