Phospholipase D (PLD) hydrolyzes phosphatidylcholine to generate phosphatidic acid and choline. Studies in cultured cells and Drosophila melanogaster have implicated PLD in the regulation of many cellular functions, including intracellular vesicle trafficking, cell proliferation and differentiation. However, the function of PLD in vertebrate development has not been explored. Here we report cloning and characterization of a zebrafish PLD1 (pld1) homolog. Like mammalian PLDs, zebrafish Pld1 contains two conservative HKD motifs. Maternally contributed pld1 transcripts are uniformly distributed in early embryo. Localized expression of pld1 is observed in the notochord during early segmentation, in the somites during later segmentation and in the liver at the larval stages. Studies in intact and cell-free preparations demonstrate evolutionary conservation of regulation. Inhibition of Pld1 expression using antisense morpholino oligonucleotides (MO) interfering with the translation or splicing of pld1 impaired intersegmental vessel (ISV) development. Incubating embryos with 1-butanol, which diverts production of phosphatidic acid to a phosphatidylalcohol, caused similar ISV defects. To determine where Pld1 is required for ISV development we performed transplantation experiments. Analyses of the mosaic Pld1 deficient embryos showed partial suppression of ISV defects in the segments containing transplanted wild-type notochord cells but not in the ones containing wild-type somitic cells. These results provide the first evidence that function of Pld1 in the developing notochord is essential for vascular development in vertebrates.

中文翻译：

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斑马鱼节间血管的血管生成需要磷脂酶 D1。

磷脂酶 D (PLD) 水解磷脂酰胆碱以生成磷脂酸和胆碱。对培养细胞和黑腹果蝇的研究表明 PLD 参与了许多细胞功能的调节，包括细胞内囊泡运输、细胞增殖和分化。然而，PLD 在脊椎动物发育中的功能尚未被探索。在这里，我们报告了斑马鱼 PLD1 (pld1) 同源物的克隆和表征。与哺乳动物 PLD 一样，斑马鱼 Pld1 包含两个保守的 HKD 基序。母体贡献的 pld1 转录本均匀分布在早期胚胎中。pld1 的局部表达在早期分割期间在脊索中、在后期分割期间在体节中以及在幼虫阶段在肝脏中观察到。对完整和无细胞制剂的研究证明了调节的进化保守性。使用干扰 pld1 的翻译或剪接的反义吗啉代寡核苷酸 (MO) 抑制 Pld1 表达会损害节间血管 (ISV) 的发育。将胚胎与 1-丁醇一起孵化，将磷脂酸的产生转化为磷脂醇，会导致类似的 ISV 缺陷。为了确定 ISV 开发所需的 Pld1，我们进行了移植实验。对嵌合 Pld1 缺陷胚胎的分析表明，在含有移植的野生型脊索细胞的片段中部分抑制了 ISV 缺陷，但在含有野生型体细胞的片段中则没有。