Long Term Ex Vivo Culture and Live Imaging of Drosophila Larval Imaginal Discs

被引:26
作者
Tsao, Chia-Kang [1 ,2 ]
Ku, Hui-Yu [1 ,2 ]
Lee, Yuan-Ming [1 ,2 ]
Huang, Yu-Fen [1 ,2 ,3 ]
Sun, Yi Henry [1 ,2 ]
机构
[1] Acad Sinica, Inst Mol Biol, Taipei, Taiwan
[2] Natl Yang Ming Univ, Inst Genom Sci, Taipei, Taiwan
[3] Howard Hughes Med Inst, Janelia Farm Res Campus, Ashburn, VA USA
来源
PLOS ONE | 2016年 / 11卷 / 09期
关键词
CELL DIVISIONS; MYOSIN-II; MIGRATION; DIFFERENTIATION; PROLIFERATION; LOCALIZATION; GROWTH; MORPHOGENESIS; BOUNDARY; PATTERN;
D O I
10.1371/journal.pone.0163744
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Continuous imaging of live tissues provides clear temporal sequence of biological events. The Drosophila imaginal discs have been popular experimental subjects for the study of a wide variety of biological phenomena, but long term culture that allows normal development has not been satisfactory. Here we report a culture method that can sustain normal development for 18 hours and allows live imaging. The method is validated in multiple discs and for cell proliferation, differentiation and migration. However, it does not support disc growth and cannot support cell proliferation for more than 7 to 12 hr. We monitored the cellular behavior of retinal basal glia in the developing eye disc and found that distinct glia type has distinct properties of proliferation and migration. The live imaging provided direct proof that wrapping glia differentiated from existing glia after migrating to the anterior front, and unexpectedly found that they undergo endoreplication before wrapping axons, and their nuclei migrate up and down along the axons. UV-induced specific labeling of a single carpet glia also showed that the two carpet glia membrane do not overlap and suggests a tiling or repulsion mechanism between the two cells. These findings demonstrated the usefulness of an ex vivo culture method and live imaging.
引用
收藏
页数:19
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