To image adherens junction dynamics and track junction
clusters during apical constriction, the following format can be
tested first: physical size of each pixel: 0.12 μm; digital dimension of each image: 512 Â 256; Z stack depth: 7 μm with
0.5 μm interval; temporal interval: 5 s. Z Piezo driver is helpful
to image at such a high temporal resolution. Bidirectional
mode may be used depending on the speed of the microscope.
Acknowledgments
We would like to thank Rolin Sauceda for comments on the manuscript. The research in Weng lab is supported by UNLV startup and
the National Institute of Health (R00 HD088764).
References
1. Thiery JP, Sleeman JP (2006) Complex networks orchestrate epithelial-mesenchymal transitions. Nat Rev Mol Cell Biol 7:131–142
2. Lambert AW, Pattabiraman DR, Weinberg RA
(2017) Emerging biological principles of
metastasis. Cell 168:670–691
3. de Craene B, Berx G (2013) Regulatory networks defining EMT during cancer initiation
and progression. Nat Rev Cancer 13:97–110
4. Li W, Kang Y (2016) Probing the fifty shades of
EMT in metastasis. Trends Cancer 2:65–67
5. Dongre A, Weinberg RA (2019) New insights
into the mechanisms of epithelial–mesenchymal transition and implications for cancer. Nat
Rev Mol Cell Biol 20:69–84
6. Nieto MA, Huang RYYJ, Jackson RAA, Thiery
JPP (2016) Emt: 2016. Cell 166:21–45
7. Weng M, Wieschaus E (2016) Myosindependent remodeling of adherens junctions
protects junctions from snail-dependent disassembly. J Cell Biol 212:219–229
8. Weng M, Wieschaus E (2017) Polarity protein
Par3/bazooka follows myosin-dependent
junction repositioning. Dev Biol 422:125–134
9. Tepass U, Hartenstein V (1994) The development of cellular junctions in the drosophila
embryo. Dev Biol 161(2):563–596. https://
doi.org/10.1006/dbio.1994.1054
10. Lamouille S, Xu J, Derynck R (2014) Molecular mechanisms of epithelial-mesenchymal
transition. Nat Rev Mol Cell Biol 15:178–196
11. Bothma JP, Norstad MR, Alamos S, Garcia HG
(2018) LlamaTags: a versatile tool to image
transcription factor dynamics in live embryos.
Cell 173(7):1810–1822.e16. https://doi.org/
10.1016/j.cell.2018.03.069
12. Lim J, Thiery JP (2012) Epithelialmesenchymal transitions: insights from development. Development 139:3471–3486
13. Shishido E, Higashijima S, Emori Y, Saigo K
(1993) Two FGF-receptor homologues of drosophila: one is expressed in mesodermal primordium in early embryos. Development
117:751–761
14. Vincent S, Wilson R, Coelho C, Affolter M,
Leptin M (1998) The drosophila protein Dof
is specifically required for FGF signaling. Mol
Cell 2:515–525
15. Manning AJ, Peters KA, Peifer M, Rogers SL
(2013) Regulation of epithelial morphogenesis
by the gprotein-coupled receptor mist and its
ligand fog. Sci Signal. https://doi.org/10.
1126/scisignal.2004427
Live Imaging of EMT in Chick Neural Crest
77
clusters during apical constriction, the following format can be
tested first: physical size of each pixel: 0.12 μm; digital dimension of each image: 512 Â 256; Z stack depth: 7 μm with
0.5 μm interval; temporal interval: 5 s. Z Piezo driver is helpful
to image at such a high temporal resolution. Bidirectional
mode may be used depending on the speed of the microscope.
Acknowledgments
We would like to thank Rolin Sauceda for comments on the manuscript. The research in Weng lab is supported by UNLV startup and
the National Institute of Health (R00 HD088764).
References
1. Thiery JP, Sleeman JP (2006) Complex networks orchestrate epithelial-mesenchymal transitions. Nat Rev Mol Cell Biol 7:131–142
2. Lambert AW, Pattabiraman DR, Weinberg RA
(2017) Emerging biological principles of
metastasis. Cell 168:670–691
3. de Craene B, Berx G (2013) Regulatory networks defining EMT during cancer initiation
and progression. Nat Rev Cancer 13:97–110
4. Li W, Kang Y (2016) Probing the fifty shades of
EMT in metastasis. Trends Cancer 2:65–67
5. Dongre A, Weinberg RA (2019) New insights
into the mechanisms of epithelial–mesenchymal transition and implications for cancer. Nat
Rev Mol Cell Biol 20:69–84
6. Nieto MA, Huang RYYJ, Jackson RAA, Thiery
JPP (2016) Emt: 2016. Cell 166:21–45
7. Weng M, Wieschaus E (2016) Myosindependent remodeling of adherens junctions
protects junctions from snail-dependent disassembly. J Cell Biol 212:219–229
8. Weng M, Wieschaus E (2017) Polarity protein
Par3/bazooka follows myosin-dependent
junction repositioning. Dev Biol 422:125–134
9. Tepass U, Hartenstein V (1994) The development of cellular junctions in the drosophila
embryo. Dev Biol 161(2):563–596. https://
doi.org/10.1006/dbio.1994.1054
10. Lamouille S, Xu J, Derynck R (2014) Molecular mechanisms of epithelial-mesenchymal
transition. Nat Rev Mol Cell Biol 15:178–196
11. Bothma JP, Norstad MR, Alamos S, Garcia HG
(2018) LlamaTags: a versatile tool to image
transcription factor dynamics in live embryos.
Cell 173(7):1810–1822.e16. https://doi.org/
10.1016/j.cell.2018.03.069
12. Lim J, Thiery JP (2012) Epithelialmesenchymal transitions: insights from development. Development 139:3471–3486
13. Shishido E, Higashijima S, Emori Y, Saigo K
(1993) Two FGF-receptor homologues of drosophila: one is expressed in mesodermal primordium in early embryos. Development
117:751–761
14. Vincent S, Wilson R, Coelho C, Affolter M,
Leptin M (1998) The drosophila protein Dof
is specifically required for FGF signaling. Mol
Cell 2:515–525
15. Manning AJ, Peters KA, Peifer M, Rogers SL
(2013) Regulation of epithelial morphogenesis
by the gprotein-coupled receptor mist and its
ligand fog. Sci Signal. https://doi.org/10.
1126/scisignal.2004427
Live Imaging of EMT in Chick Neural Crest
77