牛奶收集方法小鼠和里夫斯'麂鹿
Summary
Milk collection from animal models facilitates various research avenues: understanding passive immunity, identifying pathogens responsible for vertical transmission and, through the use of transgenic mice, even commercial production of proteins found in human breast milk. Here we illustrate a simple method for milk collection in mice and Reeves’ muntjac deer.
Abstract
Animal models are commonly used throughout research laboratories to accomplish what would normally be considered impractical in a pathogen’s native host. Milk collection from animals allows scientists the opportunity to study many aspects of reproduction including vertical transmission, passive immunity, mammary gland biology, and lactation. Obtaining adequate volumes of milk for these studies is a challenging task, especially from small animal models. Here we illustrate an inexpensive and facile method for milk collection in mice and Reeves’ muntjac deer that does not require specialized equipment or extensive training. This particular method requires two researchers: one to express the milk and to stabilize the animal, and one to collect the milk in an appropriate container from either a Muntjac or mouse model. The mouse model also requires the use of a P-200 pipetman and corresponding pipette tips. While this method is low cost and relatively easy to perform, researchers should be advised that anesthetizing the animal is required for optimal milk collection.
Introduction
动物模型中提供的洞察，不能由体外分析来获得病的病理。提供最有效的结果，以使用密切相关的疾病和感兴趣物种的动物模型是重要的。例如，在穿过了麂（ 麂 ），小亚鹿1,2，和转基因小鼠中表达鹿科朊病毒蛋白（CerTgPrP）3的有用的动物模型鹿科物种。这两个物种是polyestrous，让常年繁殖，因此一致的来源与妊娠有关的组织和体液中，研究具体的机制在生物学鹿科。牛奶有研究的完成在动物模型中比在人类中应用较为简单（且便宜）繁多。研究人员可以调查乳和初乳为1）感染性疾病传播的潜在来源，2）在开发板的由母亲传递给后代的免疫球蛋白被动免疫4和3）乳铁蛋白pment，参与被动免疫的研究人员目前正在试图人类母乳中发现的一种蛋白质商业化生产5。
由小动物收集的牛奶大量可以证明是一项艰巨的任务。罗杰斯提出了一个方法，从大鼠6，随后在小鼠用于收集牛奶。 DePeters和霍维提出了两种方法，奶收集，一种使用由连接到巴斯德吸管的橡胶吸管灯泡产生的手动产生的真空，并且第二要求挤奶单元，然后将其连接到一个真空源（建造这样作为一个水龙头）收获鼠标奶7。在这里，我们提出了一个简单的，低成本的方法从小鼠和里夫斯'麂鹿，只需要现成的实验室设备和基本的技术技能收集牛奶。我们的方法产生足量的牛奶各种应用程序。
Protocol
Representative Results
Discussion
鼠标
有几个因素来自鼠标收集牛奶时要考虑到，包括1）的时间预留收集，催产素2）的给药剂量，3）有多少幼仔大坝是目前护理和4）金额自分娩已经通过在收集时的时间。使用以往的研究为指导，我们开始着手优化产奶量的条件。
此前有研究直接相关的产奶量与产仔8。不幸的是，产仔数是不能被控制的变量。然而，挤奶后的分娩?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
These experiments were funded by the NIH grant #RAI093634A. We express immense gratitude to Jenny Powers, DVM, PhD, for her assistance in muntjac milking.
Materials
| Materials-Mouse | |||
| Name of Material/Reagent | Vendor | Catalog Number | Comments |
| Ketamine (KetaVed 100 mg/mL) (dose: 80-100 mg/kg) | MWI/Vedco | 011075 | |
| Xylazine (TranquiVed 100 mg/ml) (dose:5-10 mg/kg) | MWI/Vedco | NADA 139236 | |
| Eye lubricant | MWI/Apexa | 701013 | |
| Oxytocin (2 IU/kg) | Vet One | 501013 | |
| Materials- Muntjac | |||
| Name of Material/Reagent | Vendor | Catalog Number | Comments |
| Midazolam (1-2 mg/kg) | Amerisource Bergen Drug Corp | 924326 | optional: researcher can either use Midazolam on its own or BAM combination as an anesthetic |
| Butorphanol tartrate (Torbugesic) (0.45 mg/kg) | Fort Dodge Animal Health | NADA 135-780 | |
| Azaperone tartrate (0.25 mg/kg) | ZooPharm | ||
| Medetomadine HCl (0.07 mg/kg) | ZooPharm | ||
| Antipamezol hydrochloride (Antisedan) (dose: 0.25 mg/kg) | Orion Corporation | NADA 141-033 | |
| Oxytocin (10 IU/kg) | Vet One | 501013 | |
| Equipment- Mouse | |||
| Name of Equipment | Vendor | Catalog Number | Comments |
| P-200 Pipetman | Gilson | F123601 | |
| 200 μL pipette tips | Light Labs | B-2004-SH | |
| 1 ml syringes | MWI | 005427 | |
| 27 gauge needles | MWI | 002031 | |
| Equipment- Muntjac | |||
| Name of Equipment | Vendor | Catalog Number | Comments |
| 15 ml conical tubes | VWR | 21008-918 | |
| 22 gauge needles | MWI | 873 | |
| 3 ml luer lock syringes | MWI | 001377 |
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