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. 2024:2818:171-177.
doi: 10.1007/978-1-0716-3906-1_11.

Visualization and Quantification of Rapid Chromosome Movements at Early Stages of Mouse Meiosis

Luciana Previato de Almeida  1 , Chih-Ying Lee  1 , Agustin Carbajal  1 , Rodrigo O de Castro  1 , Roberto J Pezza  2
Affiliations

Visualization and Quantification of Rapid Chromosome Movements at Early Stages of Mouse Meiosis

Luciana Previato de Almeida et al. Methods Mol Biol. 2024.

Abstract

Telomere-led rapid chromosome movements (RPMs) are a conserved characteristic of chromosome dynamics in meiosis. RPMs have been suggested to influence critical meiotic functions such as DNA repair and the association of the homologous chromosomes. Here, we describe a method using 3D time-lapse fluorescence imaging to monitor RPMs in Hoechst-stained mouse seminiferous tubules explants. We supplement visualization with customized quantitative motion analysis and in silico simulation. The ability to carry out live imaging, combined with quantitative image analysis, offers a sensitive tool to investigate the regulation of RPMs, chromosome reorganizations that precede dynamic mid-prophase events, and their contribution to faithful transmission of genetic information.

Keywords: Chromosome dynamics; Microscopy; Mouse gametogenesis; Spermatocyte; Telomere-led rapid prophase movements.

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Figures

Figure 1.
Figure 1.
Images show sample preparation and mounting before visualization. A. Use of culture dish for short term RPM measurements. B. sample preparation in a Ibidi microfluidics chamber. C. Ibidi microfluidic system and inverted epifluorescent microscope used to visualize RPMs in long term movies.
Figure 2.
Figure 2.
Maximum intensity projections of time lapse images of wild type zygotene spermatocytes. Red, blue, green, and yellow dots mark a defined centromere blob revealed by Hoechst 33342. Magnification bar represents 1 μm. The right panel show the traces of spots marked in each time frame.
Figure 3.
Figure 3.
Analysis of chromosomes movements using the software OMRFQANT. A. Example of an image opened with OMRFQANT. B. Single cell scoring. C. Splines animation of four spots (red, blue, green and yellow) scored in one cell. D. Time-lapse analysis window. E. Example of the final results obtained as a text file.
See this image and copyright information in PMC

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