TAF1-gene editing alters the morphology and function of the cerebellum and cerebral cortex
TAF1-gene editing alters the morphology and function of the cerebellum and cerebral cortex
复制标题
TAF1 基因编辑改变小脑和大脑皮层的形态和功能。
DOI：
10.1016/j.nbd.2019.104539
复制
发表时间：
2019-12-01
影响因子：
6.1
通讯作者：
Nelson, Mark A.
中科院分区：
文献类型：
--
作者：
Janakiraman, Udaiyappan;Yu, Jie;Nelson, Mark A.
TAF 1/MRSX 33智力残疾综合征是由TAF 1基因功能丧失突变引起的X连锁疾病。这些突变如何导致畸形、肌张力减退、智力和运动缺陷尚不清楚。具有胚胎靶向TAF 1的小鼠模型已经失败，这可能是由于TAF 1对于生存力是必需的，优先在早期脑发育中表达，并且不耐受突变。新的动物模型是了解神经元病理学的有价值的工具。在这里，我们报告了一种新的TAF 1 ID综合征动物模型的开发和表征，其中TAF 1基因在胚胎大鼠中缺失，使用成簇规则间隔短回文重复序列（CRISPR）相关蛋白9（Cas9）技术和慢病毒转导介导的体细胞脑转基因。对出生后第3天的大鼠幼仔进行脑室内（ICV）注射gRNA对照或gRNA-TAF 1载体。对大鼠进行一系列行为测试，然后在出生后第14天和第35天对大脑进行组织病理学分析。TAF 1编辑的大鼠在新生儿和青少年发育阶段都表现出行为缺陷。TAF 1的缺失导致浦肯野细胞的发育不全和丧失。我们还观察到在TAF 1编辑的动物中小脑颗粒层内GFAP阳性星形胶质细胞减少和Iba 1阳性小胶质细胞增加。免疫染色显示CaV3.1 T型钙通道的表达减少。TAF 1编辑大鼠的异常运动症状与不规则小脑输出有关，这是由于突触前CaV3.1丢失导致浦肯野细胞内在活性变化引起的。该动物模型为研究TAF 1异常相关条件下的神经元功能障碍提供了一个强大的新工具，并应证明可用于开发治疗TAF 1 ID综合征的治疗策略。
TAF1/MRSX33 intellectual disability syndrome is an X-linked disorder caused by loss-of-function mutations in the TAF1 gene. How these mutations cause dysmorphology, hypotonia, intellectual and motor defects is unknown. Mouse models which have embryonically targeted TAF1 have failed, possibly due to TAF1 being essential for viability, preferentially expressed in early brain development, and intolerant of mutation. Novel animal models are valuable tools for understanding neuronal pathology. Here, we report the development and characterization of a novel animal model for TAF1 ID syndrome in which the TAF1 gene is deleted in embryonic rats using clustered regularly interspaced short palindromic repeats (CRISPR) associated protein 9 (Cas9) technology and somatic brain transgenesis mediated by lentiviral transduction. Rat pups, post-natal day 3, were subjected to intracerebroventricular (ICV) injection of either gRNA-control or gRNA-TAF1 vectors. Rats were subjected to a battery of behavioral tests followed by histopathological analyses of brains at post-natal day 14 and day 35. TAF1-edited rats exhibited behavioral deficits at both the neonatal and juvenile stages of development. Deletion of TAF1 lead to a hypoplasia and loss of the Purkinje cells. We also observed a decreased in GFAP positive astrocytes and an increase in Iba1 positive microglia within the granular layer of the cerebellum in TAF1-edited animals. Immunostaining revealed a reduction in the expression of the CaV3.1 T-type calcium channel. Abnormal motor symptoms in TAF1-edited rats were associated with irregular cerebellar output caused by changes in the intrinsic activity of the Purkinje cells due to loss of pre-synaptic CaV3.1. This animal model provides a powerful new tool for studies of neuronal dysfunction in conditions associated with TAF1 abnormalities and should prove useful for developing therapeutic strategies to treat TAF1 ID syndrome.