, Volume 128, Issue 3, pp 267–277 | Cite as

Spindle assembly without spindle pole body insertion into the nuclear envelope in fission yeast meiosis

  • Alberto Pineda-Santaella
  • Alfonso Fernández-ÁlvarezEmail author
Original Article


Centrosomes represent the major microtubule organizing center (MTOC) in eukaryotic cells and are responsible for nucleation of the spindle, the vehicle of chromosome segregation. In human female meiosis, however, spindle assembly occurs in the absence of centrosomes or other MTOCs and microtubules are nucleated around chromosomes. In yeast, spindle formation in mitosis and meiosis depends on the activity of spindle pole bodies (SPBs), the functional equivalents of centrosomes; thus, SPBs and centrosomes use similar machineries to assemble spindles. Here, we develop a system to explore the molecular mechanisms supporting acentrosomal spindle formation using fission yeast meiosis as a model scenario. We achieve this situation by removing access of the SPBs to the nucleus after their duplication. Under these conditions, we observe self-assembly-based spindle formation in the nuclear environment, conferring an ability to segregate chromosomes independently of the SPBs. Our results open the possibility to utilize the experimental advantages of fission yeast for insights into the molecular basis of acentrosomal spindle formation in meiosis.


Spindle Meiosis Fission yeast Centrosome Spindle pole body 



meiosis I


meiosis II


nuclear envelope


spindle pole body


nuclear envelope breakdown



We thank Julie Cooper, María Almuedo-Castillo, and Isabel Almudí for critical comments on the manuscript; Yesica García for technical support; and the CABD microscopy facility technician Katherina García for their helpful advice. We would like to thank the Genetics Department for their useful discussions and comments, especially Rafael R. Daga, Juan Jiménez, and Jose Ignacio Ibeas; and Víctor Manuel Carranco Fabre for the graphic design in the figures and technical support.

Funding information

This work was supported by Spanish Government (Ramon y Cajal program, RyC-2016-19659) to AF-A; AP-S by Pablo de Olavide University Fellowship (PPI1807). The CABD is an institution funded by Pablo de Olavide University, Consejo Superior de Investigaciones Científicas (CSIC) and Junta de Andalucía.

Supplementary material

412_2019_710_MOESM1_ESM.xlsx (11 kb)
ESM 1 (XLSX 11 kb)


