Transduction profiles of recombinant adeno-associated virus vectors derived from serotypes 2 and 5 in the nigrostriatal system of rats
Transduction profiles of recombinant adeno-associated virus vectors derived from serotypes 2 and 5 in the nigrostriatal system of rats
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DOI：
10.1128/jvi.78.13.6808-6817.2004
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发表时间：
2004-07-01
影响因子：
5.4
通讯作者：
Büeler, H
中科院分区：
文献类型：
--
作者：
Paterna, JC;Feldon, J;Büeler, H
我们比较了来自血清型2和血清型5（分别为rAAV-2和rAAV-5）的滴度匹配重组腺相关病毒（rAAV）在大鼠黑质纹状体系统中的转导效率和倾向。两种血清型（表达增强型绿色荧光蛋白[EGFP]）通过立体定向手术进入相同动物的纹状体（STR）、黑质（SN）或内侧前脑束（MFB）的不同半球。虽然两种血清型都能有效地在STR内转导神经元，但rAAV-5导致的egfp表达面积比rAAV-2大得多。然而，用rAAV-2载体转导的神经元表达更高水平的EGFP。与此结果一致的是，在纹状体注入rAAV-5后，从转导的纹状体神经元发出的EGFP阳性投射覆盖了更大的网状神经中枢（SNr）区域，但在纹状体注入rAAV-2后，SNr纤维中的EGFP水平更高。我们还比较了两种载体的逆行转导潜力，发现纹状体递送rAAV-5导致SN致密部和腹侧被盖区内多巴胺能细胞体的转导明显增加。同样，egfp转导的纹状体神经元仅在rAAV-5的神经传递后被检测到。此外，我们证明纹状体AAV-5载体递送后，转导谱稳定长达9个月。最后，虽然我们没有直接靶向海马，但在将rAAV-5（而不是rAAV-2）递送到MFB后，观察到海马神经元的有效和广泛的转导。
We compared the transduction efficiencies and tropisms of titer-matched recombinant adeno-associated viruses (rAAV) derived from serotypes 2 and 5 (rAAV-2 and rAAV-5, respectively) within the rat nigrostriatal system. The two serotypes (expressing enhanced green fluorescent protein [EGFP]) were delivered by stereotaxic surgery into the same animals but different hemispheres of the striatum (STR), the substantia nigra (SN), or the medial forebrain bundle (MFB). While both serotypes transduced neurons effectively within the STR, rAAV-5 resulted in a much larger EGFP-expressing area than did rAAV-2. However, neurons transduced with rAAV-2 vectors expressed higher levels of EGFP. Consistent with this result, EGFP-positive projections emanating from transduced striatal neurons covered a larger area of the SN pars reticulata (SNr) after striatal delivery of rAAV-5, but EGFP levels in fibers of the SNr were higher after striatal injection of rAAV-2. We also compared the potentials of the two vectors for retrograde transduction and found that striatal delivery of rAAV-5 resulted in significantly more transduced dopaminergic cell bodies within the SN pars compacta and ventral tegmental area. Similarly, EGFP-transduced striatal neurons were detected only after nigral delivery of rAAV-5. Furthermore, we demonstrate that after striatal AAV-5 vector delivery, the transduction profiles were stable for as long as 9 months. Finally, although we did not target the hippocampus directly, efficient and widespread transduction of hippocampal neurons was observed after delivery of rAAV-5, but not rAAV-2, into the MFB.