Quantitative analysis of sensitivity to a Wnt3a gradient in determination of the pole-to-pole axis of mitotic cells by using a microfluidic device

Takumi Hiraiwa, Yuichiro Nakai, Takahiro G. Yamada, Ryuichi Tanimoto, Hiroshi Kimura, Yoshinori Matsumoto, Norihisa Miki, Noriko Hiroi, Akira Funahashi

Research output: Contribution to journalArticle

Abstract

Proper determination of the cell division axis is essential during development. Wnt3a is a known regulator of the cell division axis; however, the sensitivity of cells to Wnt3a signalling and its role in determining the cell division axis have not been measured to date. To address this gap, we took advantage of the asymmetric distribution of outer dense fibre 2 (ODF2/cenexin) proteins on centrosomes in dividing cells. To precisely quantify the sensitivity of cells to Wnt3a signalling, we developed a microfluidic cell culture device, which can produce a quantitative gradient of signalling molecules. We confirmed that mitotic SH-SY5Y neuroblastoma cells could detect a 2.5 ~ 5 × 10−3 nm·μm−1 Wnt3a concentration gradient and demonstrated that this gradient is sufficient to affect the determination of the pole-to-pole axis of cell division during the later stages of mitosis.

Original languageEnglish
Pages (from-to)1920-1935
Number of pages16
JournalFEBS Open Bio
Volume8
Issue number12
DOIs
Publication statusPublished - 2018 Dec

Keywords

  • Wnt3a
  • microfluidics
  • neuroblastoma
  • pole-to-pole axis
  • quantitative gradient ODF2

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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