TUMOR-NECROSIS-FACTOR-ALPHA GENE-EXPRESSION IN MOUSE OOCYTES AND FOLLICULAR CELLS
TUMOR-NECROSIS-FACTOR-ALPHA GENE-EXPRESSION IN MOUSE OOCYTES AND FOLLICULAR CELLS
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DOI：
10.1095/biolreprod48.4.707
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发表时间：
1993-04-01
影响因子：
3.6
通讯作者：
TERRANOVA, PF
中科院分区：
文献类型：
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作者：
CHEN, HL;MARCINKIEWICZ, JL;TERRANOVA, PF
通过原位杂交和免疫细胞化学，在成年周期小鼠中鉴定了转录和翻译肿瘤坏死因子α（TNF α）基因的卵巢细胞。两层或多层颗粒细胞的健康卵泡卵母细胞中TNF α mRNA的表达率>97%，蛋白质的表达率约为53%，而原始卵泡和单层颗粒细胞的卵母细胞中均未检测到TNF α mRNA和蛋白质。在早期闭锁卵泡中，只有13%含有TNF α蛋白，40%含有TNF α mRNA。在闭锁的晚期，在所有的卵母细胞中观察到强烈的免疫反应性TNF α，但TNF α mRNA仅存在于13%。在大约85%的卵泡中，卵泡膜和/或颗粒细胞显示TNF α mRNA杂交信号。间质内的巨噬细胞样细胞对TNF α mRNA和蛋白呈阳性。在黄体中，黄体细胞和巨噬细胞样细胞含有TNF α信息，而只有后者谱系含有免疫反应性TNF α。杂交信号和免疫反应性更强烈，在旧的黄体比本周期的黄体。北方印迹分析显示卵巢中的2.2 kb TNF α mRNA在周期中相对于28 S rRNA（组成型RNA）没有变化。类似地，TNF α杂交信号和免疫反应性在整个周期中似乎没有变化。这些结果表明，卵母细胞中TNF α基因的转录与免疫反应性TNF α的合成相一致，这些复杂的生化过程发生在小鼠卵泡发育的不同阶段。第二层颗粒细胞的形成与卵母细胞中TNF α基因转录的初始阶段相关联，因为TNF α mRNA和蛋白质在此阶段出现。我们的研究结果还表明，TNF α可能参与小鼠卵泡发育，闭锁和黄体功能的各个阶段。
ovarian cells that transcribe and translate the gene for tumor necrosis factor alpha (TNFalpha) were identified in the adult cyclic mouse by using in situ hybridization and immunocytochemistry. TNFalpha mRNA was observed in >97% and protein was contained in -53% of the oocytes of healthy follicles with two or more layers of granulosa cells, but neither was detectable in oocytes of primordial follicles and follicles with a single layer of granulosa cells. In early atretic follicles, only 13% contained TNFalpha protein and 40% contained TNFalpha mRNA. In late stages of atresia, intense immunoreactive TNFalpha was observed in all of the oocytes, but TNFalpha mRNA was present in only 13%. In approximately 85% of follicles, theca and/or granulosa cells exhibited TNFalpha mRNA hybridization signals. Macrophage-like cells within the interstitium were positive for TNFalpha mRNA and protein. In corpora lutea, luteal cells and macrophage-like cells contained TNFalpha message, while only the latter lineage contained immunoreactive TNFalpha. Hybridization signals and immunoreactivity were more intense in older corpora lutea than in corpora lutea of the present cycle. Northern blot analysis revealed a 2.2-kb TNFalpha mRNA in the ovary that was unchanged relative to 28S rRNA (constitutive RNA) during the cycle. Similarly, TNFalpha hybridization signals and immunoreactivity did not appear to change throughout the cycle. These results indicate that TNFalpha gene transcription in the oocyte coincides with the synthesis of immunoreactive TNFalpha and that these complex biochemical processes occur at distinct steps of follicular development in the mouse. The formation of the second layer of granulosa cells is coupled with the initial phase of TNFalpha gene transcription in the oocyte since TNFalpha MRNA and protein appear at this stage. Our findings also suggest that TNFalpha may be involved in various phases of follicular development, atresia, and luteal function in the mouse.