Small molecule inhibitor of formin homology 2 domains (SMIFH2) reveals the roles of the formin family of proteins in spindle assembly and asymmetric division in mouse oocytes

PLoS One. 2015 Apr 2;10(4):e0123438. doi: 10.1371/journal.pone.0123438. eCollection 2015.

Abstract

Dynamic actin reorganization is the main driving force for spindle migration and asymmetric cell division in mammalian oocytes. It has been reported that various actin nucleators including Formin-2 are involved in the polarization of the spindle and in asymmetric cell division. In mammals, the formin family is comprised of 15 proteins. However, their individual roles in spindle migration and/or asymmetric division have not been elucidated yet. In this study, we employed a newly developed inhibitor for formin family proteins, small molecule inhibitor of formin homology 2 domains (SMIFH2), to assess the functions of the formin family in mouse oocyte maturation. Treatment with SMIFH2 during in vitro maturation of mouse oocytes inhibited maturation by decreasing cytoplasmic and cortical actin levels. In addition, treatment with SMIFH2, especially at higher concentrations (500 μM), impaired the proper formation of meiotic spindles, indicating that formins play a role in meiotic spindle formation. Knockdown of the mDia2 formins caused a similar decrease in oocyte maturation and abnormal spindle morphology, mimicking the phenotype of SMIFH2-treated cells. Collectively, these results suggested that besides Formin-2, the other proteins of the formin, including mDia family play a role in asymmetric division and meiotic spindle formation in mammalian oocytes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Animals
  • Asymmetric Cell Division / genetics*
  • Carrier Proteins / genetics*
  • Carrier Proteins / metabolism
  • Cells, Cultured
  • Formins
  • Meiosis
  • Mice
  • Microtubule-Associated Proteins / genetics*
  • Microtubule-Associated Proteins / metabolism
  • NADPH Dehydrogenase / genetics*
  • NADPH Dehydrogenase / metabolism
  • Oocytes / cytology
  • Oocytes / drug effects
  • Oocytes / growth & development*
  • RNA Interference
  • RNA, Small Interfering
  • Spindle Apparatus / metabolism*
  • Thiones / pharmacology*
  • Tropomyosin / metabolism
  • Uracil / analogs & derivatives*
  • Uracil / pharmacology

Substances

  • Actins
  • Carrier Proteins
  • Diap1 protein, mouse
  • Formins
  • Microtubule-Associated Proteins
  • RNA, Small Interfering
  • SMIFH2 compound
  • Thiones
  • Tropomyosin
  • Uracil
  • Dia2 protein, mouse
  • NADPH Dehydrogenase

Grants and funding

Supported by grants from the Next-Generation BioGreen 21 Program (PJ011206 and PJ011126, Rural Development Administration, Republic of Korea. http://www.rda.go.kr/foreign/eng/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.