Ganion L R
Department of Physiology and Health Science, Ball State University, Muncie, Indiana 47306.
Anat Rec. 1991 Jun;230(2):218-24. doi: 10.1002/ar.1092300209.
The cytoplasmic distribution of poly(A)+ mRNA and its relationship to annulate lamellae were examined in developing Necturus maculosus oocytes by in situ hybridization with [3H]poly(U). The specificity of [3H]poly(U) binding was tested by incubating control ovarian sections with either KOH or RNase A before in situ hybridization. In both experiments, the silver grain densities were markedly reduced. Poly(A)+ RNA is uniformly distributed in the cytoplasm until the mid-growth phase and then later in vitellogenesis becomes localized in the subcortical ooplasm. The silver grain density in the cytoplasm varied during oogenesis and was greatest in previtellogenic oocytes. Annulate lamellae commonly are observed with the light microscope in oocytes prior to vitellogenesis. In such oocytes, the labeled mRNA probe is observed over cytoplasmic regions of annulate lamellae. The results suggests that a differential localization of messenger RNA occurs during oogenesis in Necturus maculosus. Furthermore, poly(A)+ RNA is present in cytoplasmic regions of annulate lamellae.
通过用[3H]多聚尿苷进行原位杂交,研究了正在发育的斑泥螈卵母细胞中多聚腺苷酸(poly(A)+)mRNA的细胞质分布及其与环孔片层的关系。在原位杂交之前,通过用氢氧化钾或核糖核酸酶A孵育对照卵巢切片来测试[3H]多聚尿苷结合的特异性。在这两个实验中,银颗粒密度均显著降低。在生长中期之前,多聚腺苷酸(poly(A)+)RNA均匀分布于细胞质中,随后在卵黄发生后期定位于皮质下卵质中。在卵子发生过程中,细胞质中的银颗粒密度有所变化,在前卵黄生成期的卵母细胞中密度最大。在光学显微镜下,通常在卵黄发生之前的卵母细胞中观察到环孔片层。在这类卵母细胞中,在环孔片层的细胞质区域观察到标记的mRNA探针。结果表明,在斑泥螈卵子发生过程中,信使RNA发生了差异性定位。此外,多聚腺苷酸(poly(A)+)RNA存在于环孔片层的细胞质区域。