Reima I, Lehtonen E
Differentiation. 1985;30(1):68-75. doi: 10.1111/j.1432-0436.1985.tb00515.x.
Mouse oocytes, cleavage-stage embryos, and blastocyst-stage embryos were studied to show the distribution of both an immunoanalog to nonerythroid spectrin (p 230) and F-actin. Using antibodies to nonerythroid spectrin, diffuse, positive cytoplasmic fluorescence was regularly seen in oocytes and embryo cells. The presence of nonerythroid spectrin in oocytes was confirmed by immunoblotting. Oocytes usually exhibited an inconspicuous submembranous layer of nonerythroid spectrin, which was more pronounced in the area of the polar body. Oocytes regularly exhibited a peripheral concentration of actin. Throughout the cleavage and blastocyst stages, a cortical layer of nonerythroid spectrin and actin was usually observed in embryo cells. These submembranous layers on the outer surface of the embryo were relatively thin as compared to those in areas of intercellular contact. The contact areas regularly showed distinct positive staining, including a concentration of label at the most peripheral region of each contact area. This resulted in the presence of ring-like fluorescence around each blastomere. Nonerythroid spectrin and actin showed concentration to the contact area between the oocyte and the polar body. Although the general localization patterns of nonerythroid spectrin and actin were similar, double-staining experiments revealed that slightly different planes of focus were necessary to obtain sharp definition of the fluorescence of these components in areas of intercellular contact: the ring-like concentration of nonerythroid spectrin appeared to be localized more peripherally than that of actin. The cells of preimplantation embryos show motile features that include actual cell movements and striking changes in cell shape (e.g., during compaction). The submembraneous layers of nonerythroid spectrin and actin may contribute to the regulation of the deformability and thus the shape of embryo cells.(ABSTRACT TRUNCATED AT 250 WORDS)
研究了小鼠卵母细胞、分裂期胚胎和囊胚期胚胎,以显示非红细胞血影蛋白(p230)免疫类似物和F-肌动蛋白的分布。使用针对非红细胞血影蛋白的抗体,在卵母细胞和胚胎细胞中经常可见弥漫性的阳性细胞质荧光。通过免疫印迹证实了卵母细胞中存在非红细胞血影蛋白。卵母细胞通常表现出不明显的非红细胞血影蛋白膜下亚层,在极体区域更为明显。卵母细胞 regularly表现出肌动蛋白的外周聚集。在整个分裂期和囊胚期,胚胎细胞中通常观察到一层非红细胞血影蛋白和肌动蛋白的皮质层。与细胞间接触区域相比,胚胎外表面的这些膜下亚层相对较薄。接触区域 regularly显示出明显的阳性染色,包括每个接触区域最外周区域的标记聚集。这导致每个卵裂球周围出现环状荧光。非红细胞血影蛋白和肌动蛋白在卵母细胞和极体之间的接触区域聚集。尽管非红细胞血影蛋白和肌动蛋白的一般定位模式相似,但双重染色实验表明,在细胞间接触区域获得这些成分荧光的清晰定义需要略有不同的聚焦平面:非红细胞血影蛋白的环状聚集似乎比肌动蛋白更位于外周。植入前胚胎的细胞表现出运动特征,包括实际的细胞运动和细胞形状的显著变化(例如,在致密化过程中)。非红细胞血影蛋白和肌动蛋白的膜下亚层可能有助于调节胚胎细胞的可变形性,从而调节其形状。(摘要截断于250字)