PMID- 28930998
OWN - NLM
STAT- MEDLINE
DCOM- 20180516
LR  - 20190911
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 127
DP  - 2017 Sep 11
TI  - In Vitro Ovule Cultivation for Live-cell Imaging of Zygote Polarization and 
      Embryo Patterning in Arabidopsis thaliana.
LID - 10.3791/55975 [doi]
LID - 55975
AB  - In most flowering plants, the zygote and embryo are hidden deep in the mother 
      tissue, and thus it has long been a mystery of how they develop dynamically; for 
      example, how the zygote polarizes to establish the body axis and how the embryo 
      specifies various cell fates during organ formation. This manuscript describes an 
      in vitro ovule culture method to perform live-cell imaging of developing zygotes 
      and embryos of Arabidopsis thaliana. The optimized cultivation medium allows 
      zygotes or early embryos to grow into fertile plants. By combining it with a 
      poly(dimethylsiloxane) (PDMS) micropillar array device, the ovule is held in the 
      liquid medium in the same position. This fixation is crucial to observe the same 
      ovule under a microscope for several days from the zygotic division to the late 
      embryo stage. The resulting live-cell imaging can be used to monitor the 
      real-time dynamics of zygote polarization, such as nuclear migration and 
      cytoskeleton rearrangement, and also the cell division timing and cell fate 
      specification during embryo patterning. Furthermore, this ovule cultivation 
      system can be combined with inhibitor treatments to analyze the effects of 
      various factors on embryo development, and with optical manipulations such as 
      laser disruption to examine the role of cell-cell communication.
FAU - Kurihara, Daisuke
AU  - Kurihara D
AD  - Division of Biological Science, Graduate School of Science, Nagoya University; 
      Higashiyama Live-Holonics Project, JST-ERATO, Nagoya University; 
      kuri@bio.nagoya-u.ac.jp.
FAU - Kimata, Yusuke
AU  - Kimata Y
AD  - Division of Biological Science, Graduate School of Science, Nagoya University.
FAU - Higashiyama, Tetsuya
AU  - Higashiyama T
AD  - Division of Biological Science, Graduate School of Science, Nagoya University; 
      Higashiyama Live-Holonics Project, JST-ERATO, Nagoya University; Institute of 
      Transformative Bio-Molecules (ITbM), Nagoya University.
FAU - Ueda, Minako
AU  - Ueda M
AD  - Division of Biological Science, Graduate School of Science, Nagoya University; 
      Institute of Transformative Bio-Molecules (ITbM), Nagoya University; 
      m-ueda@itbm.nagoya-u.ac.jp.
LA  - eng
PT  - Journal Article
PT  - Video-Audio Media
DEP - 20170911
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
SB  - IM
MH  - Arabidopsis/*embryology
MH  - Cell Differentiation/physiology
MH  - Microscopy, Polarization/*methods
MH  - Ovule/*growth & development
MH  - Zygote/physiology
PMC - PMC5752210
EDAT- 2017/09/21 06:00
MHDA- 2018/05/17 06:00
PMCR- 2019/09/11
CRDT- 2017/09/21 06:00
PHST- 2017/09/21 06:00 [entrez]
PHST- 2017/09/21 06:00 [pubmed]
PHST- 2018/05/17 06:00 [medline]
PHST- 2019/09/11 00:00 [pmc-release]
AID - 55975 [pii]
AID - 10.3791/55975 [doi]
PST - epublish
SO  - J Vis Exp. 2017 Sep 11;(127):55975. doi: 10.3791/55975.