Department of Genetics, Washington University, St. Louis, Missouri, United States of America.
PLoS One. 2013 Jul 9;8(7):e67801. doi: 10.1371/journal.pone.0067801. Print 2013.
In order to facilitate understanding of pigment cell biology, we developed a method to concomitantly purify melanocytes, iridophores, and retinal pigmented epithelium from zebrafish, and analyzed their transcriptomes. Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co-enrichment in melanocytes and retinal pigmented epithelium, as well as in melanocytes and iridophores. We found 214 genes co-enriched in melanocytes and retinal pigmented epithelium, indicating the shared functions of melanin-producing cells. We found 62 genes significantly co-enriched in melanocytes and iridophores, illustrative of their shared developmental origins from the neural crest. This is also the first analysis of the iridophore transcriptome. Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine-based reflective pigment. We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate-limiting substrate for purine synthesis, phosphoribosyl pyrophosphate, thus constituting a guanine cycle. The purification procedure and expression analysis described here, along with the accompanying transcriptome-wide expression data, provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types, and will be a useful resource for further studies of pigment cell biology.
为了便于理解色素细胞生物学,我们开发了一种从斑马鱼中同时纯化黑素细胞、虹彩细胞和视网膜色素上皮细胞的方法,并分析了它们的转录组。比较这些细胞类型和整个胚胎的表达数据,使我们能够揭示黑素细胞和视网膜色素上皮细胞以及黑素细胞和虹彩细胞中的基因表达共富集。我们发现 214 个基因在黑素细胞和视网膜色素上皮细胞中共同富集,表明产生黑色素的细胞具有共同的功能。我们发现 62 个基因在黑素细胞和虹彩细胞中显著共同富集,表明它们具有共同的神经嵴起源。这也是虹彩细胞转录组的首次分析。对虹彩细胞的基因表达分析显示,大量特异性酶的富集有助于协调其基于鸟嘌呤的反射性色素的产生。我们推测,几个代谢途径中特定酶的协调上调回收嘌呤合成的限速底物,5-磷酸核糖基焦磷酸,从而构成鸟嘌呤循环。本文描述的纯化程序和表达分析,以及伴随的全转录组表达数据,为多种纯化的斑马鱼色素细胞类型提供了第一个 mRNA 测序数据,将成为色素细胞生物学进一步研究的有用资源。