Unexpected BrdU inhibition on astrocyte-to-neuron conversion  

作　　者：Tao Wang Jian-Cheng Liao Xu Wang Qing-Song Wang Kai-Ying Wan Yi-Yi Yang Qing He Jia-Xuan Zhang Gong Chen Wen Li 

机构地区：[1]Guangdong-Hong Kong-Macao Institute of CNS Regeneration(GHMICR),Jinan University,Guangzhou,Guangdong Province,China [2]Department of Neurosurgery,The First Affiliated Hospital of Jinan University,Guangzhou,Guangdong Province,China

出　　处：《Neural Regeneration Research》2022年第7期1526-1534,共9页中国神经再生研究（英文版）

基　　金：supported by the Natural Science Foundation of Guangdong Province of China,Nos.2021A1515011237(to WL),2020A1515010854(to QSW);the National Natural Science Foundation of China,Nos.U1801681(to GC),31701291(to WL);the Guangdong Province Science and Technology Planning Project of China,No.2018B030332001(to GC)。

摘　　要：5-Bromo-2′-deoxyuridine(BrdU)is a halogenated pyrimidine that can be incorporated into newly synthesized DNA during the S phase of the cell cycle.BrdU is widely used in fate-mapping studies of embryonic and adult neurogenesis to identify newborn neurons,however side effects on neural stem cells and their progeny have been reported.In vivo astrocyte-to-neuron(AtN)conversion is a new approach for generating newborn neurons by directly converting endogenous astrocytes into neurons.The BrdU-labeling strategy has been used to trace astrocyte-converted neurons,but whether BrdU has any effect on the AtN conversion is unknown.Here,while conducting a NeuroD1-mediated AtN conversion study using BrdU to label dividing reactive astrocytes following ischemic injury,we accidentally discovered that BrdU inhibited AtN conversion.We initially found a gradual reduction in BrdU-labeled astrocytes during NeuroD1-mediated AtN conversion in the mouse cortex.Although most NeuroD1-infected astrocytes were converted into neurons,the number of BrdU-labeled neurons was surprisingly low.To exclude the possibility that this BrdU inhibition was caused by the ischemic injury,we conducted an in vitro AtN conversion study by overexpressing NeuroD1 in cultured cortical astrocytes in the presence or absence of BrdU.Surprisingly,we also found a significantly lower conversion rate and a smaller number of converted neurons in the BrdU-treated group compared with the untreated group.These results revealed an unexpected inhibitory effect of BrdU on AtN conversion,suggesting more caution is needed when using BrdU in AtN conversion studies and in data interpretation.

关 键 词：5-bromo-2′-deoxyuridine NeuroD1 astrocyte-to-neuron conversion reprogramming neural regeneration reactive astrocytes neurons lineage tracing fate mapping neural stem cell 

分 类 号：R741.02[医药卫生—神经病学与精神病学]