González-García J M, Rufas J S, Antonio C, Suja J A
Unidad de Biología Celular, Departamento de Biología, Edificio de Ciencias Biológicas, Universidad Autónoma de Madrid, E-28049 Madrid, Spain.
Chromosoma. 1995 Dec;104(4):287-97. doi: 10.1007/BF00352260.
During early embryogenesis of the nematode Parascaris univalens (2n=2) the processes of chromatin diminution and segregation of the germ and somatic cell lineages take place simultaneously. In this study we analyzed the nucleolar cycle in early embryos, both in germinal and somatic blastomeres, by means of silver staining and antibodies against the nucleolar protein fibrillarin. We observed an identical nucleolar cycle in both types of blastomeres, hence, the chromatin diminution process has no effect on the nucleolar cycle of somatic blastomeres. We report the existence of outstanding differences between this cycle and those previously reported during early embryogenesis of other species. There is a true nucleolar cycle in early embryos that shows a peculiar nucleolar disorganization at prophase, and a preferential localization of prenucleolar bodies only on the euchromatic regions during nucleologenesis. Moreover, fibrillarin does not form a perichromosomal sheath in metaphase or anaphase holocentric chromosomes, probably owing to their special centromeric organization. The number and location of nucleolus organizer regions (NORs) in the chromosomal complement have been determined using silver impregnation, chromomycin A3/ distamycin A staining, and fluorescent in situ hybridization using an rDNA probe. There are only two NORs, one per chromosome, and these are located in the middle of the euchromatic central regions. This location implies that no rDNA sequences are lost in blastomeres after chromatin diminution. Moreover, the constant presence of two nucleoli in somatic blastomeres suggests that NORs are not affected during the fragmentation of euchromatic regions when this process occurs.
在单子宫副蛔虫(2n = 2)的早期胚胎发育过程中,染色质消减以及生殖细胞系和体细胞系的分离过程同时发生。在本研究中,我们通过银染以及针对核仁蛋白纤维蛋白原的抗体,分析了早期胚胎中生殖和体细胞卵裂球的核仁周期。我们在两种类型的卵裂球中观察到相同的核仁周期,因此,染色质消减过程对体细胞卵裂球的核仁周期没有影响。我们报告了这个周期与之前报道的其他物种早期胚胎发育过程中的周期存在显著差异。早期胚胎中存在一个真正的核仁周期,在前期显示出特殊的核仁解体,并且在核仁形成过程中,核仁前体仅优先定位于常染色质区域。此外,在中期或后期的全着丝粒染色体中,纤维蛋白原不会形成染色体周鞘,这可能是由于它们特殊的着丝粒组织。使用银染、放线菌素A3/偏端霉素A染色以及使用rDNA探针的荧光原位杂交,确定了染色体组中核仁组织区(NORs)的数量和位置。只有两个NORs,每条染色体一个,并且它们位于常染色质中央区域的中间。这个位置意味着在染色质消减后,卵裂球中没有rDNA序列丢失。此外,体细胞卵裂球中持续存在两个核仁,这表明当这个过程发生时,NORs在常染色质区域片段化过程中不受影响。