Williams Jason S, Hsu Jessica Y, Rossi Christy Cortez, Artinger Kristin Bruk
Department of Craniofacial Biology, School of Dental Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA; Cell Biology, Stem Cells, and Development Graduate Program, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.
Department of Craniofacial Biology, School of Dental Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA; Pharmacology Graduate Program, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.
Dev Biol. 2018 Dec 1;444 Suppl 1(Suppl 1):S274-S286. doi: 10.1016/j.ydbio.2018.03.022. Epub 2018 Mar 29.
Melanocytes derive from neural crest cells, which are a highly migratory population of cells that play an important role in pigmentation of the skin and epidermal appendages. In most vertebrates, melanocyte precursor cells migrate solely along the dorsolateral pathway to populate the skin. However, zebrafish melanocyte precursors also migrate along the ventromedial pathway, in route to the yolk, where they interact with other neural crest derivative populations. Here, we demonstrate the requirement for zebrafish paralogs pcdh10a and pcdh10b in zebrafish melanocyte precursor migration. pcdh10a and pcdh10b are expressed in a subset of melanocyte precursor and somatic cells respectively, and knockdown and TALEN mediated gene disruption of pcdh10a results in aberrant migration of melanocyte precursors resulting in fully melanized melanocytes that differentiate precociously in the ventromedial pathway. Live cell imaging analysis demonstrates that loss of pchd10a results in a reduction of directed cell migration of melanocyte precursors, caused by both increased adhesion and a loss of cell-cell contact with other migratory neural crest cells. Also, we determined that the paralog pcdh10b is upregulated and can compensate for the genetic loss of pcdh10a. Disruption of pcdh10b alone by CRISPR mutagenesis results in somite defects, while the loss of both paralogs results in enhanced migratory melanocyte precursor phenotype and embryonic lethality. These results reveal a novel role for pcdh10a and pcdh10b in zebrafish melanocyte precursor migration and suggest that pcdh10 paralogs potentially interact for proper transient migration along the ventromedial pathway.
黑素细胞起源于神经嵴细胞，神经嵴细胞是一类具有高度迁移能力的细胞群体，在皮肤和表皮附属器的色素沉着中发挥重要作用。在大多数脊椎动物中，黑素细胞前体细胞仅沿着背外侧途径迁移以定居于皮肤。然而，斑马鱼的黑素细胞前体细胞也沿着腹内侧途径迁移，途中会到达卵黄，在那里它们与其他神经嵴衍生细胞群体相互作用。在此，我们证明了斑马鱼同源基因pcdh10a和pcdh10b在斑马鱼黑素细胞前体细胞迁移中的作用。pcdh10a和pcdh10b分别在一部分黑素细胞前体和体细胞中表达，敲低pcdh10a以及利用TALEN介导的基因破坏会导致黑素细胞前体细胞迁移异常，从而产生完全黑素化且在腹内侧途径中早熟分化的黑素细胞。活细胞成像分析表明，pchd10a的缺失导致黑素细胞前体细胞定向细胞迁移减少，这是由细胞黏附增加以及与其他迁移性神经嵴细胞的细胞间接触丧失所致。此外，我们确定同源基因pcdh10b上调并可补偿pcdh10a的基因缺失。单独通过CRISPR诱变破坏pcdh10b会导致体节缺陷，而两个同源基因均缺失则会导致迁移性黑素细胞前体的表型增强以及胚胎致死。这些结果揭示了pcdh10a和pcdh10b在斑马鱼黑素细胞前体细胞迁移中的新作用，并表明pcdh10同源基因可能相互作用以实现沿腹内侧途径的正常短暂迁移。
