Horowitz S B
J Cell Biol. 1972 Sep;54(3):609-25. doi: 10.1083/jcb.54.3.609.
Ultralow temperature radioautography, suitable for the quantitative localization of diffusible solutes, was used to study the permeability of the nuclear envelope in the intact amphibian oocyte Sucrose-(3)H solutions were injected into mature oocytes, in volumes of 0 016-0 14% of that of the cell, and the subsequent movement of the solute was recorded. The resultant radioautographs show diffusion gradients in the cytoplasm and nucleus, and concentration gradients across the nuclear envelope Analysis of these gradients discloses that the nuclear envelope is as permeable as a comparable structure composed of cytoplasm, and is about 10(8) times more permeable than the oocyte plasma membrane The diffusion coefficient of sucrose in cytoplasm is 2 x 10(-6) cm(2)/sec, or about one-third its diffusivity in pure water. This reduction can probably be accounted for by an effective lengthening of the diffusional path because of obstruction by cytoplasmic inclusions. The nuclear: cytoplasmic sucrose concentration ratio at diffusional equilibrium is about 3 05, or 1.6 times as great as expected from the water content of the two compartments This asymmetry is attributed to an unavailability of 36% of the cytoplasmic water as solvent Finally, sucrose entry into oocytes from a bathing solution was monitored by whole cell analysis and radioautography. These and the microinjection results are consistent with a model in which sucrose entry into the cell is entirely limited by the permeability of the plasma membrane. The results are inconsistent with cell models that hypothesize a short-circuit transport route from the extracellular compartment to the nucleus, and with models in which cytoplasmic diffusion is viewed as limiting the rate of solute permeation.
超低温放射自显影技术适用于对可扩散溶质进行定量定位,被用于研究完整两栖类卵母细胞核膜的通透性。将含(3)H蔗糖溶液注入成熟卵母细胞,注射量为细胞体积的0.016 - 0.14%,随后记录溶质的后续移动情况。所得放射自显影片显示了细胞质和细胞核中的扩散梯度,以及跨核膜的浓度梯度。对这些梯度的分析表明,核膜的通透性与由细胞质组成的类似结构相当,并且比卵母细胞质膜的通透性高约10^8倍。蔗糖在细胞质中的扩散系数为2×10^(-6) cm²/秒,约为其在纯水中扩散系数的三分之一。这种降低可能是由于细胞质内含物的阻碍导致扩散路径有效延长所致。扩散平衡时核与细胞质中蔗糖浓度比约为3.05,是根据两个区室含水量预期值的1.6倍。这种不对称性归因于36%的细胞质水不可用作溶剂。最后,通过全细胞分析和放射自显影监测了蔗糖从浸泡溶液进入卵母细胞的情况。这些结果与显微注射结果一致,支持一个模型,即蔗糖进入细胞完全受质膜通透性的限制。这些结果与假设存在从细胞外区室到细胞核的短路运输途径的细胞模型以及认为细胞质扩散限制溶质渗透速率的模型不一致。