Department of Animal Pathology, Faculty of Veterinary Sciences, University of Zaragoza, Zaragoza, Spain.
Int J Parasitol. 2010 Mar 15;40(4):453-62. doi: 10.1016/j.ijpara.2009.09.008. Epub 2009 Oct 29.
In Eimeria tenella, meiotic division occurs exclusively in oocysts within the first 8h of sporulation. Difficulties with the wall-oocyst breakage in gaining access to chromosomes during meiosis have resulted in a scarcity of morphological data on Eimeria chromosomes. This study tracks the general behaviour of telomeres, attachment plaques and synaptonemal complexes in the nucleus of the meiotic oocyst of E. tenella. Fluorescence microscopy methods, in combination with immunoelectron microscopy techniques, were applied to obtain a series of time-lapse images during oocyst sporulation. Antibodies to Structural Maintenance of Chromosome proteins SMC1 and SMC3, and lamin were labelled with either fluorescence or colloidal gold to visualise the telomeres, central elements of the synaptonemal complex (SC) and nuclear periphery, respectively, at both the structural and ultrastructural levels. Using oocyst spreads and ultrathin sections of fixed oocysts it was possible to study telomere dynamics at stages during meiosis. The stages of the meiotic prophase I are delineated on the basis of the telomere position and the SC synapsis and desynapsis. During the leptotene stage, at 4h following the start of sporulation, meiotic chromosomes attached to the nuclear envelope. At that stage, chromosome synapsis was initiated in the telomeric regions but no interstitial synapsis pairing was observed. In the zygotene stage, telomere signals were clustered in a limited area of the nuclear envelope. Bouquet formation occurred at 5h after the start of sporulation, whereas chromosomes did not appear completely synapsed until the pachytene stage at 6h of sporulation. Desynapsis was observed at 8h of sporulation during the diplotene stage. This study provides the first morphological description of both the behaviour of the chromosomes and the timing of the prophase I stages in the meiotic nucleus of E. tenella.
在柔嫩艾美耳球虫中,减数分裂仅发生在孢子化的前 8 小时内的卵囊内。由于在减数分裂过程中卵囊壁破裂以获得染色体存在困难,因此关于柔嫩艾美耳球虫染色体的形态学数据很少。本研究跟踪了柔嫩艾美耳球虫减数分裂卵囊核中端粒、附着斑和联会复合体的一般行为。荧光显微镜方法与免疫电镜技术相结合,用于在卵囊孢子发生过程中获得一系列延时图像。针对结构维持染色体蛋白 SMC1 和 SMC3 的抗体以及核纤层蛋白用荧光或胶体金标记,分别在结构和超微结构水平上可视化端粒、联会复合体的中心元件(SC)和核周界。使用卵囊铺片和固定卵囊的超薄切片,可以研究减数分裂过程中各阶段的端粒动力学。基于端粒位置和 SC 联会和去联会,划分了减数分裂前期 I 的阶段。在孢子发生开始后的 4 小时,即细线期,减数染色体附着在核膜上。在那个阶段,在端粒区域开始发生染色体联会,但没有观察到间质联会配对。在合线期,端粒信号聚集在核膜的有限区域。在孢子发生开始后 5 小时发生束状形成,而直到孢子发生 6 小时的粗线期,染色体才完全联会。在孢子发生的双线期的 8 小时观察到去联会。本研究首次提供了柔嫩艾美耳球虫减数分裂核中染色体行为和前期 I 阶段时间的形态描述。