Diversity of Reporter Expression Patterns in Transgenic Mouse Lines Targeting Corticotropin-Releasing Hormone-Expressing Neurons
Diversity of Reporter Expression Patterns in Transgenic Mouse Lines Targeting Corticotropin-Releasing Hormone-Expressing Neurons
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DOI：
10.1210/en.2015-1673
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发表时间：
2015-12-01
期刊：
影响因子：
4.8
通讯作者：
Baram, Tallie Z.
中科院分区：
文献类型：
--
作者：
Chen, Yuncai;Molet, Jenny;Baram, Tallie Z.
转基因小鼠，包括针对促肾上腺皮质激素释放因子（CRF或CRH）的品系，已被广泛用于研究应激神经生物学。这些强大的工具有望彻底改变我们对CRH表达神经元的定位和连接以及CRH在正常和病理条件下的关键作用的理解。使用细胞类型特异性转基因小鼠的研究的准确解释至关重要地依赖于内源性肽和报告基因的表达之间的一致性。如果报告基因表达不能忠实地再现天然基因表达，那么操纵无意靶向细胞的效果可能被错误归因。在这里，我们研究了CRH和报告基因表达模式在3个成年转基因小鼠：Crh-IRES-Cre; Ai 14（tdTomato小鼠），Crfp3.0 CreGFP和Crh-GFP BAC。我们使用由Vale产生的CRH抗血清，在使用CRH缺失小鼠验证其特异性后。我们的分析集中在应激显著区域，包括下丘脑、杏仁核、终纹床核和海马。内源性CRH的表达模式在野生型和转基因小鼠中是一致的。在tdTomato小鼠中，大多数表达CRH的神经元共表达报告基因，但报告基因在海马CA 1和CA 3中发现了一些不表达CRH的神经元样细胞。在Crfp 3.0 CreGFP小鼠中，在中央杏仁核中发现CRH和报告基因的共表达，在其他评估区域中不太常见。在Crh-GFP BAC小鼠中，绝大多数神经元表达CRH或报告基因，几乎没有重叠。这些数据突出了在3个可用的转基因小鼠中报告基因和内源性CRH的一致表达的显著多样性。这些发现应该有助于解释从使用这些强大的神经生物学工具中出现的重要科学发现。
Transgenic mice, including lines targeting corticotropin-releasing factor (CRF or CRH), have been extensively employed to study stress neurobiology. These powerful tools are poised to revolutionize our understanding of the localization and connectivity of CRH-expressing neurons, and the crucial roles of CRH in normal and pathological conditions. Accurate interpretation of studies using cell type-specific transgenic mice vitally depends on congruence between expression of the endogenous peptide and reporter. If reporter expression does not faithfully reproduce native gene expression, then effects of manipulating unintentionally targeted cells may be misattributed. Here, we studied CRH and reporter expression patterns in 3 adult transgenic mice: Crh-IRES-Cre; Ai14 (tdTomato mouse), Crfp3.0 CreGFP, and Crh-GFP BAC. We employed the CRH antiserum generated by Vale after validating its specificity using CRH-null mice. We focused the analyses on stress-salient regions, including hypothalamus, amygdala, bed nucleus of the stria terminalis, and hippocampus. Expression patterns of endogenous CRH were consistent among wild-type and transgenic mice. In tdTomato mice, most CRH-expressing neurons coexpressed the reporter, yet the reporter identified a few non-CRH-expressing pyramidal-like cells in hippocampal CA1 and CA3. In Crfp 3.0 CreGFP mice, coexpression of CRH and the reporter was found in central amygdala and, less commonly, in other evaluated regions. In Crh-GFP BAC mice, the large majority of neurons expressed either CRH or reporter, with little overlap. These data highlight significant diversity in concordant expression of reporter and endogenous CRH among 3 available transgenic mice. These findings should be instrumental in interpreting important scientific findings emerging from the use of these potent neurobiological tools.