Spatio-Temporal Tracking of Melanocytic Differentiation and Migration of Cranial Neural Crest in Zebrafish Embryonic Development
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We set out to characterize a population of midbrain fluorescing cells in zebrafish that appeared to be of melanophore lineage and in doing so discovered a potential role for sox10 expression in the zebrafish central nervous system glial cell network. A decrease in the signal of interest correlated with increased inhibition of melanin production by PTU (1-phenyl-2-thioreau) treatment; however, the autofluorescence signal at question was determined not to be associated with melanin biosynthesis after wild type time lapse imaging experiments showed the absence of said signal, and it was shown that only after fixation of the embryos was the signal produced. To characterize these strongly autofluorescing cells, melanophore gene expression was visualized using transgenic reporters for the transcriptions factors sox10 and mitfa, a neural crest marker and a melanocyte identity regulator, respectively, via broadband ultrashort pulse excitation centered at 800 nm with FWHM of 133 nm to determine the genetic identity of the autofluorescence source via colocalization of signals. Time lapse imaging was performed to collect 4-D data sets of fluorescence reporting of the neural crest marker sox10 and melanophore identity regulator mitfa within the zebrafish midbrain between the prim-5 and prim-25 stage, revealing a novel population of sox10 expressing cells within the neuroepithelium that proliferates and migrates within the ventral midbrain tissue towards the developing nasal cavity, suggesting a role sox10 may play in oligodendrocyte regulatory networks in this region. Gene expression visualization of mitfa in the same region displayed no neuroepithelial expression as expected.
two photon microscopy
ultrashort pulse microscopy
Robertson, Benjamin Patrick (2016). Spatio-Temporal Tracking of Melanocytic Differentiation and Migration of Cranial Neural Crest in Zebrafish Embryonic Development. Master's thesis, Texas A & M University. Available electronically from