Visualizing Corticotropin-Releasing Hormone Receptor Type 1 Expression and Neuronal Connectivities in the Mouse Using a Novel Multifunctional Allele
Visualizing Corticotropin-Releasing Hormone Receptor Type 1 Expression and Neuronal Connectivities in the Mouse Using a Novel Multifunctional Allele
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DOI：
10.1002/cne.23082
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发表时间：
2012-10-01
影响因子：
2.5
通讯作者：
Deussing, Jan M.
中科院分区：
文献类型：
--
作者：
Kuehne, Claudia;Puk, Oliver;Deussing, Jan M.
促肾上腺皮质激素释放激素（CRH）及其1型受体（CRHR 1）在协调应激的内分泌、自主神经和行为反应中起着重要作用。从功能上剖析CRH/CRHR 1系统的复杂性的先决条件是解开CRHR 1表达神经元及其连接的身份。因此，我们使用敲入方法用tau-lacZ（tZ）报告基因遗传标记CRHR 1表达细胞。表达β-半乳糖苷酶的神经元在脑中的分布和标记的相对强度与先前描述的Crhr 1 mRNA表达完全一致。结合微管结合特性的TAU与Cre-loxP系统允许直接的b-半乳糖苷酶的近端树突，特别是轴突。因此，我们能够可视化CRHR 1神经元的投射，例如纹状体的多巴胺能和多巴胺能传入连接以及位于其斑块隔室内的GABA能CRHR 1表达神经元。此外，tZ报告基因揭示了CRHR 1在脊髓、皮肤和眼睛中表达的新细节。CRHR 1在视网膜中的表达促使CRHR 1与视觉系统相关的新生理作用的鉴定。除了其报告特性之外，这种新的CRHR 1等位基因还包括分别通过Flp和Cre重组酶有条件地恢复或缺失CRHR 1的可能性。最后，该等位基因适合于通过重组酶介导的盒交换进一步操作CRHR 1基因座。总之，这种新的小鼠等位基因将显着促进CRHR 1电路的神经解剖学分析，并开辟了新的途径，以解决CRHR 1功能更详细。神经学比较杂志520：3150-3180，2012. (C)2012 Wiley Periodicals，Inc.
The corticotropin-releasing hormone (CRH) and its type 1 receptor (CRHR1) play a central role in coordinating the endocrine, autonomic, and behavioral responses to stress. A prerequisite to functionally dissect the complexity of the CRH/CRHR1 system is to unravel the identity of CRHR1-expressing neurons and their connectivities. Therefore, we used a knockin approach to genetically label CRHR1-expressing cells with a tau-lacZ (tZ) reporter gene. The distribution of neurons expressing beta-galactosidase in the brain and the relative intensity of labeling is in full accordance with previously described Crhr1 mRNA expression. Combining the microtubule-binding properties of TAU with the Cre-loxP system allowed to direct the b-galactosidase to proximal dendrites, and in particular to axons. Thereby, we were able to visualize projections of CRHR1 neurons such as glutamatergic and dopaminergic afferent connections of the striatum and GABAergic CRHR1-expressing neurons located within its patch compartment. In addition, the tZ reporter gene revealed novel details of CRHR1 expression in the spinal cord, skin, and eye. CRHR1 expression in the retina prompted the identification of a new physiological role of CRHR1 related to the visual system. Besides its reporter properties, this novel CRHR1 allele comprises the possibility to conditionally restore or delete CRHR1 via Flp and Cre recombinase, respectively. Finally, the allele is suitable for further manipulations of the CRHR1 locus by recombinase-mediated cassette exchange. Taken together, this novel mouse allele will significantly facilitate the neuroanatomical analysis of CRHR1 circuits and opens up new avenues to address CRHR1 function in more detail. J. Comp. Neurol. 520:3150-3180, 2012. (C) 2012 Wiley Periodicals, Inc.