Live imaging of C. elegans oocytes and early embryos

Laband, Kimberley; Lacroix, Benjamin; Edwards, Frances A.; Canman, Julie C.; Dumont, Julien

doi:10.1016/bs.mcb.2018.03.025

Cited by 28 publications

(31 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A detailed protocol for live imaging of C. elegans oocytes was used with minor modifications (Laband et al, 2018). Fertilised oocytes were dissected and mounted in 5 µl of L-15 blastomere culture medium (0.5 mg/ml Inulin; 25 mM HEPES, pH 7.5 in 60% Leibowitz L-15 medium and 20% heat-inactivated FBS) on 24×40 mm coverslips.…”

Section: Methodsmentioning

confidence: 99%

Sumoylation regulates protein dynamics during meiotic chromosome segregation in C. elegans oocytes

Pelisch

Borja

Jaffray

et al. 2019

Journal of Cell Science

View full text Add to dashboard Cite

Oocyte meiotic spindles in most species lack centrosomes and the mechanisms that underlie faithful chromosome segregation in acentrosomal meiotic spindles are not well understood. In C. elegans oocytes, spindle microtubules exert a poleward force on chromosomes that is dependent on the microtubule-stabilising protein CLS-2, the orthologue of the mammalian CLASP proteins. The checkpoint kinase BUB-1 and CLS-2 localise in the central spindle and display a dynamic localisation pattern throughout anaphase, but the signals regulating their anaphase-specific localisation remains unknown. We have shown previously that SUMO regulates BUB-1 localisation during metaphase I. Here, we found that SUMO modification of BUB-1 is regulated by the SUMO E3 ligase GEI-17 and the SUMO protease ULP-1. SUMO and GEI-17 are required for BUB-1 localisation between segregating chromosomes during early anaphase I. We also show that CLS-2 is subject to SUMO-mediated regulation; CLS-2 precociously localises in the midbivalent when either SUMO or GEI-17 are depleted. Overall, we provide evidence for a novel, SUMO-mediated control of protein dynamics during early anaphase I in oocytes.

show abstract

Section: Methodsmentioning

confidence: 99%

Sumoylation regulates protein dynamics during meiotic chromosome segregation in C. elegans oocytes

Pelisch

Borja

Jaffray

et al. 2019

Journal of Cell Science

View full text Add to dashboard Cite

show abstract

“…Worms were dissected to free embryos, which were then mounted and imaged as described in (Laband et al, 2018). Acquisitions were made on a spinning disk confocal microscope (Roper Scientific), using a CFI APO LBDA S × 60/NA1.4 oil objective and a CoolSNAP HQ2 CCD camera (Photometrics Scientific).…”

Section: Methodsmentioning

confidence: 99%

BUB-1 promotes amphitelic chromosome biorientation via multiple activities at the kinetochore

Edwards

Maton

Gareil

et al. 2018

eLife

Self Cite

View full text Add to dashboard Cite

Accurate chromosome segregation relies on bioriented amphitelic attachments of chromosomes to microtubules of the mitotic spindle, in which sister chromatids are connected to opposite spindle poles. BUB-1 is a protein of the Spindle Assembly Checkpoint (SAC) that coordinates chromosome attachment with anaphase onset. BUB-1 is also required for accurate sister chromatid segregation independently of its SAC function, but the underlying mechanism remains unclear. Here we show that, in Caenorhabditis elegans embryos, BUB-1 accelerates the establishment of non-merotelic end-on kinetochore-microtubule attachments by recruiting the RZZ complex and its downstream partner dynein-dynactin at the kinetochore. In parallel, BUB-1 limits attachment maturation by the SKA complex. This activity opposes kinetochore-microtubule attachment stabilisation promoted by CLS-2CLASP-dependent kinetochore-microtubule assembly. BUB-1 is therefore a SAC component that coordinates the function of multiple downstream kinetochore-associated proteins to ensure accurate chromosome segregation.

show abstract

“…A detailed protocol for live imaging of C. elegans oocytes was used with minor modifications (Laband et al, 2018). Fertilized oocytes were dissected and mounted in 5 µl of L-15 blastomere culture medium (0.5 mg/mL Inulin; 25 mM HEPES, pH 7.5 in 60% Leibowitz L-15 medium and 20% heat-Inactivated FBS) on 24×40 mm #1.5 coverslips.…”

Section: Live Imaging Of Oocytesmentioning

confidence: 99%

BUB-1 targets PP2A:B56 to regulate chromosome congression during meiosis I in C. elegans oocytes

Borja

Soubigou

Taylor

et al. 2020

eLife

View full text Add to dashboard Cite

Protein Phosphatase 2A (PP2A) is a heterotrimer composed of scaffolding (A), catalytic (C), and regulatory (B) subunits. PP2A complexes with B56 subunits are targeted by Shugoshin and BUBR1 to protect centromeric cohesion and stabilise kinetochore–microtubule attachments in yeast and mouse meiosis. In Caenorhabditis elegans, the closest BUBR1 orthologue lacks the B56-interaction domain and Shugoshin is not required for meiotic segregation. Therefore, the role of PP2A in C. elegans female meiosis is unknown. We report that PP2A is essential for meiotic spindle assembly and chromosome dynamics during C. elegans female meiosis. BUB-1 is the main chromosome-targeting factor for B56 subunits during prometaphase I. BUB-1 recruits PP2A:B56 to the chromosomes via a newly identified LxxIxE motif in a phosphorylation-dependent manner, and this recruitment is important for proper chromosome congression. Our results highlight a novel mechanism for B56 recruitment, essential for recruiting a pool of PP2A involved in chromosome congression during meiosis I.

show abstract

Live imaging of C. elegans oocytes and early embryos

Cited by 28 publications

References 30 publications

Sumoylation regulates protein dynamics during meiotic chromosome segregation in C. elegans oocytes

Sumoylation regulates protein dynamics during meiotic chromosome segregation in C. elegans oocytes

BUB-1 promotes amphitelic chromosome biorientation via multiple activities at the kinetochore

BUB-1 targets PP2A:B56 to regulate chromosome congression during meiosis I in C. elegans oocytes

Contact Info

Product

Resources

About