Marine Biology Research Division, Scripps Institution of Oceanography, University of California, San Diego, California 92093-0202, USA.
Dev Dyn. 2012 Jun;241(6):1111-24. doi: 10.1002/dvdy.23786. Epub 2012 May 2.
ATP-binding cassette (ABC) transporters are membrane proteins that regulate intracellular concentrations of myriad compounds and ions. There are >100 ABC transporter predictions in the Strongylocentrotus purpuratus genome, including 40 annotated ABCB, ABCC, and ABCG "multidrug efflux" transporters. Despite the importance of multidrug transporters for protection and signaling, their expression patterns have not been characterized in deuterostome embryos.
Sea urchin embryos expressed 20 ABCB, ABCC, and ABCG transporter genes in the first 58 hr of development, from unfertilized egg to early prism. We quantified transcripts of ABCB1a, ABCB4a, ABCC1, ABCC5a, ABCC9a, and ABCG2b, and found that ABCB1a mRNA was 10-100 times more abundant than other transporter mRNAs. In situ hybridization showed ABCB1a was expressed ubiquitously in embryos, while ABCC5a was restricted to secondary mesenchyme cells and their precursors. Fluorescent protein fusions showed localization of ABCB1a on apical cell surfaces, and ABCC5a on basolateral surfaces.
Embryos use many ABC transporters with predicted functions in cell signaling, lysosomal and mitochondrial homeostasis, potassium channel regulation, pigmentation, and xenobiotic efflux. Detailed characterization of ABCB1a and ABCC5a revealed that they have different temporal and spatial gene expression profiles and protein localization patterns that correlate to their predicted functions in protection and development, respectively.
三磷酸腺苷结合盒(ABC)转运蛋白是调节多种化合物和离子细胞内浓度的膜蛋白。在秀丽隐杆线虫基因组中预测有>100 种 ABC 转运蛋白,包括 40 种注释的 ABCB、ABCC 和 ABCG“多药外排”转运蛋白。尽管多药转运蛋白对保护和信号转导很重要,但它们在后口动物胚胎中的表达模式尚未得到描述。
海胆胚胎在发育的前 58 小时内表达了 20 种 ABCB、ABCC 和 ABCG 转运蛋白基因,从受精卵到早期棱柱期。我们定量了 ABCB1a、ABCB4a、ABCC1、ABCC5a、ABCC9a 和 ABCG2b 的转录物,发现 ABCB1a mRNA 的丰度是其他转运蛋白 mRNA 的 10-100 倍。原位杂交显示 ABCB1a 在胚胎中广泛表达,而 ABCC5a 局限于次级间充质细胞及其前体细胞。荧光蛋白融合显示 ABCB1a 定位于细胞表面的顶端,ABCC5a 定位于基底外侧表面。
胚胎使用许多 ABC 转运蛋白,其预测功能涉及细胞信号转导、溶酶体和线粒体稳态、钾通道调节、色素沉着和外来物外排。对 ABCB1a 和 ABCC5a 的详细特征分析表明,它们具有不同的时空基因表达谱和蛋白定位模式,分别与它们在保护和发育中的预测功能相关。