Effects of SHANK2 gene knockdown on early neural development of induced pluripotent stem cell derived neuron
作者单位：Guangdong-Hongkong-Macau Institute of CNS Regeneration Joint International Research Laboratory of CNS Regeneration Ministry of Education of PRC Jinan University Co-innovation Center of Neuroregeneration Nantong University
会议名称：《中国神经科学学会第十二届全国学术会议》
会议日期：2017年
学科分类：1001[医学-基础医学(可授医学、理学学位)] 100101[医学-人体解剖与组织胚胎学] 10[医学]
关 键 词：SHANK2 gene Induced pluripotent stem cells Gene knockdown Neuronal morphology
摘 要：OBJECTIVE: To investigate the effect of SHANK2 knockdown on the early morphological development of neurons derived from human induced pluripotent stem cell（iPSCs）. METHODS: Induced pluripotent stem cells（iPSCs） were differentiated into neural progenitor cells（NPC） and NPCs were differentiated into neurons. The models were divided into sh Control group（infected with control Lenti virus packed with a red fluorescent protein expression sequence） and sh SHANK2 group（infected with SHANK2 Lenti virus packed with a red fluorescent protein expression sequence）. The soma area, growth cone area, neurite length and branches number of neurons were analyzed by immunohistochemical method on selected differentiation days（Day3, Day6, Day9, Day12, and Day15）. RESULTS:（1） The model with NPC was generated in vitro significantly.（2） Compared with control, SHANK2 gene knockdown resulted in the decrease of soma area at Day9、Day12、Day15.（3） Compared with control, the number of total branches, primary branches, and primary branches with SHANK2 gene knockdown were reduced, and the length of the neurite, axon, and dendrite were reduced at Day6、Day9、Day12、Day15.（4） Compared with control, SHANK2 gene knockdown resulted in the decrease of three subtypes of growth cone area bluntended（collapsed with no visible filopodia or lamellipodia）, filopodial（growth cones with numerous filopodia and a small or absent lamellipodial veil）, or lamellipodial（well spread growth cones with elaborate lamellipodia） at Day6 、 Day9 、 Day12 、 Day15. CONCLUSION: The early development of neurons were affected by SHANK2 gene knockdown, leading to abnormal neuronal morphology development.