  1. Bestul AJ, Yu Z, Unruh JR, Jaspersen SL (2017) Molecular model of fission yeast centrosome assembly determined by superresolution imaging. J Cell Biol 216:2409–2424PubMedPubMedCentralGoogle Scholar
  2. Blower MD, Nachury M, Heald R, Weis K (2005) A Rae1-containing ribonucleoprotein complex is required for mitotic spindle assembly. Cell 121:223–234PubMedGoogle Scholar
  3. Carazo-Salas RE, Guarguaglini G, Gruss OJ, Segref A, Karsenti E, Mattaj IW (1999) Generation of GTP-bound ran by RCC1 is required for chromatin-induced mitotic spindle formation. Nature 400:178–181PubMedGoogle Scholar
  4. Chikashige Y, Tsutsumi C, Yamane M, Okamasa K, Haraguchi T, Hiraoka Y (2006) Meiotic proteins bqt1 and bqt2 tether telomeres to form the bouquet arrangement of chromosomes. Cell 125:59–69PubMedGoogle Scholar
  5. Clarke PR, Zhang C (2008) Spatial and temporal coordination of mitosis by ran GTPase. Nat Rev Mol Cell Biol 9:464–477PubMedGoogle Scholar
  6. Courtois A, Schuh M, Ellenberg J, Hiiragi T (2012) The transition from meiotic to mitotic spindle assembly is gradual during early mammalian development. J Cell Biol 198:357–370PubMedPubMedCentralGoogle Scholar
  7. Ding R, West RR, Morphew DM, Oakley BR, McIntosh JR (1997) The spindle pole body of Schizosaccharomyces pombe enters and leaves the nuclear envelope as the cell cycle proceeds. Mol Biol Cell 8:1461–1479PubMedPubMedCentralGoogle Scholar
  8. Fennell A, Fernandez-Alvarez A, Tomita K, Cooper JP (2015) Telomeres and centromeres have interchangeable roles in promoting meiotic spindle formation. J Cell Biol 208:415–428PubMedPubMedCentralGoogle Scholar
  9. Fernandez-Alvarez A, Bez C, O'Toole ET, Morphew M, Cooper JP (2016) Mitotic nuclear envelope breakdown and spindle nucleation are controlled by interphase contacts between centromeres and the nuclear envelope. Dev Cell 39:544–559PubMedPubMedCentralGoogle Scholar
  10. Fernandez-Alvarez A, Cooper JP (2017) Chromosomes orchestrate their own liberation: nuclear envelope disassembly. Trends Cell Biol 27:255–265PubMedGoogle Scholar
  11. Flor-Parra I, Iglesias-Romero AB, Salas-Pino S, Lucena R, Jimenez J, Daga RR (2018) Importin alpha and vNEBD control meiotic spindle disassembly in fission yeast. Cell Rep 23:933–941PubMedGoogle Scholar
  12. Hagan I, Yanagida M (1995) The product of the spindle formation gene sad1+ associates with the fission yeast spindle pole body and is essential for viability. J Cell Biol 129:1033–1047PubMedGoogle Scholar
  13. Halpin D, Kalab P, Wang J, Weis K, Heald R (2011) Mitotic spindle assembly around RCC1-coated beads in Xenopus egg extracts. PLoS Biol 9:e1001225PubMedPubMedCentralGoogle Scholar
  14. Hashida-Okado T, Yasumoto R, Endo M, Takesako K, Kato I (1998) Isolation and characterization of the aureobasidin A-resistant gene, aur1R, on Schizosaccharomyces pombe: roles of Aur1p+ in cell morphogenesis. Curr Genet 33:38–45PubMedGoogle Scholar
  15. Hassold T, Hunt P (2001) To err (meiotically) is human: the genesis of human aneuploidy. Nat Rev Genet 2:280–291PubMedGoogle Scholar
  16. Heald R, Tournebize R, Blank T, Sandaltzopoulos R, Becker P, Hyman A, Karsenti E (1996) Self-organization of microtubules into bipolar spindles around artificial chromosomes in Xenopus egg extracts. Nature 382:420–425PubMedGoogle Scholar
  17. Hertig AT, Adams EC (1967) Studies on the human oocyte and its follicle. I. Ultrastructural and histochemical observations on the primordial follicle stage. J Cell Biol 34:647–675PubMedPubMedCentralGoogle Scholar
  18. Holubcova Z, Blayney M, Elder K, Schuh M (2015) Human oocytes. Error-prone chromosome-mediated spindle assembly favors chromosome segregation defects in human oocytes. Science 348:1143–1147PubMedPubMedCentralGoogle Scholar
  19. Kitamura E, Tanaka K, Komoto S, Kitamura Y, Antony C, Tanaka TU (2010) Kinetochores generate microtubules with distal plus ends: their roles and limited lifetime in mitosis. Dev Cell 18:248–259PubMedPubMedCentralGoogle Scholar
  20. Kume K, Kaneko S, Nishikawa K, Mizunuma M, Hirata D (2018) Role of nucleocytoplasmic transport in interphase microtubule organization in fission yeast. Biochem Biophys Res Commun 503:1160–1167PubMedGoogle Scholar
