Department of Neurology, Institute of Neurology of First Affiliated Hospital, Institute of Neuroscience, and Fujian Key Laboratory of Molecular Neurology, Fujian Medical University, Fuzhou, 350005, China.
Mol Brain. 2022 Jul 23;15(1):65. doi: 10.1186/s13041-022-00953-4.
Primary familial brain calcification (PFBC) is a neurogenetic disorder characterized by bilateral calcified deposits in the brain. We previously identified that MYORG as the first pathogenic gene for autosomal recessive PFBC, and established a Myorg-KO mouse model. However, Myorg-KO mice developed brain calcifications until nine months of age, which limits their utility as a facile PFBC model system. Hence, whether there is another typical animal model for mimicking PFBC phenotypes in an early stage still remained unknown. In this study, we profiled the mRNA expression pattern of myorg in zebrafish, and used a morpholino-mediated blocking strategy to knockdown myorg mRNA at splicing and translation initiation levels. We observed multiple calcifications throughout the brain by calcein staining at 2-4 days post-fertilization in myorg-deficient zebrafish, and rescued the calcification phenotype by replenishing myorg cDNA. Overall, we built a novel model for PFBC via knockdown of myorg by antisense oligonucleotides in zebrafish, which could shorten the observation period and replenish the Myorg-KO mouse model phenotype in mechanistic and therapeutic studies.
原发性家族性脑钙化症 (PFBC) 是一种神经遗传性疾病，其特征是大脑双侧有钙化沉积物。我们之前发现 MYORG 是常染色体隐性遗传 PFBC 的第一个致病基因，并建立了 Myorg-KO 小鼠模型。然而，Myorg-KO 小鼠直到 9 个月大时才出现脑钙化，这限制了它们作为简便 PFBC 模型系统的用途。因此，是否存在另一种能够在早期模拟 PFBC 表型的典型动物模型仍不得而知。在这项研究中，我们对斑马鱼中的 myorg mRNA 表达模式进行了分析，并使用基于 morpholino 的阻断策略在剪接和翻译起始水平上敲低 myorg mRNA。我们在受精后 2-4 天通过 calcein 染色观察到 myorg 缺陷型斑马鱼的整个大脑中有多处钙化，并用补充的 myorg cDNA 挽救了钙化表型。总的来说，我们通过在斑马鱼中使用反义寡核苷酸敲低 myorg 建立了一个新的 PFBC 模型，这可以缩短观察期，并在机制和治疗研究中补充 Myorg-KO 小鼠模型的表型。
