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

Prusicki, Maria Ada; Keizer, Emma M; Rosmalen, Rik P. van; Fleck, Christian; Schnittger, Arp

doi:10.21769/bioprotoc.3611

Cited by 3 publications

(5 citation statements)

References 29 publications

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“…Different time points of fixation were carried out (12, 28, and 38 h). Given the known meiotic timecourse of A. thaliana [16][17][18] (Figure 3a), the different time points and inhibition methods could provide us with insights about at what stage of meiosis TOPII may be functional. The treatment with merbarone (Figure 3b-e) produced the presence of multiple ABs at AI (6% n = 50) (Figure 3b), chromosome fragmentation (7% n = 43) (Figure 3c), missegregation (4% n = 25) (Figure 3d), and micronuclei at telophase II (10% n = 40) (Figure 3e).…”

Section: Analysis Of Wt Male Meiosis Treated With Topii Inhibitorsmentioning

confidence: 99%

“…The meiotic time-course in A. thaliana has been previously characterized by using the nucleotide analogue BrdU [16,17]. This time course has been validated recently with life cell imaging techniques [18].…”

Section: Male Meiocytes Treated With Topii Inhibitors Show Anaphase B...mentioning

confidence: 99%

“…Merbarone (Sigma) was prepared at concentrations of 1 and 10 µM, etoposide (Tocris Bioscience) at 0.05 and 5 µM, and ICRF-187 (Sigma) at 0.1 and 100 µg/mL. Based on the 33 h meiotic time-course of A. thaliana [16][17][18], inhibitor treatments were applied to WT plants in a 2 h pulse or continuously. These two ways to apply the inhibitor were used for each drug and concentration.…”

Section: Treatments With Topii Inhibitorsmentioning

confidence: 99%

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The Role of DNA Topoisomerase Binding Protein 1 (TopBP1) in Genome Stability in Arabidopsis

Parra-Nuñez

Cooper

Sanchez‐Moran

2021

Plants

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DNA topoisomerase II (TOPII) plays a very important role in DNA topology and in different biological processes such as DNA replication, transcription, repair, and chromosome condensation in higher eukaryotes. TOPII has been found to interact directly with a protein called topoisomerase II binding protein 1 (TopBP1) which also seems to have important roles in DNA replication and repair. In this study, we conducted different experiments to assess the roles of TopBP1 in DNA repair, mitosis, and meiosis, exploring the relationship between TOPII activity and TopBP1. We found that topbp1 mutant seedlings of Arabidopsis thaliana were hypersensitive to cisplatin treatment and the inhibition of TOPII with etoposide produced similar hypersensitivity levels. Furthermore, we recognised that there were no significant differences between the WT and topbp1 seedlings treated with cisplatin and etoposide together, suggesting that the hypersensitivity to cisplatin in the topbp1 mutant could be related to the functional interaction between TOPII and TopBP1. Somatic and meiotic anaphase bridges appeared in the topbp1 mutant at similar frequencies to those when TOPII was inhibited with merbarone, etoposide, or ICFR-187. The effects on meiosis of TOPII inhibition were produced at S phase/G2 stage, suggesting that catenanes could be produced at the onset of meiosis. Thus, if the processing of the catenanes is impaired, some anaphase bridges can be formed. Also, the appearance of anaphase bridges at first and second division is discussed.

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Section: Analysis Of Wt Male Meiosis Treated With Topii Inhibitorsmentioning

confidence: 99%

Section: Male Meiocytes Treated With Topii Inhibitors Show Anaphase B...mentioning

confidence: 99%

Section: Treatments With Topii Inhibitorsmentioning

confidence: 99%

See 1 more Smart Citation

The Role of DNA Topoisomerase Binding Protein 1 (TopBP1) in Genome Stability in Arabidopsis

Parra-Nuñez

Cooper

Sanchez‐Moran

2021

Plants

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“…Live-cell imaging techniques have been recently applied to A. thaliana meiosis but, until now, have mainly been used to study meiotic chromosome segregation 32 – 34 . Here, we applied the approach described by ref.…”

Section: Resultsmentioning

confidence: 99%

“…There is no precise description in plants of the RPMs nor of any machinery components involved in chromosome movement. We took the opportunity of recent advances in live imaging of meiosis in A. thaliana 32 – 34 to develop an approach to analyze meiotic prophase centromere dynamics in this model plant. Using 3D fluorescence time-lapse microscopy on male reproductive organs (anthers) and quantitative image analyses, we describe for the first time A. thaliana RPMs.…”

Section: Introductionmentioning

confidence: 99%

Rapid meiotic prophase chromosome movements in Arabidopsis thaliana are linked to essential reorganization at the nuclear envelope

Cromer,

Tiscareno-Andrade,

Lefranc

et al. 2024

Nat Commun

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Meiotic rapid prophase chromosome movements (RPMs) require connections between the chromosomes and the cytoskeleton, involving SUN (Sad1/UNC-84)-domain-containing proteins at the inner nuclear envelope (NE). RPMs remain significantly understudied in plants, with respect to their importance in the regulation of meiosis. Here, we demonstrate that Arabidopsis thaliana meiotic centromeres undergo rapid (up to 500 nm/s) and uncoordinated movements during the zygotene and pachytene stages. These centromere movements are not affected by altered chromosome organization and recombination but are abolished in the double mutant sun1 sun2. We also document the changes in chromosome dynamics and nucleus organization during the transition from leptotene to zygotene, including telomere attachment to SUN-enriched NE domains, bouquet formation, and nucleolus displacement, all of which were defective in sun1 sun2. These results establish A. thaliana as a model species for studying the functional implications of meiotic RPMs and demonstrate the mechanistic conservation of telomere-led RPMs in plants.

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Caught in the Act: Live-Cell Imaging of Plant Meiosis

et al. 2021

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Live-cell imaging is a powerful method to obtain insights into cellular processes, particularly with respect to their dynamics. This is especially true for meiosis, where chromosomes and other cellular components such as the cytoskeleton follow an elaborate choreography over a relatively short period of time. Making these dynamics visible expands understanding of the regulation of meiosis and its underlying molecular forces. However, the analysis of meiosis by live-cell imaging is challenging; specifically in plants, a temporally resolved understanding of chromosome segregation and recombination events is lacking. Recent advances in live-cell imaging now allow the analysis of meiotic events in plants in real time. These new microscopy methods rely on the generation of reporter lines for meiotic regulators and on the establishment of ex vivo culture and imaging conditions, which stabilize the specimen and keep it alive for several hours or even days. In this review, we combine an overview of the technical aspects of live-cell imaging in plants with a summary of outstanding questions that can now be addressed to promote live-cell imaging in Arabidopsis and other plant species and stimulate ideas on the topics that can be addressed in the context of plant meiotic recombination.

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

Cited by 3 publications

References 29 publications

The Role of DNA Topoisomerase Binding Protein 1 (TopBP1) in Genome Stability in Arabidopsis

The Role of DNA Topoisomerase Binding Protein 1 (TopBP1) in Genome Stability in Arabidopsis

Rapid meiotic prophase chromosome movements in Arabidopsis thaliana are linked to essential reorganization at the nuclear envelope

Caught in the Act: Live-Cell Imaging of Plant Meiosis

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