Jiao Shuang, Wu Zhihao, Tan Xungang, Sui Yulei, Wang Lijuan, You Feng
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao, 266071, Shandong, People's Republic of China.
Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, 7 Nanhai Road, Qingdao, 266071, Shandong, People's Republic of China.
Fish Physiol Biochem. 2017 Apr;43(2):385-395. doi: 10.1007/s10695-016-0294-3. Epub 2016 Sep 27.
Although chromosome set manipulation techniques including polyploidy induction and gynogentic induction in flatfish are becoming increasingly mature, there exists a poor understanding of their effects on embryonic development. PAX3 plays crucial roles during embryonic myogenesis and neurogenesis. In olive flounder (Paralichthys olivaceus), there are two duplicated pax3 genes (pax3a, pax3b), and both of them are expressed in the brain and muscle regions with some subtle regional differences. We utilized pax3a and pax3b as indicators to preliminarily investigate whether chromosome set manipulation affects embryonic neurogenesis and myogenesis using whole-mount in situ hybridization. In the polyploid induction groups, 94 % of embryos in the triploid induction group had normal pax3a/3b expression patterns; however, 45 % of embryos in the tetraploid induction group showed abnormal pax3a/3b expression patterns from the tailbud formation stage to the hatching stage. Therefore, the artificial induction of triploidy and tetraploidy had a small or a moderate effect on flounder embryonic myogenesis and neurogenesis, respectively. In the gynogenetic induction groups, 87 % of embryos in the meiogynogenetic diploid induction group showed normal pax3a/3b expression patterns. However, almost 100 % of embryos in the gynogenetic haploid induction group and 63 % of embryos in the mitogynogenetic diploid induction group showed abnormal pax3a/3b expression patterns. Therefore, the induction of gynogenetic haploidy and mitogynogenetic diploidy had large effects on flounder embryonic myogenesis and neurogenesis. In conclusion, the differential expression of pax3a and pax3b may provide new insights for consideration of fish chromosome set manipulation.
尽管包括在比目鱼中诱导多倍体和雌核发育在内的染色体组操作技术日益成熟，但人们对其对胚胎发育的影响仍知之甚少。PAX3在胚胎肌发生和神经发生过程中发挥着关键作用。在牙鲆（Paralichthys olivaceus）中，有两个重复的pax3基因（pax3a、pax3b），它们在脑和肌肉区域均有表达，但存在一些细微的区域差异。我们利用pax3a和pax3b作为指标，通过整体原位杂交初步研究染色体组操作是否会影响胚胎神经发生和肌发生。在多倍体诱导组中，三倍体诱导组94%的胚胎具有正常的pax3a/3b表达模式；然而，四倍体诱导组45%的胚胎从尾芽形成期到孵化期显示出异常的pax3a/3b表达模式。因此，人工诱导三倍体和四倍体分别对比目鱼胚胎肌发生和神经发生有较小或中等程度的影响。在雌核发育诱导组中，减数雌核发育二倍体诱导组87%的胚胎显示出正常的pax3a/3b表达模式。然而，雌核发育单倍体诱导组几乎100%的胚胎和有丝分裂雌核发育二倍体诱导组63%的胚胎显示出异常的pax3a/3b表达模式。因此，诱导雌核发育单倍体和有丝分裂雌核发育二倍体对比目鱼胚胎肌发生和神经发生有较大影响。总之，pax3a和pax3b的差异表达可能为鱼类染色体组操作的研究提供新的思路。
