Loss of CENPF leads to developmental failure in mouse embryos

Cell Cycle. 2019 Oct;18(20):2784-2799. doi: 10.1080/15384101.2019.1661173. Epub 2019 Sep 3.

Abstract

Aneuploidy caused by abnormal chromosome segregation during early embryo development leads to embryonic death or congenital malformation. Centromere protein F (CENPF) is a member of centromere protein family that regulates chromosome segregation during mitosis. However, its necessity in early embryo development has not been fully investigated. In this study, expression and function of CENPF was investigated in mouse early embryogenesis. Detection of CENPF expression and localization revealed a cytoplasm, spindle and nuclear membrane related dynamic pattern throughout mitotic progression. Farnesyltransferase inhibitor (FTI) was employed to inhibit CENPF farnesylation in zygotes. The results showed that CENPF degradation was inhibited and its specific localization on nuclear membranes in morula and blastocyst vanished after FTI treatment. Also, CAAX motif mutation leads to failure of CENPF-C630 localization in morula and blastocyst. These results indicate that farnesylation plays a key role during CENPF degradation and localization in early embryos. To further assess CENPF function in parthenogenetic or fertilized embryos development, morpholino (MO) and Trim-Away were used to disturb CENPF function. CENPF knockdown in Metaphase II (MII) oocytes, zygotes or embryos with MO approach resulted in failure to develop into morulae and blastocysts, revealing its indispensable role in both parthenogenetic and fertilized embryos. Disturbing of CENPF with Trim-Away approach in zygotes resulted in impaired development of 2-cell and 4-cell, but did not affect the morula and blastocyst formation because of the recovered expression of CENPF. Taken together, our data suggest CENPF plays an important role during early embryonic development in mice. Abbreviation: CENPF: centromere protein F; MO: morpholino; FTI: Farnesyltransferase inhibitor; CENPE: centromere protein E; IVF: in vitro fertilization; MII: metaphase II; SAC: spindle assembly checkpoint; Mad1: mitotic arrest deficient 1; BUB1: budding uninhibited by benzimidazole 1; BUBR1: BUB1 mitotic checkpoint serine/threonine kinase B; Cdc20: cell division cycle 20.

Keywords: CENPF; Morpholino; Trim-Away; early embryo development; farnesylation.

MeSH terms

  • Animals
  • Blastocyst / metabolism*
  • Centromere / metabolism
  • Chromosomal Proteins, Non-Histone / chemistry
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism*
  • Embryo, Mammalian / embryology
  • Embryo, Mammalian / metabolism*
  • Embryonic Development / genetics*
  • Farnesyltranstransferase / antagonists & inhibitors
  • Female
  • Gene Knockdown Techniques
  • Metaphase / genetics
  • Mice
  • Mice, Inbred ICR
  • Microfilament Proteins / chemistry
  • Microfilament Proteins / genetics
  • Microfilament Proteins / metabolism*
  • Morpholinos / genetics
  • Morpholinos / pharmacology
  • Morula / metabolism
  • Oocytes / growth & development
  • Oocytes / metabolism*
  • Parthenogenesis / genetics
  • Piperidines / pharmacology
  • Pregnancy
  • Prenylation
  • Pyridines / pharmacology
  • Zygote / growth & development
  • Zygote / metabolism*

Substances

  • Chromosomal Proteins, Non-Histone
  • Microfilament Proteins
  • Morpholinos
  • Piperidines
  • Pyridines
  • centromere protein F
  • Farnesyltranstransferase
  • lonafarnib

Grants and funding

This work was supported by the NSF of China (31871508, 31960158, 31671560 to C.G.L.; 31900596 to C.J.Z.), Major Projects of Natural Science Foundation of Inner Mongolia Autonomous Region (No. 2019ZD008 to C.G.L.), The Funding of Top Disciplines Development in Inner Mongolia University, Open Project of the State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock (30500-1220020402 to C.G.L.), Science and Technology Major Project of Inner Mongolia Autonomous Region of China to the State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock (30500-518390216 to C.J.Z.), Research Foundation for Advanced Talents of Inner Mongolia University (30500-5185142 to C.J.Z.).