Abstract

During cell division, mitotic chromosomes assemble and are equally distributed into two new daughter cells. The chromosome organisation of the two chromatids is essential for even distribution of genetic materials. Although the 11-nm fibre or nucleosome structure is well-understood as a fundamental fibrous structure of chromosomes, the reports on organisation of 30-nm basic chromatin fibres have been controversial, with debates on the contribution of 30-nm or thicker fibres to the higher order inner structure of chromosomes. Here, we used focused ion beam/scanning electron microscopy (FIB/SEM) to show that both 11-nm and 30-nm fibres are present in the human metaphase chromosome, although the higher-order periodical structure could not be detected under the conditions employed. We directly dissected the chromosome every 10-nm and observed 224 cross-section SEM images. We demonstrated that the chromosome consisted of chromatin fibres of an average diameter of 16.9-nm. The majority of the chromatin fibres had diameters between 5 and 25-nm, while those with 30-nm were in the minority. The reduced packaging ratio of the chromatin fibres was detected at axial regions of each chromatid. Our results provide a strong basis for further discussions on the chromosome higher-order structure.

Highlights

  • IntroductionMitotic chromosomes assemble and are distributed into two new daughter cells

  • During cell division, mitotic chromosomes assemble and are distributed into two new daughter cells

  • Using ChromEM tomography, 5–24-nm chromatin fibres were recently detected[12]. To clarify these inconsistencies and to elucidate the presence of higher-order fibrous structure of chromatin, including the 30-nm fibre, which would be essential for the modelling of the chromosome structure, we used focused ion beam/scanning electron microscopy (FIB/SEM)

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Summary

Introduction

Mitotic chromosomes assemble and are distributed into two new daughter cells. We used focused ion beam/scanning electron microscopy (FIB/SEM) to show that both 11-nm and 30-nm fibres are present in the human metaphase chromosome, the higher-order periodical structure could not be detected under the conditions employed. Because 30-nm fibrous structures have been repeatedly observed under scanning electron microscopy (SEM) at the surface of metaphase chromosomes[3], they have been referred to as the basic chromatin fibre. Using ChromEM tomography, 5–24-nm chromatin fibres were recently detected[12] To clarify these inconsistencies and to elucidate the presence of higher-order fibrous structure of chromatin, including the 30-nm fibre, which would be essential for the modelling of the chromosome structure, we used focused ion beam/scanning electron microscopy (FIB/SEM).

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