title: Visualization of craniofacial development in the sox10: kaede transgenic zebrafish line using time-lapse confocal microscopy.
abstract: Vertebrate palatogenesis is a highly choreographed and complex developmental process, which involves migration of cranial neural crest (CNC) cells, convergence and extension of facial prominences, and maturation of the craniofacial skeleton. To study the contribution of the cranial neural crest to specific regions of the zebrafish palate a sox10: kaede transgenic zebrafish line was generated. Sox10 provides lineage restriction of the kaede reporter protein to the neural crest, thereby making the cell labeling a more precise process than traditional dye or reporter mRNA injection. Kaede is a photo-convertible protein that turns from green to red after photo activation and makes it possible to follow cells precisely. The sox10: kaede transgenic line was used to perform lineage analysis to delineate CNC cell populations that give rise to maxillary versus mandibular elements and illustrate homology of facial prominences to amniotes. This protocol describes the steps to generate a live time-lapse video of a sox10: kaede zebrafish embryo. Development of the ethmoid plate will serve as a practical example. This protocol can be applied to making a time-lapse confocal recording of any kaede or similar photoconvertible reporter protein in transgenic zebrafish. Furthermore, it can be used to capture not only normal, but also abnormal development of craniofacial structures in the zebrafish mutants.
authors.affiliation: Center for Regenerative Medicine, Massachusetts General Hospital.
authors.forename: Lisa
authors.initials: L
authors.lastname: Gfrerer
authors.forename: Max
authors.initials: M
authors.lastname: Dougherty
authors.forename: Eric C
authors.initials: EC
authors.lastname: Liao
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D005004:Ethmoid Bone / Q000737:chemistry / Q000196:embryology*; D008164:Luminescent Proteins; D018613:Microscopy, Confocal / Q000379:methods*; D009432:Neural Crest / Q000737:chemistry / Q000166:cytology / Q000196:embryology; D055757:SOXE Transcription Factors / Q000235:genetics; D059008:Time-Lapse Imaging / Q000379:methods*; D015027:Zebrafish / Q000196:embryology*; D029961:Zebrafish Proteins / Q000235:genetics
chemical_list: D008164:Luminescent Proteins; D055757:SOXE Transcription Factors; D029961:Zebrafish Proteins; C442229:sox10 protein, zebrafish