  21. Makarova M, Oliferenko S (2016) Mixing and matching nuclear envelope remodeling and spindle assembly strategies in the evolution of mitosis. Curr Opin Cell Biol 41:43–50PubMedGoogle Scholar
  22. Manandhar G, Schatten H, Sutovsky P (2005) Centrosome reduction during gametogenesis and its significance. Biol Reprod 72:2–13PubMedGoogle Scholar
  23. McGill M, Brinkley BR (1975) Human chromosomes and centrioles as nucleating sites for the in vitro assembly of microtubules from bovine brain tubulin. J Cell Biol 67:189–199PubMedGoogle Scholar
  24. Mikeladze-Dvali T, von Tobel L, Strnad P, Knott G, Leonhardt H, Schermelleh L, Gonczy P (2012) Analysis of centriole elimination during C. elegans oogenesis. Development 139:1670–1679PubMedPubMedCentralGoogle Scholar
  25. Mogessie B, Scheffler K, Schuh M (2018) Assembly and positioning of the oocyte meiotic spindle. Annu Rev Cell Dev Biol 34:381–403PubMedGoogle Scholar
  26. Mogessie B, Schuh M (2017) Actin protects mammalian eggs against chromosome segregation errors. Science 357:eaal1647PubMedGoogle Scholar
  27. Moreno S, Klar A, Nurse P (1991) Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol 194:795–823PubMedGoogle Scholar
  28. Nakamura Y, Arai A, Takebe Y, Masuda M (2011) A chemical compound for controlled expression of nmt1-driven gene in the fission yeast Schizosaccharomyces pombe. Anal Biochem 412:159–164PubMedGoogle Scholar
  29. Nam HJ, Naylor RM, van Deursen JM (2015) Centrosome dynamics as a source of chromosomal instability. Trends Cell Biol 25:65–73PubMedGoogle Scholar
  30. Pavin N, Tolic IM (2016) Self-organization and forces in the mitotic spindle. Annu Rev Biophys 45:279–298PubMedGoogle Scholar
  31. Pimenta-Marques A, Bento I, Lopes CA, Duarte P, Jana SC, Bettencourt-Dias M (2016) A mechanism for the elimination of the female gamete centrosome in Drosophila melanogaster. Science 353:aaf4866PubMedGoogle Scholar
  32. Rodrigues-Martins A, Riparbelli M, Callaini G, Glover DM, Bettencourt-Dias M (2008) From centriole biogenesis to cellular function: centrioles are essential for cell division at critical developmental stages. Cell Cycle 7:11–16PubMedGoogle Scholar
  33. Severson AF, von Dassow G, Bowerman B (2016) Oocyte meiotic spindle assembly and function. Curr Top Dev Biol 116:65–98PubMedGoogle Scholar
  34. Szollosi D, Calarco P, Donahue RP (1972) Absence of centrioles in the first and second meiotic spindles of mouse oocytes. J Cell Sci 11:521–541PubMedGoogle Scholar
  35. Tallada VA, Tanaka K, Yanagida M, Hagan IM (2009) The S. pombe mitotic regulator Cut12 promotes spindle pole body activation and integration into the nuclear envelope. J Cell Biol 185:875–888PubMedPubMedCentralGoogle Scholar
  36. Tamm T, Grallert A, Grossman EP, Alvarez-Tabares I, Stevens FE, Hagan IM (2011) Brr6 drives the Schizosaccharomyces pombe spindle pole body nuclear envelope insertion/extrusion cycle. J Cell Biol 195:467–484PubMedPubMedCentralGoogle Scholar
  37. Tomita K, Cooper JP (2007) The telomere bouquet controls the meiotic spindle. Cell 130:113–126PubMedGoogle Scholar
  38. Tournier F, Karsenti E, Bornens M (1989) Parthenogenesis in Xenopus eggs injected with centrosomes from synchronized human lymphoid cells. Dev Biol 136:321–329PubMedGoogle Scholar
  39. Vertii A, Hehnly H, Doxsey S (2016) The centrosome, a multitalented renaissance organelle. Cold Spring Harb Perspect Biol 8PubMedPubMedCentralGoogle Scholar
  40. Walczak CE, Vernos I, Mitchison TJ, Karsenti E, Heald R (1998) A model for the proposed roles of different microtubule-based motor proteins in establishing spindle bipolarity. Curr Biol 8:903–913PubMedGoogle Scholar
  41. West RR, Vaisberg EV, Ding R, Nurse P, McIntosh JR (1998) cut11(+): a gene required for cell cycle-dependent spindle pole body anchoring in the nuclear envelope and bipolar spindle formation in Schizosaccharomyces pombe. Mol Biol Cell 9:2839–2855PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Andalusian Center for Developmental Biology (CABD)Pablo de Olavide University/CSIC/Junta de AndalucíaSevilleSpain
  2. 2.Department of Molecular Biology and Biochemical EngineerPablo de Olavide UniversitySevilleSpain

Personalised recommendations