Alvarez Marco, Quezada Claudia, Molina Alfredo, Krauskopf Manuel, Vera M Ines, Thiry Marc
Departamento de Ciencias Biológicas, Universidad Andrés Bello, Santiago, Chile.
Biol Cell. 2006 Aug;98(8):457-63. doi: 10.1042/BC20060006.
The eurythermal fish carp (Cyprinus carpio) adjusts to the seasonal changes in the temperature and photoperiod of its habitat through diverse cellular and molecular mechanisms. We have observed that ribosomal biogenesis is modulated during the acclimatization process and correlates with profound phenotypic changes, reflecting a seasonal-dependent ultrastructural appearance of the nucleolar components. Previous studies using classical techniques showed that in winter-adapted carp the nucleolus appears to be segregated. In the present work, we have reassessed the nucleolar ultrastructural organization of the carp in summer- and winter-adapted fish by using more specific cytochemical and immunocytological techniques.
The acetylation method provided evidence that the nucleolar organization is different between winter- and summer-adapted carp. In winter-adapted fish the fibrillar component appears as a unique mass surrounded by several granular caps, whereas in summer-adapted carp the fibrillar component forms few cordons surrounded by granular masses. The nucleolar structure and distribution of the condensed chromatin observed varies upon seasonal acclimatization. In winter the nucleolar chromatin is densely packed in masses that surround the nucleolus, whereas during summer it displays a rather looser organization formed by filaments that not only surround the nucleolus, but also go through the nucleolar body. Using the TdT (terminal deoxynucleotidyl transferase)-immunogold labelling technique, we detected condensed and decondensed nucleolar chromatin, and found some labelling of fibrillar components in both seasons. When liver tissue from summer-adapted carp was treated with AMD (actinomycin D), we observed that the rearrangement of the nucleolar components and condensed chromatin were similar to that found in winter-adapted fish, with differences in the distribution of the perinucleolar chromatin.
The acetylation and TdT-immunogold labelling experiments indicated that the rearrangement of the nucleolar components of winter-adapted carp is very similar to the AMD-treated summer-adapted carp nucleolus, with the latter representing the repression of the ribosomal biogenesis that occurs during the cold season. Nevertheless, the distribution of the condensed perinucleolar chromatin in winter-adapted carp compared with AMD-treated cells suggests that the transcription of rRNA genes in winter-adapted fish is less strongly inhibited and does not lead to the classical segregation of the nucleolus of that described after AMD treatment. In addition, we have confirmed that carp hepatocyte nucleoli comprise only two main structural compartments: a fibrillar component and a granular component. Fibrillar centres were not observed.
广温性鱼类鲤鱼(Cyprinus carpio)通过多种细胞和分子机制来适应其栖息地温度和光周期的季节性变化。我们观察到核糖体生物合成在适应过程中受到调节,并与深刻的表型变化相关,这反映了核仁成分的季节性超微结构外观。先前使用经典技术的研究表明,适应冬季的鲤鱼核仁似乎是分离的。在本研究中,我们通过使用更特异的细胞化学和免疫细胞化学技术,重新评估了适应夏季和冬季的鲤鱼的核仁超微结构组织。
乙酰化方法提供了证据,表明适应冬季和夏季的鲤鱼的核仁组织不同。在适应冬季的鱼类中,纤维成分表现为被几个颗粒帽包围的独特团块,而在适应夏季的鲤鱼中,纤维成分形成少数被颗粒团块包围的索带。观察到的浓缩染色质的核仁结构和分布随季节性适应而变化。在冬季,核仁染色质密集堆积在围绕核仁的团块中,而在夏季,它呈现出一种较为松散的组织形式,由不仅围绕核仁而且穿过核仁体的细丝组成。使用TdT(末端脱氧核苷酸转移酶)免疫金标记技术,我们检测到了浓缩和去浓缩的核仁染色质,并在两个季节中都发现了纤维成分的一些标记。当用放线菌素D(AMD)处理适应夏季的鲤鱼的肝脏组织时,我们观察到核仁成分和浓缩染色质的重排与适应冬季的鱼类相似,只是核仁周围染色质的分布有所不同。
乙酰化和TdT免疫金标记实验表明,适应冬季的鲤鱼的核仁成分重排与经AMD处理的适应夏季的鲤鱼核仁非常相似,后者代表了寒冷季节期间发生的核糖体生物合成的抑制。然而,与经AMD处理的细胞相比,适应冬季的鲤鱼中浓缩的核仁周围染色质的分布表明,适应冬季的鱼类中rRNA基因的转录受到的抑制较弱,并且不会导致AMD处理后所描述的核仁的经典分离。此外,我们已经证实鲤鱼肝细胞核仁仅由两个主要结构区室组成:纤维成分和颗粒成分。未观察到纤维中心。