在新生小鼠静脉注射
Summary
儿科疾病的动物模型可以体验早期发病和严重的疾病进展。临床相关治疗交付给年轻小鼠模型可以在技术上难度。这个协议描述了一种非侵入性的静脉内注射的方法在生命的最初2日龄新生小鼠。
Abstract
静脉注射是一种临床应用的方式提供治疗。对于成年啮齿类动物和大型动物，静脉注射在技术上是可行的程序。然而，一些小鼠模型可以有早发性疾病具有病情进展快，使潜在疗法的管理难度。的时间（或面部）静脉只是前耳芽在小鼠和显然对于前两天可见在出生后的头使用解剖显微镜的任一侧。在此窗口中，颞静脉可以与体积可达50微升注射。注射是安全的，由两个幼仔和堤坝的耐受性良好。一种典型的注射过程中1-2分钟，在此之后，小狗返回到笼中完成。到了第三日龄静脉是很难想象和注射过程在技术上变得不可靠。这一技术已被用于递送的腺相关病毒（AAV）VECT口服补液盐，这反过来又可以用于根据所述病毒血清型所选择的动物的生活提供几乎全身性的，稳定的转基因表达。
Introduction
递送治疗剂到在儿科疾病的鼠模型中的中枢神经系统（CNS）中的仍然是一个挑战。小鼠模型中的新生疾病状态是矮小和发育不成熟，因此可能难以直接注入在中枢神经系统内的适当的结构。血管内注射的治疗剂是一种非侵入性的，良好耐受的方法，提供的细胞，药物，或病毒载体的整个身体，包括中枢神经系统1-5和视网膜3,5-9。以前的出版物描述了使用透10,11颞面部静脉注射，无需解剖镜下11,12，还是需要两个人来注入10。在这个协议中所描述的注射技术是有利的，因为一个人可以注入幼仔，和光源，以查看颞静脉没有触摸小狗，省去了手术带或小狗的连接到一个固定的表面诸如透射11。输送的腺相关病毒载体的血清型9（AAV9）在小鼠中产生强大的表达在神经元和星形胶质细胞的整个大脑和脊髓（ 图1）。血管内递送病毒载体到颞浅面静脉已被可靠地使用在各种研究中新生小鼠治疗小儿神经肌肉病脊髓性肌萎缩（SMA），2,4,13,14，并最终增加了处理的小鼠的寿命。
血管内注射新生小鼠也有效地靶向外周神经系统和外周器官（ 图2）。以下注射的AAV，背根神经节，肝脏，心脏，骨骼肌，肺和肠的肠肌丛的转导已观察1,3,6,7,15。中枢神经系统和周围的广泛传导，使理想的注射这种方法需要对全球的疾病表现转基因，比如高雪氏病16和其他溶酶体贮积病17,18，巴滕病和相关的神经元蜡样lipofuscinoses，19和巴比二氏综合征，一种遗传性多系统疾病发病的早期儿童20出现的症状。血管内注射到新生小鼠中，也应考虑作为新的建模系统范围的儿科疾病的方法。这种技术已被翻译为较大的动物模型5,21和血管内注射已经存在作为递送治疗剂的临床上可接受的方法。
目前的协议描述了通过颞浅静脉脸上交付代理商新生小鼠不晚于产后第2天注射一种简单，有效的方法，可以通过一个单一的完成，实行个人和既受两个幼崽和水坝的耐受性。幼崽体验最小痛苦和快速恢复。 ImportanTLY，成功的注入将导致管理代理的全球交付。这个协议是适当的递送病毒载体，药物制剂或细胞对新生小鼠。
Protocol
Representative Results
Discussion
血管内递送药物到中枢神经系统或整个身体难以在疾病的新生鼠模型。所描述的协议，是静脉内给药溶液加入到新生小鼠以最小的设备要求一种快速，相对非侵入性的方式。虽然颞面部静脉可以通过肉眼观察，注射液可以具有更高的准确性与使用显微镜和光纤光源的，特别是对于一个不熟练的喷射器。在新生小鼠的血管内注射剂具有较高的成功率12，和AAV转导已经在早幼崽被观察到即使注…
Divulgations
The authors have nothing to disclose.
Acknowledgements
作者要感谢SMA的NINDS，FightSMA和家庭提供财政支持。 SEGL由NINDS培训资助＃5T32NS077984-02支持。
Materials
| Name of Material/ Equipment | Company | Catalog Number | Comments/Description |
| Thinpro Insulin Syringe | Terumo | SS30M3009 | 3/10cc, 3/8" needle, 30G, 1 per mouse |
| Evans Blue Dye | Sigma-Aldrich | E2129 | Dilute to 1% with 1X Phosphate Buffered Saline |
| Cotton Tipped Applicators | Fisher Scientific | 23-400-101 | |
| Fiber Optic Light Source | Fisher Scientific | 12-562-36 | |
| Dissecting Microscope |
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