Two-photon microscopic observation of cell-production dynamics in the developing mammalian neocortex in utero

Ryotaro Kawasoe, Tomoyasu Shinoda, Yuki Hattori, Mami Nakagawa, Trung Quang Pham, Yoshihiro Tanaka, Ken Sagou, Kanako Saito, Satoru Katsuki, Tomomi Kotani, Akihito Sano, Toshihiko Fujimori, Takaki Miyata

Research output: Contribution to journalArticlepeer-review

Abstract

Morphogenesis and organ development should be understood based on a thorough description of cellular dynamics. Recent studies have explored the dynamic behaviors of mammalian neural progenitor cells (NPCs) using slice cultures in which three-dimensional systems conserve in vivo-like environments to a considerable degree. However, live observation of NPCs existing truly in vivo, as has long been performed for zebrafish NPCs, has yet to be established in mammals. Here, we performed intravital two-photon microscopic observation of NPCs in the developing cerebral cortex of H2B-EGFP or Fucci transgenic mice in utero. Fetuses in the uterine sac were immobilized using several devices and were observed through a window made in the uterine wall and the amniotic membrane while monitoring blood circulation. Clear visibility was obtained to the level of 300 μm from the scalp surface of the fetus, which enabled us to quantitatively assess NPC behaviors, such as division and interkinetic nuclear migration, within a neuroepithelial structure called the ventricular zone at embryonic day (E) 13 and E14. In fetuses undergoing healthy monitoring in utero for 60 min, the frequency of mitoses observed at the apical surface was similar to those observed in slice cultures and in freshly fixed in vivo specimens. Although the rate and duration of successful in utero observations are still limited (33% for ≥10 min and 14% for 60 min), further improvements based on this study will facilitate future understanding of how organogenetic cellular behaviors occur or are pathologically influenced by the systemic maternal condition and/or maternal-fetal relationships.

Original languageEnglish
Pages (from-to)118-128
Number of pages11
JournalDevelopment Growth and Differentiation
Volume62
Issue number2
DOIs
Publication statusPublished - 01-02-2020
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Developmental Biology
  • Cell Biology

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