Szego C M, Nazareno M B, Porter D D
J Cell Biol. 1977 May;73(2):354-65. doi: 10.1083/jcb.73.2.354.
The influence of estrogen on the subcellular localization and distribution of lysosomal components of preputial gland was investigated in the ovariectomized rat. Antisera of high titer and specificity toward high-density lysosomal lipoproteins of this organ were raised in rabbits. The immunologic effectiveness of the IgG fraction so obtained was confirmed by microcomplement fixation, immunodiffusion, and immunoelectrophoresis. By both direct and indirect immunofluorescence techniques, cryostat sections of preputial gland from the control animals exhibited pinpoint cytoplasmic fluorescence, of dimensions corresponding to those of lysosomes. In contrast, specific immunoreactive material in corresponding target cells from animals receiving 0.1 microng of estradiol-17 beta/100 g body wt only 2 min earlier was distributed more homogeneously, indicating release of antigen from the membrane-bounded organelles. Moreover, specific immunofluorescence became evident at cell surfaces and in peri- and supranuclear localization, sites essentially negative in the controls. These effects were intensified at 15 min, as well as by maximal physiologic dose (0.5 microng/100 g body wt) of hormone. The relatively less active epimer, estradiol-17 alpha, exhibited only very limited effectiveness by some of these criteria. These observations, taken together with independent biochemical and ultrastructural evidence, lead to the conclusion that structural labilization of lysosomal constituents and their translocation to the nuclear compartment are early correlates of estrogen action.
在去卵巢大鼠中研究了雌激素对包皮腺溶酶体成分亚细胞定位和分布的影响。用该器官的高密度溶酶体脂蛋白制备了高效价且特异性强的抗血清,将其注射到兔子体内。通过微量补体结合、免疫扩散和免疫电泳证实了所获得的IgG组分的免疫有效性。通过直接和间接免疫荧光技术,对照动物包皮腺的冰冻切片呈现出针尖状的细胞质荧光,其大小与溶酶体的大小相对应。相反,仅在2分钟前接受0.1微克17-β-雌二醇/100克体重的动物的相应靶细胞中的特异性免疫反应性物质分布更为均匀,表明抗原从膜结合细胞器中释放出来。此外,特异性免疫荧光在细胞表面以及核周和核上定位处变得明显,而在对照中这些部位基本呈阴性。在15分钟时以及给予最大生理剂量(0.5微克/100克体重)的激素时,这些效应会增强。相对活性较低的差向异构体17-α-雌二醇,根据其中一些标准仅表现出非常有限的作用。这些观察结果,结合独立的生化和超微结构证据,得出结论:溶酶体成分的结构不稳定及其向核区室的转运是雌激素作用的早期相关事件。