Recombinase-independent AAV for anterograde transsynaptic tracing.

IF 4.3 3区材料科学 Q1 ENGINEERING, ELECTRICAL & ELECTRONIC ACS Applied Electronic Materials Pub Date : 2023-09-15 DOI:10.1186/s13041-023-01053-7

Islam Faress, Valentina Khalil, Haruka Yamamoto, Szilard Sajgo, Keisuke Yonehara, Sadegh Nabavi

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Abstract

Viral transsynaptic labeling has become indispensable for investigating the functional connectivity of neural circuits in the mammalian brain. Adeno-associated virus serotype 1 (AAV1) allows for anterograde transneuronal labeling and manipulation of postsynaptic neurons. However, it is limited to delivering an AAV1 expressing a recombinase which relies on using transgenic animals or genetic access to postsynaptic neurons. We reasoned that a strong expression level could overcome this limitation. To this end, we used a self-complementary AAV of serotype 1 (scAAV1) under a strong promoter (CAG). We demonstrated the anterograde transneuronal efficiency of scAAV1 by delivering a fluorescent marker in mouse retina-superior colliculus and thalamic-amygdala pathways in a recombinase-independent manner in the mouse brain. In addition to investigating neuronal connectivity, anterograde transsynaptic AAVs with a strong promoter may be suitable for functional mapping and imaging.

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重组非依赖性AAV用于顺行经突触追踪。

病毒经突触标记对于研究哺乳动物大脑中神经回路的功能连接已变得不可或缺。腺相关病毒血清型1（AAV1）允许顺行跨神经元标记和突触后神经元的操作。然而，它仅限于递送表达重组酶的AAV1，该重组酶依赖于使用转基因动物或遗传途径进入突触后神经元。我们推断，强大的表达水平可以克服这一限制。为此，我们在强启动子（CAG）下使用了血清型1的自互补AAV（scAAV1）。我们通过在小鼠大脑中以重组酶无关的方式在小鼠视网膜上丘和丘脑杏仁核通路中递送荧光标记物，证明了scAAV1的顺行跨神经元效率。除了研究神经元连接外，具有强启动子的顺行突触性AAV可能适用于功能定位和成像。

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