生态HIV脑感染的老鼠模型
Özet
在这里，我们提出了一个协议，建立一个新的大鼠模式，使用有幻想的艾滋病毒（EcoHIV），这是关键，以提高我们对HIV-1病毒库在大脑中的理解，并提供一个系统，以研究艾滋病毒相关的神经认知障碍和相关的合并症（即药物滥用）。
Abstract
据充分研究，EcoHIV感染小鼠模型在研究艾滋病毒相关神经并发症方面具有重大效用。建立EcoHIV感染大鼠模型，用于研究药物滥用和神经认知障碍，将有利于神经HIV和HIV-1相关神经认知障碍（HAND）的研究。在本研究中，我们演示了使用有幻想的艾滋病毒（EcoHIV）成功创建了活性艾滋病毒感染的老鼠模型。首先，EcoHIV 的扁桃体结构在培养的 293 FT 单元中包装了 48 小时。然后，有条件的介质被集中和滴答作响。接下来，我们将EcoHIV-EGFP的双边立体毒性注射到F344/N大鼠脑组织中。感染一周后，在受感染的脑组织中检测到EGFP荧光信号，表明EcoHIV成功地诱发大鼠的艾滋病毒感染。此外，还对微胶质细胞标记物Ibba1进行了免疫染色。结果表明，微胶质是窝藏EcoHIV的主要细胞类型。此外，EcoHIV大鼠在时间处理方面表现出改变，这是HAND潜在的神经行为机制，感染后8周出现突触功能障碍。本研究将EcoHIV艾滋病毒-1感染模型共同扩展至大鼠，为研究大脑中的HIV-1病毒库以及手部和药物滥用等相关合并症提供了宝贵的生物系统。
Introduction
生物系统增强了我们对HIV-1相关神经认知障碍（HAND）及其基础神经机制2的理解。确定哪个生物系统最适合任何特定的研究，往往取决于利益问题2。宿主动物模型范围的限制对艾滋病毒-1疾病发展的研究提出了挑战。为了研究HIV-1病毒的复制和发病机理，Potash等人3日创建了活性HIV-1感染的小鼠模型，用生态MLV gp80取代了HIV表面包络糖蛋白gp120的编码区域，从而成功地在4号小鼠身上进行了病毒复制。在有幻想的HIV（EcoHIV）小鼠注射尾静脉后，观察到许多特征与HIV-1血清阳性个体（如受感染的淋巴细胞和巨噬细胞，针对抗病毒免疫反应，以及炎症3，5，6）相似。
虽然老鼠和老鼠都是Muridae的成员，但基本物种的差异可能会影响它们适合特定的实验问题7。因此，将EcoHIV感染模型推广到大鼠（通常用于药物滥用和神经认知障碍的研究）将有利于神经HIV的研究。例如，其较大的尺寸使朱拉导管植入药物自我管理程序更实用8。大鼠的药物自我管理技术被用来评估艾滋病毒-19的动机。此外，许多神经认知/行为任务最初是为10号大鼠设计的。在这里，我们报告利用大鼠EcoHIV的立体毒性注射来扩展EcoHIV感染模式，并提供了一个关键的机会来解决与神经HIV和HAND相关的新问题。
Protocol
Representative Results
Discussion
在本议定书中，我们建立了大鼠感染EcoHIV的艾滋病毒模式。具体来说，我们描述了EcoHIV的双边立体毒性注射到皮层，在注射后7天成功地诱导了大鼠大脑中的活性HIV感染。此外，我们证明，大鼠的EcoHIV感染可能是研究HAND关键方面的一个很好的生物系统。在EcoHIV感染后八周，大鼠表现出严重的神经认知障碍，其中包括在NAc的MSN中时间处理和突触功能障碍的改变。鉴于动物模型对神经HIV和<sup class="xref"…
Açıklamalar
The authors have nothing to disclose.
Acknowledgements
这项工作由NIH赠款HD043680、MH106392、DA013137和NS100624资助。
Materials
| 293FT cells | ThermoFisher Scientific | R70007 | |
| Antibiotic-Antimycotic solution | Cellgro | 30004CI | 100X |
| Corning BioCoatGelatin 75cm² Rectangular Canted Neck Cell Culture Flask with Vented Cap | Life Technologies | 354488 | |
| Corning DMEM with L-Glutamine, 4.5 g/L Glucose and Sodium Pyruvate | Life Technologies | 10013CV | |
| Cover glass | VWR | 637-137 | |
| drill | |||
| Dumont #5 Forceps | World Precision Instruments | 14095 | |
| Dumont #7 Forceps | World Precision Instruments | 14097 | |
| Eppendorf Snap-Cap Microcentrifuge Biopur Safe-Lock Tubes | Life Technologies | 22600028 | |
| Ethicon Vicryl Plus Antibacterial, 4-0 Polyglactin 910 Suture, 27in. FS-2 | Med Vet International | VCP422H | |
| Hamilton syringe | Hamilton | 1701 | |
| Invitrogen Lipofectamine 3000 Transfection Reagent | Life Technologies | L3000015 | |
| Iris Forceps | World Precision Instruments | 15914 | |
| Iris Scissors | World Precision Instruments | 500216 | |
| Lentivirus-Associated p24 ELISA Kit | Cell Biolabs, inc. | VPK-107-5 | |
| Lenti-X Concentrator | Takara | PT4421-2 | |
| Opti-MEM I Reduced Serum Medium | Life Technologies | 11058021 | |
| Paraformaldehyde | Sigma-Aldrich | 158127-500G | |
| Paraformaldehyde | Sigma | P6148 | |
| ProLong Gold | Fisher Scientific | P36930 | |
| Sevoflurane | Merritt Veterinary Supply | 347075 | |
| stereotaxic apparatus | Kopf Instruments | Model 900 | |
| SuperFrost Plus Slides | Fisher Scientific | 12-550-154% | |
| Vannas Scissors | World Precision Instruments | 500086 |
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