文章摘要
斑马鱼nos2a基因的生长调控作用
Role of nos2a in regulating the growth of zebrafish (Danio rerio)
投稿时间:2020-07-07  修订日期:2020-12-28
DOI:
中文关键词: 一氧化氮合酶  斑马鱼  基因敲除  生长激素  生长
英文关键词: nitric oxide synthase  zebrafish  knockout  growth hormone  growth
基金项目:国家自然科学基金项目(面上项目,重点项目,重大项目)
作者单位邮编
高磊 教育部水生遗传资源开发利用重点实验室 201306
黄亚娟 教育部水生遗传资源开发利用重点实验室 201306
Rachit Penglee 教育部水生遗传资源开发利用重点实验室 201306
易欣鑫 教育部水生遗传资源开发利用重点实验室 201306
鲍宝龙 教育部水生遗传资源开发利用重点实验室 201306
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中文摘要:
      鱼类的生长调控一直是水产领域研究的热点。为了在斑马鱼中探索诱导型一氧化氮合酶(Nos2)在生长调控中的作用,我们通过CRISPR/Cas9基因编辑技术,首次构建了可稳定遗传的nos2a缺失型(nos2a-/-)斑马鱼家系。通过监测其生长过程发现,nos2a-/- 斑马鱼的整体生长趋势较nos2a正常型(nos2a+/+)斑马鱼偏低,且nos2a-/-成鱼的体型显著小于同期的nos2a+/+ 斑马鱼(P <0.01):nos2a+/+成鱼体长(29.53 ± 0.60) mm,nos2a-/-成鱼体长(26.34 ± 0.69) mm;nos2a+/+成鱼体质量(512.8 ± 30.75) mg,nos2a-/-成鱼体质量(394.4 ± 16.91) mg。而在肌纤维密度(P =0.718)及躯椎骨的横截面积上(P =0.548)两者没有显著差异。qPCR检测发现,出膜8天时,nos2a-/- 斑马鱼的生长激素基因gh1的mRNA水平(P <0.01)及类胰岛素生长因子基因igf1的mRNA水平(P <0.001)均显著低于nos2a+/+ 斑马鱼;出膜68天时,nos2a-/- 斑马鱼的gh1 (P <0.0001)和igf1 (P <0.0001)的mRNA水平同样显著低于nos2a+/+ 斑马鱼。以上结果表明,斑马鱼中nos2a的功能缺失导致鱼体生长受到抑制,而gh1及igf1 mRNA的水平下调是其潜在原因。
英文摘要:
      To explore the role of inducible nitric oxide synthase (Nos2) on fish growth regulation, we generated nos2a deficient (nos2a-/-) zebrafish using CRISPR/Cas9 technology for the first time. By monitoring their growth process, we found the growth trend of nos2a-/- zebrafish was slower than nos2a+/+ zebrafish, and the adult body size of nos2a-/- zebrafish was smaller than that of nos2a+/+ zebrafish (P<0.01): the mean length and weight of nos2a+/+ zebrafish were 29.53 mm and 512.8 mg, respectively, that of nos2a-/- zebrafish was 26.34 mm and 394.4 mg. Additionally, there is no notable difference in the density of muscle fibers (P=0.718) and sectional area of vertebrae (P=0.548) between nos2a+/+ and nos2a-/- zebrafish. Moreover, compared with nos2a+/+ zebrafish at 8 dph (days after hatching), the mRNA level of growth hormone (gh1, P <0.01) and insulin-like growth factor-1 (igf1, P <0.001) of nos2a-/- zebrafish at 8 dph was lower significantly; and it is the same as that at 68 dph: gh1, P <0.0001; igf1, P <0.0001. These findings indicate that the defect of nos2a negatively affected zebrafish growth, and the reduced gene mRNA level of growth hormone was the potential factor.
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