The Fugu tyrp1 promoter directs specific GFP expression in zebrafish: tools to study the RPE and the neural crest-derived melanophores.
In vertebrates, pigment cells account for a small percentage of the total cell population and they intermingle with other cell types. This makes it difficult to isolate them for analyzes of their functions in the context of development. To alleviate such difficulty, we generated two stable transgenic zebrafish lines (pt101 and pt102) that express green fluorescent protein (GFP) in melanophores under the control of the 1 kb Fugu tyrp1 promoter. In pt101, GFP is expressed in both retinal pigment epithelium (RPE) cells and the neural crest-derived melanophores (NCDM), whereas in pt102, GFP is predominately expressed in the NCDM. Our results indicate that the Fugu tyrp1 promoter can direct transgene expression in a cell-type-specific manner in zebrafish. In addition, our findings provide evidence supporting differential regulations of melanin-synthesizing genes in RPE cells and the NCDM in zebrafish. Utilizing the varying GFP expression levels in these fish, we have isolated melanophores via flow cytometry and revealed the capability of sorting the NCDM from RPE cells as well. Thus, these transgenic lines are useful tools to study melanophores in zebrafish.
Zou
Jian
J
Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.
Beermann
Friedrich
F
Wang
Jianxin
J
Kawakami
Koichi
K
Wei
Xiangyun
X
D000818:Animals; D030801:Animals, Genetically Modified; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D019070:Cell Lineage / Q000235:genetics; D004626:Embryology / Q000379:methods; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000235:genetics*; D008297:Male; D008547:Melanophores / Q000166:cytology / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods; D009432:Neural Crest / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics*; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D023741:Takifugu; D019076:Transgenes / Q000235:genetics; D015027:Zebrafish
D049452:Green Fluorescent Proteins; D010088:Oxidoreductases; C064056:tyrosinase-related protein-1