File:Mouse neural tube 01.mp4

Mouse Neural Tube Closure

Live imaging of the sealing midbrain–hindbrain neuropore (MHNP) by closures I and II. Time-lapse video of a SCAT3tg/+ embryo undergoing MHNP closure, as shown in Fig. 2 F. ECFP images are shown. MHNP was sealed by closures I and II. Live imaging was performed on an inverted confocal microscope (TCS SP5). Frames were taken at intervals of 4 min for 14 h.

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Reference

<pubmed>22162136 </pubmed>| PMC3241723 | J Cell Biol.

Copyright

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Video 2. http://jcb.rupress.org/content/suppl/2011/12/08/jcb.201104057.DC1/JCB_201104057_V2.mov

Live imaging of apoptosis in a novel transgenic mouse highlights its role in neural tube closure

J Cell Biol. 2011 Dec 12;195(6):1047-60. doi: 10.1083/jcb.201104057.

Yamaguchi Y, Shinotsuka N, Nonomura K, Takemoto K, Kuida K, Yosida H, Miura M. Source Department of Genetics, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan. bunbun@mol.f.u-tokyo.ac.jp

Abstract

Many cells die during development, tissue homeostasis, and disease. Dysregulation of apoptosis leads to cranial neural tube closure (NTC) defects like exencephaly, although the mechanism is unclear. Observing cells undergoing apoptosis in a living context could help elucidate their origin, behavior, and influence on surrounding tissues, but few tools are available for this purpose, especially in mammals. In this paper, we used insulator sequences to generate a transgenic mouse that stably expressed a genetically encoded fluorescence resonance energy transfer (FRET)-based fluorescent reporter for caspase activation and performed simultaneous time-lapse imaging of apoptosis and morphogenesis in living embryos. Live FRET imaging with a fast-scanning confocal microscope revealed that cells containing activated caspases showed typical and nontypical apoptotic behavior in a region-specific manner during NTC. Inhibiting caspase activation perturbed and delayed the smooth progression of cranial NTC, which might increase the risk of exencephaly. Our results suggest that caspase-mediated cell removal facilitates NTC completion within a limited developmental window.

PMID 22162136