The Spindle Assembly Checkpoint Functions during Early Development in Non-Chordate Embryos
Abstract
1. Introduction
2. Materials and Methods
2.1. Gamete Collection and Fertilization
2.2. Drug Treatments
2.3. Generation of Mad1 and Mad2 Antibodies
2.4. Western Blot Analysis
2.5. Immunofluorescence
2.6. EdU (5-Ethynyl-2′-Deoxyuridine) Staining
2.7. Chromosome Spreads
2.8. Microinjection
2.9. Time-Lapse Microscopy
2.10. CellMask Staining
3. Results
3.1. Multispecies Survey Identifies Two Classes of Embryos with Different Mitotic Responses to Spindle Defects
3.2. The Mitotic Delay Observed in Jellyfish, Sea Urchin, and Mussel Embryos Depends on the SAC Kinase Mps1
3.3. Chordate Embryos Do Not Arrest in Mitosis in the Presence of Spindle Perturbations
3.4. SAC Competence Does Not Correlate with Cell Size across Species
3.5. Mad1, Mad2 and Mps1 Do Not Localize to Unattached Kinetochores in P. mammillata Early Embryos
4. Discussion
4.1. SAC Activity in Embryos Defines Two Classes of Animals
4.2. SAC Activity in Relation to Kinetochore and Cytoplasm Content
4.3. SAC Deficient Embryos as an Evolutionary Novelty in the Chordate Lineage
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Species | Oocyte Diameter (µm) | Chromosome Number (2n) | Kin/V2cell × 10−4 (kin/µm3) | Vnucleus (µm3) | Spindle Length (µm) | SAC Competence Reference | |
---|---|---|---|---|---|---|---|
Chordates | Xenopus laevis | 1000 | 36 | 0.0014 | 3053 [33] | 53.5 [34] | − [5] |
Danio rerio | 800 | 50 | 0.0037 | − [6] | |||
Ciona intestinalis | 140 | 28 | 0.39 | − this study | |||
Phallusia mammillata | 130 | 16 | 0.29 | 1345 | 20–30 | − this study | |
Branchiostoma lanceolatum | 130 | 38 [35] | 0.66 | − this study | |||
Echinoderms | Hacelia attenuata | 155 | 44 [36] | 0.45 | + this study | ||
Paracentrotus lividus | 90 | 36 [37] | 1.9 | 1562 | 18 [38] | + this study | |
Arbacia lixula | 80 | 44 [36] | 3.3 | + this study | |||
Strongylocentrotus purpuratus | 80 | 42 [39] | 3.1 | + this study | |||
Sphaerechinus granularis | 110 | 42 [36] | 1.2 | + this study | |||
Mollusks | Mytilus galloprovincialis | 65 | 28 [40] | Small = 6.6 Large = 2.5 | 2393 | + this study | |
Spisula solidissima | 60 | 28 | 6.7 | + [11] | |||
Arthropods | Drosophila melanogaster | 500 | 8 | 0.018 | + [41] | ||
Nematodes | Caenorhabditis elegans | 50 | 12 | AB = 22 [15] P1 = 17 [15] | 523 [42] | 13 [43] | + [44] |
Cnidarians | Clytia hemisphaerica | 210 | 30 | 0.12 | 30–35 | + this study |
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Chenevert, J.; Roca, M.; Besnardeau, L.; Ruggiero, A.; Nabi, D.; McDougall, A.; Copley, R.R.; Christians, E.; Castagnetti, S. The Spindle Assembly Checkpoint Functions during Early Development in Non-Chordate Embryos. Cells 2020, 9, 1087. https://doi.org/10.3390/cells9051087
Chenevert J, Roca M, Besnardeau L, Ruggiero A, Nabi D, McDougall A, Copley RR, Christians E, Castagnetti S. The Spindle Assembly Checkpoint Functions during Early Development in Non-Chordate Embryos. Cells. 2020; 9(5):1087. https://doi.org/10.3390/cells9051087
Chicago/Turabian StyleChenevert, Janet, Marianne Roca, Lydia Besnardeau, Antonella Ruggiero, Dalileh Nabi, Alex McDougall, Richard R. Copley, Elisabeth Christians, and Stefania Castagnetti. 2020. "The Spindle Assembly Checkpoint Functions during Early Development in Non-Chordate Embryos" Cells 9, no. 5: 1087. https://doi.org/10.3390/cells9051087
APA StyleChenevert, J., Roca, M., Besnardeau, L., Ruggiero, A., Nabi, D., McDougall, A., Copley, R. R., Christians, E., & Castagnetti, S. (2020). The Spindle Assembly Checkpoint Functions during Early Development in Non-Chordate Embryos. Cells, 9(5), 1087. https://doi.org/10.3390/cells9051087