乳腺的发育和功能在小鼠模型的评价
Summary
这种方法介绍如何剖析和评估小鼠乳腺发育和功能。切除乳腺评估使用全挂载的发展程度，而牛奶弹射使用催产素为基础的肌上皮细胞收缩测定评价。
Abstract
15-20叶组成的人体乳腺分泌乳汁，到乳头管系统开放的一个分支。这些叶是自己的终末导管小叶分泌肺泡和融合的管道1单位组成。在小鼠实验中，观察到类似的架构在怀孕管道和肺泡内的结缔组织基质中穿插。小鼠乳腺上皮细胞是像由两层细胞，管腔细胞基底膜2的范围表示的肌上皮细胞的外层包围的内层组成的导管系统树。出生时，只有残留的导管树目前，主风管和15-20分支机构组成。伸长率和放大分公司开始在青春期的开始，4周龄左右，3,4,5激素的影响下。在10个星期，大部分基质入侵的管道由一个复杂的系统，将发生在每个发情周期，直到怀孕 2分支和回归周期。在怀孕开始，开始第二阶段的发展，与上皮细胞的增殖和分化形成葡萄状的牛奶分泌结构称为肺泡 6,7的。整个分娩和哺乳，牛奶是生产管腔分泌细胞，并储存在肺泡腔内。催产素的释放，仔兔哺乳引起的神经反射刺激，诱导肺泡周围和沿导管的肌上皮细胞的同步收缩，让牛奶通过管道输送到乳头它成为可用的幼崽 8 。乳腺的发育，分化和功能的紧密配合和要求，不仅基质和上皮细胞之间的相互作用，而且之间的肌上皮细胞和管腔内的上皮细胞9,10,11。从而，在这些相互作用中有牵连的许多基因的突变可能会损害或者在妊娠晚期和分泌的激活 ，导致泌乳12,13在青春期或肺泡在怀孕早期，分化形成导管伸长。在这篇文章中，我们描述了如何解剖小鼠乳腺和评估他们的发展，使用整个坐骑。我们还演示了如何评价肌上皮收缩和牛奶弹射使用前体内催产素的功能测定。对乳腺的发育和功能的基因突变的影响，因此可以在原地决定执行这两种技术在突变体和野生型对照组小鼠。
Protocol
Discussion
乳腺发育和功能的紧密协调。突变小鼠中可能隐藏的基因突变，会损害乳腺的发育和功能，突出了需要评估腺架构13,12。本文中描述的乳腺发育的各个阶段，整个安装可以执行。通常情况下，上皮细胞的发展是评估在青春期开始（〜4周龄），青春期（〜6周龄）中，进入青春期后（约10-12周龄）2,10 。在这几个阶段，管分支的数量，可确定的末端芽和管道的长度以量化程度的</sup…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这些研究是由CIHR CBCRA补助金，以DWL。知识产权是由CIHR – STP，FRSQ和CIHR奖学金资助。 MKGS是由OGS和CIHR – STP奖学金。作者感谢协助他用鼠标育种凯文巴尔。
Materials
| Reagent | Company | Catalogue number |
|---|---|---|
| Carmine | Sigma-Aldrich | C1022 |
| Aluminum potassium sulfate | Sigma-Aldrich | A6435 |
| Thymol | Sigma-Aldrich | T0501 |
| Methyl salicylate | Sigma-Aldrich | M6752 |
| Oxytocin | Sigma-Aldrich | O3251 |
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