Investigating Nrg1 Signaling in the Regenerating Axolotl Spinal Cord Using Multiplexed FISH
Investigating Nrg1 Signaling in the Regenerating Axolotl Spinal Cord Using Multiplexed FISH
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DOI：
10.1002/dneu.22670
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发表时间：
2019-05
影响因子：
3
通讯作者：
P. Freitas;Alexander M. Lovely;James R. Monaghan
中科院分区：
文献类型：
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作者：
P. Freitas;Alexander M. Lovely;James R. Monaghan
切断蝾螈尾巴通过激活内源性干细胞导致功能性脊髓再生。确定控制这些神经干细胞中细胞增殖的信号通路将有助于阐明蝾螈再生能力的机制。在这里，我们表明，神经调节蛋白1（NRG 1）/ErbB 2信号是一个重要的途径，在调节神经干细胞增殖的蝾螈脊髓（Ambystoma mexicanum）。同时本地化的nrg 1 mRNA和Nrg 1蛋白进行了利用杂交链反应荧光原位杂交（FISH）方法在组织切片。多重FISH还允许在单个固定切片上对多种细胞类型进行表型分析，从而表征mRNA表达、蛋白质表达和组织结构。ErbB 2的药理学抑制表明，完整的Nrg 1/ErbB 2信号传导对于成人稳态再生以及损伤诱导的脊髓再生至关重要。总之，我们的研究结果突出了NRG 1/ErbB 2信号通路在美西螈神经干细胞增殖中的重要性。
Amputation of a salamander tail leads to functional spinal cord regeneration through activation of endogenous stem cells. Identifying the signaling pathways that control cell proliferation in these neural stem cells will help elucidate the mechanisms underlying the salamander’s regenerative ability. Here, we show that neuregulin 1 (Nrg1)/ErbB2 signaling is an important pathway in the regulation of neural stem cell proliferation in the spinal cord of the axolotl salamander (Ambystoma mexicanum). Simultaneous localization of nrg1 mRNA and Nrg1 protein was performed by utilizing a hybridization chain reaction fluorescence in situ hybridization (FISH) methodology in tissue sections. Multiplexed FISH also permitted the phenotyping of multiple cell types on a single fixed section allowing the characterization of mRNA expression, protein expression, and tissue architecture. Pharmacological inhibition of ErbB2 showed that intact Nrg1/ErbB2 signaling is critical for adult homeostatic regeneration as well as for injury‐induced spinal cord regeneration. Overall, our results highlight the importance of the NRG1/ErbB2 signaling pathway in neural stem cell proliferation in the axolotl.