Live Cell Imaging of Male Meiosis in Arabidopsis by a Landmark-based System.

Maria Prusicki,Christian Fleck,Arp Schnittger,Emma Keizer,Rik Van Rosmalen

doi:10.21769/bioprotoc.3611

Maria Prusicki, Christian Fleck + Show 3 more

Open Access

https://doi.org/10.21769/bioprotoc.3611

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Abstract

Live cell imaging has tremendously promoted our understanding of cellular and subcellular processes such as cell division. Here, we present a step-by-step protocol for a robust and easy-to-use live cell imaging approach to study male meiosis in the plant Arabidopsis thaliana as recently established. Our method relies on the concomitant analysis of two reporter genes that highlight chromosome configurations and microtubule dynamics. In combination, these reporter genes allowed the discrimination of five cellular parameters: cell shape, microtubule array, nucleus position, nucleolus position, and chromatin condensation. These parameters can adopt different states, e.g., the nucleus position can be central or lateral. Analyzing how tightly these states are associated gives rise to landmark stages that in turn allow a quantitative and qualitative dissection of meiotic progression. We envision that such an approach can also provide valuable criteria for the analysis of cell differentiation processes outside of meiosis.

Highlights

[Background] Meiosis is a special cell division cycle that serves two major purposes
Research on plant meiosis has largely relied on the analysis of fixed cells, e.g., for chromosome spreads and immunolocalization studies, both for Arabidopsis (Armstrong et al, 2009; Martinez-Garcia and Pradillo, 2017; Parra-Nunez et al, 2020; Sims et al, 2020) as well as for other plant species (Chelysheva et al, 2013; Sepsi et al, 2018; Darrier et al, 2020; Stack et al, 2020)
These techniques have been and continue to be important tools as they offer a great spatial resolution to address the structure of meiotic chromosomes and the localization of meiotic regulators for instance demonstrated by an immuno-cytological analysis of the 3D configuration of meiotic chromosomes (Hurel et al, 2018)

Summary

Introduction

[Background] Meiosis is a special cell division cycle that serves two major purposes. Surface-sterilize seeds of Arabidopsis thaliana, carrying a fluorescent reporter expressed at meiotic stages (an example is available in Prusicki et al, 2019). If instead you are planning to store the sterilized seeds in the tube for some time, let the tubes open under a clean bench for a minimum of 30 min to ensure the gas has completely evaporated.

Results

Conclusion