Yazaki Ikuko, Uemura Isao
Department of Biology, Faculty of Science, Tokyo Metropolitan University, Fukazawa 2-1-1, Setagaya, 158, Tokyo, Japan.
Rouxs Arch Dev Biol. 1989 Oct;198(3):179-184. doi: 10.1007/BF02438943.
The blastomeres of sea urchin embryos have two surface regions with different properties. Numerous microvilli are present in the apical surface region, while the baso-lateral surface region, either on adjoining adjacent cells or facing the blastocoel, is smooth. When blastomeres are isolated from embryos and stained with fluorescein-isothiocyanate-labelled anti-(egg surface) antibody (anti-ES) prepared against membranes isolated from fertilized eggs, the apical microvillous region fluoresces while the smooth region does not [Yazaki I (1984) Acta Embryol Morphol Exp 5∶3-22]. In order to study quantitatively the 'bindability' of the membrane in the two regions to anti-ES, immunoelectron microscopy was used. Blastomeres isolated from embryos ofHemicentrotus pulcherrimus at the eight-cell stage were treated with rabbit anti-ES serum or pre-immune serum and then with ferritin-conjugated goat anti-(rabbit IgG) for 10 min at 0°C, mainly before fixation. About 10 times (maximally 45 times) more ferritin particles were counted per contour length in the microvillous surface region than in the smooth surface region.These results suggest that the membrane of the blastomeres of sea urchin embryos is a mosaic of two different membrane territories: one represented by the microvillous surface originating from the unfertilized egg, which binds anti-ES, the other by the smooth surface newly organized after the first cleavage, which does not react with anti-ES. The mechanism of segregation of the membrane into these two regions is discussed.
海胆胚胎的卵裂球有两个具有不同特性的表面区域。顶端表面区域有许多微绒毛,而基底外侧表面区域,无论是在相邻细胞上还是面对囊胚腔,都是光滑的。当从胚胎中分离出卵裂球并用针对从受精卵中分离出的膜制备的异硫氰酸荧光素标记的抗(卵表面)抗体(抗ES)进行染色时,顶端微绒毛区域会发出荧光,而光滑区域则不会[矢崎一(1984年)《实验胚胎形态学学报》5∶3 - 22]。为了定量研究这两个区域的膜与抗ES的“结合能力”,使用了免疫电子显微镜。从八细胞期的马粪海胆胚胎中分离出的卵裂球,在0°C下用兔抗ES血清或免疫前血清处理,然后用铁蛋白偶联的山羊抗(兔IgG)处理10分钟,主要是在固定之前。在微绒毛表面区域,每轮廓长度计数的铁蛋白颗粒比光滑表面区域多约10倍(最多45倍)。这些结果表明,海胆胚胎卵裂球的膜是由两个不同膜区域组成的镶嵌体:一个由源自未受精卵的微绒毛表面代表,它能结合抗ES,另一个由第一次卵裂后新形成的光滑表面代表,它不与抗ES反应。文中讨论了膜分离成这两个区域的机制。