通过一些逆行示踪剂标记从剑突核出现的血清素能神经元回路。

IF 2 3区 工程技术 Q2 ANATOMY & MORPHOLOGY
Mona N Hussein
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引用次数: 0

摘要

羟色胺(5-羟色胺,5-HT)是一种非常重要的神经递质,它从剑突核进入多个脑区。血清素能神经元连接在大脑中具有多种功能。在这项研究中,我们使用了多种技术来追踪背侧饶舌(DR)和正中饶舌(MnR)中向下丘脑弓状核(Arc)投射的血清素能神经元、下丘脑背内侧核(DM)、下丘脑外侧区(LH)、下丘脑室旁核(PVH)、下丘脑腹内侧核(VMH)、下丘脑筋膜(FC)和下丘脑内侧核(MHb)。本研究使用了霍乱毒素亚单位 B(CTB)、逆转录腺相关病毒(rAAV-CMV-mCherry)、去糖蛋白狂犬病病毒(RV-ΔG),以及在 C57BL/6 小鼠中同时显微注射 rAAV2-retro-Cre-tagBFP 和 AAV-dio-mCherry。此外,还将 rAAV2-retro-Cre-tagBFP 显微注射到 Ai9 小鼠体内。利用羟色胺免疫组化技术检测了逆行追踪的剑突核中的羟色胺能神经元。结果表明,在Ai9小鼠体内注射rAAV2-retro-Cre-tagBFP是追踪血清素能神经元回路的最佳方法。前面列出的所有核团都有来自DR和MnR的5-羟色胺能神经元投射,只有FC例外,它很少有来自DR的投射。5-羟色胺能神经元的投射通过弧下三叶核(SPTg)指向弧。此外,RV-ΔG示踪剂显示,来自DR的单突触非羟色胺能神经元投射指向弧。此外,rAAV示踪剂揭示了从剑突核指向弧的单突触血清素能神经元连接。这些发现验证了几种逆行描记剂在神经趋向性方面的差异。继续发现几种新的血清素能神经回路对于未来发现这些回路的功能至关重要。研究亮点:向 C57BL/6 和 Ai9 小鼠显微注射各种逆行描记剂。在 Ai9 小鼠中显微注射 rAAV2-retro-Cre-tagBFP 是表征血清素能神经元回路的最佳方法。DR、MnR和SPTg核向下丘脑弓状核发出单突触血清素能神经元投射。rAAV2-retro-Cre-tagBFP 显微注射 Arc、DM、LH 和 VMH 后,不同脑核中逆行标记神经元的全脑定量分析。图中显示了向 PVH、DM、LH、Arc、VMH、MHb 和 FC 方向出现的正中和背侧饶弓血清素能神经元的差异定量分析。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Labeling of the serotonergic neuronal circuits emerging from the raphe nuclei via some retrograde tracers.

Serotonin (5-hydroxytryptamine, 5-HT) is a very important neurotransmitter emerging from the raphe nuclei to several brain regions. Serotonergic neuronal connectivity has multiple functions in the brain. In this study, several techniques were used to trace serotonergic neurons in the dorsal raphe (DR) and median raphe (MnR) that project toward the arcuate nucleus of the hypothalamus (Arc), dorsomedial hypothalamic nucleus (DM), lateral hypothalamic area (LH), paraventricular hypothalamic nucleus (PVH), ventromedial hypothalamic nucleus (VMH), fasciola cinereum (FC), and medial habenular nucleus (MHb). Cholera toxin subunit B (CTB), retro-adeno-associated virus (rAAV-CMV-mCherry), glycoprotein-deleted rabies virus (RV-ΔG), and simultaneous microinjection of rAAV2-retro-Cre-tagBFP with AAV-dio-mCherry in C57BL/6 mice were used in this study. In addition, rAAV2-retro-Cre-tagBFP was microinjected into Ai9 mice. Serotonin immunohistochemistry was used for the detection of retrogradely traced serotonergic neurons in the raphe nuclei. The results indicated that rAAV2-retro-Cre-tagBFP microinjection in Ai9 mice was the best method for tracing serotonergic neuron circuits. All of the previously listed nuclei exhibited serotonergic neuronal projections from the DR and MnR, with the exception of the FC, which had very few projections from the DR. The serotonergic neuronal projections were directed toward the Arc by the subpeduncular tegmental (SPTg) nuclei. Moreover, the RV-ΔG tracer revealed monosynaptic non-serotonergic neuronal projections from the DR that were directed toward the Arc. Furthermore, rAAV tracers revealed monosynaptic serotonergic neuronal connections from the raphe nuclei toward Arc. These findings validate the variations in neurotropism among several retrograde tracers. The continued discovery of several novel serotonergic neural circuits is crucial for the future discovery of the functions of these circuits. RESEARCH HIGHLIGHTS: Various kinds of retrograde tracers were microinjected into C57BL/6 and Ai9 mice. The optimum method for characterizing serotonergic neuronal circuits is rAAV2-retro-Cre-tagBFP microinjection in Ai9 mice. The DR, MnR, and SPTg nuclei send monosynaptic serotonergic neuronal projections toward the arcuate nucleus of the hypothalamus. Whole-brain quantification analysis of retrograde-labeled neurons in different brain nuclei following rAAV2-retro-Cre-tagBFP microinjection in the Arc, DM, LH, and VMH is shown. Differential quantitative analysis of median and dorsal raphe serotonergic neurons emerging toward the PVH, DM, LH, Arc, VMH, MHb, and FC is shown.

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来源期刊
Microscopy Research and Technique
Microscopy Research and Technique 医学-解剖学与形态学
CiteScore
5.30
自引率
20.00%
发文量
233
审稿时长
4.7 months
期刊介绍: Microscopy Research and Technique (MRT) publishes articles on all aspects of advanced microscopy original architecture and methodologies with applications in the biological, clinical, chemical, and materials sciences. Original basic and applied research as well as technical papers dealing with the various subsets of microscopy are encouraged. MRT is the right form for those developing new microscopy methods or using the microscope to answer key questions in basic and applied research.
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