自由活动小鼠使用遥测长期血压测量
Summary
The goal of this protocol is to assess systemic blood pressure in conscious freely moving mice using implantable radio-telemetry devices.
Abstract
中的新血管活性药物的发展，动脉血压监测是用于评价新提出的药物的功效的关键。事实上，研究集中在新的潜在的治疗靶标使用基因改变的小鼠中发现需要全身动脉压力变化的可靠，长期评估。目前，金标准获得门诊老鼠血压长期测量采用植入无线电发射器，这就需要动脉插管。这种技术消除了对束缚，抑制，或麻醉的数据采样期间引入应力和工件的动物。然而，在经由导管小鼠动脉血压监测可以是相当有挑战性由于动脉的小尺寸。在这里，我们提出一个一步一步的指导，说明了无线电发射器和颈动脉C的成功皮下植入的关键钥匙通道环中的小鼠。我们还包括了一周期手术后恢复的后自由移动小鼠的长期血压活性的实例。根据这一程序将允许同时从多个可靠的动物直接血压记录。
Introduction
高血压是心血管疾病的主要危险因素之一，它无疑是一个重大的公共卫生问题无论是在发达国家和发展中国家1。实验性高血压几种动物模型已经发展到模仿像那些在人体2观察到的高血压反应。其中，门诊鼠标代表了一个很好的模型来研究的起源与高血压，允许在长期暴露于高血压的后果体内分析的进展。
血压（BP）的小鼠监测已经帮助研究人员解开参与生理学和疾病，如高血压和心脏衰竭3,4病理生理学几种机制。事实上，小鼠基因组的操纵允许代转基因或基因靶向模型合适的用于研究高血压5,6。然而，即使是温和的操作清醒小鼠的诱导兴奋，能够在数据采集过程可能引进假象，而使用镇静剂或镇静剂的深刻影响血压7。这些方面是特别重要的，必须在考虑尝试长期血压监测时作出。
有几种方法在小鼠中以记录血压，和目前最常用的技术之间的比较已在别处讨论8,9。然而，对于测量血压的AHA建议的结论是，BP的动脉内测量通常是优选的，因为它们直接测量血压在一段10长时间的能力。加之动脉压的直接测量无线电遥测是监测生理功能清醒和自由活动的实验动物，同时尽量减少压力和环保相关的人为9，11国家的最先进的方法。无线电遥测提供自动收集血压，心脏速率，体温和动物活动来自多个清醒动物的能力。
虽然这种方法已成为很受欢迎，在许多实验室，在小鼠无线电遥测植入可以技术上具有挑战性。这里，我们显示了一步一步的协议，阐释如何植入在小鼠一压力传感器。该技术涉及在小鼠体内的探针的皮下插入，引导导管到颈部并通过左侧颈动脉附近的主动脉弓转发。远程捕获的数据显示活在电脑显示器上。数据也存储为“离线”的分析。
Protocol
Representative Results
Discussion
植入无线电遥测已经在过去十年显著改善;较小的探针尺寸使得所述植入物为动物创伤小，延长电池寿命有助于降低成本，并独立遥测频率消除接收器之间的串扰。遥测被认为是国家的最先进的方法为自由移动的动物而不使用约束，人际交往，或者麻醉由其他技术8,9所需的相关文物收集的各种生理参数。
然而，他的一些技术的缺点的用仪器和软件的初始成本相关联。?…
Declarações
The authors have nothing to disclose.
Acknowledgements
This work was supported by the National Institutes of Health (NIH/NHLBI) [Grant no. HL114869] and the support from UTHSC to SM.
Materials
| Small animal anesthesia system | Kent Scientific Corp, Torrington, Connecticut, USA | low-Flow small animal anesthesia system | |
| Pad warmer and mouse termometer | Kent Scientific Corp, Torrington, Connecticut, USA | allows monitoring body temperature, and homeothermic control in small animals. | |
| Binocular Microscope | Kent Scientific Corp, Torrington, Connecticut, USA | KSCXTS-1121 | binocular body with boom stand pole and top LED. |
| Hemostat Forceps | Kent Scientific Corp, Torrington, Connecticut, USA | INS750451 | used to clamp blood vessels or tag sutures |
| Small metal Clips, 7mm, Stainless Steel | Kent Scientific Corp, Torrington, Connecticut, USA | INS750344 | used for skin closure |
| Betadine solution | Purdue Products L.P., Stamford, CT, USA | NDC-67618-150-01 | 10% povidone iodine topical solution |
| Normal saline solution | Abott Laboratories | 04930-04-10 | Needed for preventing tissue from drying. |
| Nair (Hair remover lotion) | Needed for fur removal from the site of incision/surgery. | ||
| Braide silk suture | Teleflex Medical OEM, Coventry, Connecticut, USA | Size 5.0, 6.0, 7.0 | |
| Ethanol | 2716 | 70% ethanol for disinfection | |
| Spring scissors | Fine Science Tool | 15000-10 | for minor dissection |
| Scissors (angled to side) | Fine Science Tool | 14063-011 | No. 3 handle |
| Scalpel blade | 2976-0 | No. 10 | |
| Forceps (curved) | Fine Science Tool | 11150-10 | for holding tissue |
| Forceps (straight) | Fine Science Tool | 11151-10 | for holding tissue |
| Needle holder | Fine Science Tool | 12002-12 | for suturing |
| Fine needle nose Forceps | Fine Science Tool | ||
| Isoflurane | Henry Schein Animal Health, Melville, New York, USA | a general inhalation anesthetic agent | |
| Sterilizer | Benchmark Scientific, 116 Corporate Blvd, South Plainfield, NJ, USA | B1000 | sterilize surgical tools in 5-10 seconds using infrared heating |
| Gauze Pads | Johnson & Johnson, New Brunswick, NJ, USA | JJ8513 | to use for wound cleaning, prepping, scrubbin or dressing. |
| Telemetry Device | Data Sciences International, St. Paul, MN, USA | DSI-PA-C10 | to record blood pressure in freely moving mice |
| Telemetry receiver system coumpled with a PC | Data Sciences International, St. Paul, MN, USA | ||
| small Tubing | Instech Laboratories, Plymouth Meeting, PA USA | BTPE-90 | |
| Vessel Cannulation Forceps, 13cm, 0.5mm OD | World Precision Instruments | 503374 | special vessel cannulation forceps. |
| Tissue adhesive | 3M Animal Care Products, St. Paul, MN, USA | NAC No.: 11380041 | use to close minor wounds, often eliminating the need for sutures and/or bandages |
| Weighing scale | BB300 | Precision Analytical Laboratory Balance |
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