Neural Regen Res. 2020 Oct 9
Zongqin Xiang, Liang Xu, Minhui Liu, Qingsong Wang, Wen Li, Wenliang Lei, and Gong Chen
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Research Field: CNS

Keywords: adeno-associated viruses, astrocyte, dosage, glia-to-neuron conversion, in vivo reprogramming, lineage tracing, neuron, retrovirus

AAV Serotype: AAV9 and AAV5

Dose: 1 μL of virus mixture AAV9 hGFAP::Cre (1 × 10^10 gc/mL) and FLEX-CAG::mCherry (1 × 10^12 gc/mL) was injected at the same coordinates. For intact mouse cortex, 1 μL of retroviruses CAG::NeuroD1- IRES-GFP (1 × 107 TU/mL) or 1 μL of AAV5 GFAP::NeuroD1 (1 × 1012 gc/mL) were injected at the similar coordinates described above.

Routes of Administration: intraperitoneal injection

Targeted organ: brain


Regenerating functional new neurons in the adult mammalian central nervous system has been proven to be very challenging due to the inability of neurons to divide and repopulate themselves after neuronal loss. Glial cells, on the other hand, can divide and repopulate themselves under injury or diseased conditions. We have previously reported that ectopic expression of NeuroD1 in dividing glial cells can directly convert them into neurons. Here, using astrocytic lineage-tracing reporter mice (Aldh1l1-CreERT2 mice crossing with Ai14 mice), we demonstrate that lineage-traced astrocytes can be successfully converted into NeuN-positive neurons after expressing NeuroD1 through adeno-associated viruses. Retroviral expression of NeuroD1 further confirms that dividing glial cells can be converted into neurons. Importantly, we demonstrate that for in vivo cell conversion study, using a safe level of adeno-associated virus dosage (1010–1012 gc/mL, 1 µL) in the rodent brain is critical to avoid artifacts caused by toxic dosage, such as that used in a recent bioRxiv study (2 × 1013 gc/mL, 1 µL, mouse cortex). For therapeutic purpose under injury or diseased conditions, or for non-human primate studies, adeno-associated virus dosage needs to be optimized through a series of dose-finding experiments. Moreover, for future in vivo glia-to-neuron conversion studies, we recommend that the adeno-associated virus results are further verified with retroviruses that mainly express transgenes in dividing glial cells in order to draw solid conclusions. The study was approved by the Laboratory Animal Ethics Committee of Jinan University, China (approval No. IACUC-20180330-06) on March 30, 2018.

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