孤独谱系障碍相关基因dia1r斑马鱼突变体的构建及行为学表型探析

Construction of Deleted-in-Autism 1 Related Gene Mutant Zebrafish and Analysis of Its Autism-like Behaviors

dia1r基因是人类孤独症谱系障碍相关基因.临床研究发现,缺失dia1r可能会导致患者面部畸形、智力障碍和语言发育迟缓等症状.目前,对dia1r基因的功能及分子机制知之甚少,仅有少数在鸡胚、斑马鱼、小鼠中的表达定位研究.本文通过CRISPR/Cas9系统构建了dia1r敲除的斑马鱼突变体模型,成功获得了稳定遗传的纯合突变体.对突变体进行行为分析发现,与野生型斑马鱼相比,纯合突变体幼鱼的自发运动能力降低,趋触性减弱,对光暗交替刺激更加敏感.在成鱼社交偏好实验中发现,雄性纯合突变体比野生型表现出较弱的社交偏好.

The human gene dia1r is an Autism Spectrum Disorders(ASDs)-related gene.It is reported that two male siblings with dysmorphic faces,ambiguous genitalia,intellectual disability,and speech delay both carried a hemizygous missense variant in CXorf36(also known as dia1r).At present,only the expression pattern of dia1r has been studied in mice,zebrafish,and chicken embryo,the function of dia1r in vivo is still unclear in any of these model organisms.In this study,dia1r was knocked out in zebrafish using CRISPR/Cas9,and a stable mutant line was successfully obtained.The behavior of homozygous mutant larvae was studied.We found that the locomotor activity of homozygous mutant larvae was decreased,and their anxiety-like behavioral response was decreased in a thigmotaxis assay.We further observed that the homozygous mutant larvae were more sensitive to the alternation of light and darkness.In the social preference assay,male homozygous mutant adults showed weaker social preference than male wildtype adults.