Long Term Ex Vivo Culture and Live Imaging of Drosophila Larval Imaginal Discs.
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.
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan.|Institute of Genomic Sciences, National Yang-Ming University, Taipei, Taiwan.
Chia-Kang
CK
Tsao
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan.|Institute of Genomic Sciences, National Yang-Ming University, Taipei, Taiwan.
Hui-Yu
HY
Ku
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan.|Institute of Genomic Sciences, National Yang-Ming University, Taipei, Taiwan.
Yuan-Ming
YM
Lee
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan.|Institute of Genomic Sciences, National Yang-Ming University, Taipei, Taiwan.
Yu-Fen
YF
Huang
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan.|Institute of Genomic Sciences, National Yang-Ming University, Taipei, Taiwan.
Yi Henry
http://orcid.org/0000-0001-8279-5270
YH
Sun
Neuron. 1998 Jan;20(1):83-93
9459444
Development. 2014 Jun;141(11):2339-48
24866118
Mol Cell. 2003 Oct;12(4):1051-8
14580354
Genes Dev. 1998 Feb 1;12(3):435-46
9450936
Proc Natl Acad Sci U S A. 2010 Aug 10;107(32):14217-22
20660765
J Cell Biol. 2010 Mar 8;188(5):735-49
20194639
Wiley Interdiscip Rev Dev Biol. 2013 Jul;2(4):545-57
24014422
Methods Enzymol. 2012;506:407-13
22341235
PLoS One. 2012;7(1):e29869
22291895
J Neurosci. 2007 Nov 28;27(48):13130-9
18045907
Nature. 2009 Jan 22;457(7228):495-9
19029882
Curr Opin Genet Dev. 2007 Aug;17(4):309-13
17618111
Proc Natl Acad Sci U S A. 1996 Oct 15;93(21):11687-92
8876197
J Neurobiol. 1995 Nov;28(3):363-80
8568517
Dev Biol. 2003 Apr 15;256(2):389-402
12679111
J Vis Exp. 2009 Jul 09;(29):null
19590493
PLoS One. 2014 Sep 09;9(9):e107333
25203426
Development. 1999 Aug;126(15):3285-92
10393108
Nature. 2006 Aug 31;442(7106):1038-41
16900102
Genetics. 2007 Feb;175(2):567-84
17151243
Development. 2000 Mar;127(6):1151-9
10683169
Science. 1996 Oct 11;274(5285):252-5
8824191
J Embryol Exp Morphol. 1966 Jun;15(3):271-9
5964277
Neuron. 1994 Feb;12(2):423-31
8110466
Development. 1994 Sep;120(9):2609-18
7956836
Science. 1977 Apr 22;196(4288):438-40
403606
J Exp Zool. 1964 Jun;156:91-103
14189923
Development. 2013 Oct;140(19):4051-9
24046320
Mol Biol Cell. 2004 Feb;15(2):838-50
14657248
J Embryol Exp Morphol. 1970 Feb;23(1):53-69
5503855
Dev Cell. 2014 Apr 14;29(1):112-27
24684830
Dev Dyn. 2006 Nov;235(11):3051-8
17013876
Development. 2013 Feb 1;140(3):667-74
23293298
Nature. 1985 Jan 10-18;313(5998):138-41
3917556
Genes Dev. 2011 Jan 15;25(2):131-6
21245166
Exp Cell Res. 1982 Jun;139(2):297-307
6806111
Curr Biol. 2005 Sep 20;15(18):1640-4
16169485
Nat Commun. 2014 Jul 28;5:4484
25066644
Proc Natl Acad Sci U S A. 1988 Aug;85(15):5454-8
3399501
Cell Death Differ. 2010 Mar;17(3):534-9
19960024
Dev Biol. 1973 Aug;33(2):441-56
4207978
Curr Biol. 2005 May 24;15(10 ):955-62
15916953
Dev Biol. 2008 Jan 15;313(2):739-51
18082159
Science. 2012 May 11;336(6082):724-7
22499807
Cell Rep. 2014 Apr 24;7(2):588-98
24726363