小鼠内侧缰核-棘间核通路中富含α3β4烟碱乙酰胆碱受体的突触外表达-动脉网

Abundant extrasynaptic expression of α3β4-containing nicotinic acetylcholine receptors in the medial habenula–interpeduncular nucleus pathway in mice

Nature 等信源发布 2024-06-20 18:28

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AbstractNicotinic acetylcholine receptors (nAChRs) in the medial habenula (MHb)–interpeduncular nucleus (IPN) pathway play critical roles in nicotine-related behaviors. This pathway is particularly enriched in nAChR α3 and β4 subunits, both of which are genetically linked to nicotine dependence. However, the cellular and subcellular expression of endogenous α3β4-containing nAChRs remains largely unknown because specific antibodies and appropriate detection methods were unavailable.

摘要内侧缰核（MHb）-髓间核（IPN）通路中的烟碱乙酰胆碱受体（nAChRs）在尼古丁相关行为中起关键作用。该途径特别富含nAChRα3和β4亚基，这两个亚基在遗传上都与尼古丁依赖有关。然而，内源性含α3β4的nAChRs的细胞和亚细胞表达仍然很大程度上未知，因为无法获得特异性抗体和适当的检测方法。

Here, we successfully uncovered the expression of endogenous nAChRs containing α3 and β4 subunits in the MHb–IPN pathway using novel specific antibodies and a fixative glyoxal that enables simultaneous detection of synaptic and extrasynaptic molecules. Immunofluorescence and immunoelectron microscopy revealed that both subunits were predominantly localized to the extrasynaptic cell surface of somatodendritic and axonal compartments of MHb neurons but not at their synaptic junctions.

在这里，我们使用新型特异性抗体和能够同时检测突触和突触外分子的固定剂乙二醛，成功地发现了MHb-IPN途径中含有α3和β4亚基的内源性nAChRs的表达。免疫荧光和免疫电子显微镜显示，这两个亚基主要定位于MHb神经元的体树突状和轴突区室的突触外细胞表面，但不位于其突触连接处。

Immunolabeling for α3 and β4 subunits disappeared in α5β4-knockout brains, which we used as negative controls. The enriched and diffuse extrasynaptic expression along the MHb–IPN pathway suggests that α3β4-containing nAChRs may enhance the excitability of MHb neurons and neurotransmitter release from their presynaptic terminals in the IPN.

在我们用作阴性对照的α5β4基因敲除大脑中，α3和β4亚基的免疫标记消失了。沿着MHb-IPN途径富集和扩散的突触外表达表明，含有α3β4的nAChRs可能会增强MHb神经元的兴奋性，并增强IPN突触前末端神经递质的释放。

The revealed distribution pattern provides a molecular and anatomical basis for understanding the functional role of α3β4-containing nAChRs in the crucial pathway of nicotine dependence..

揭示的分布模式为理解含α3β4的nAChRs在尼古丁依赖的关键途径中的功能作用提供了分子和解剖学基础。。

IntroductionNicotinic acetylcholine receptors (nAChRs) are widely distributed in the peripheral and central nervous systems and are involved in diverse brain functions. Upon binding the endogenous neurotransmitter acetylcholine or nicotine from smoking, nAChRs allow cations to flow through their channels, thereby modulating neuronal excitability and synaptic transmission1.

简介烟碱型乙酰胆碱受体（nAChRs）广泛分布于外周和中枢神经系统，参与多种脑功能。在结合吸烟产生的内源性神经递质乙酰胆碱或尼古丁后，nAChRs允许阳离子流过其通道，从而调节神经元兴奋性和突触传递1。

nAChRs in the nervous system consist of eight α subunits (α2–7, α9, α10) and three β subunits (β2–4). The specific stoichiometry of α and β subunits confers distinct pharmacological and biophysical properties to nAChRs, including differences in sensitivity to endogenous and exogenous ligands, as well as in the kinetics and amplitude of ion currents through the channels2.

神经系统中的nAChRs由八个α亚基（α2-7，α9，α10）和三个β亚基（β2-4）组成。α和β亚基的特定化学计量赋予nAChRs不同的药理学和生物物理特性，包括对内源性和外源性配体的敏感性差异，以及通过通道的离子电流的动力学和幅度2。

In addition, each subunit is expressed in distinct but overlapping patterns in many cell types throughout the brain3,4, suggesting specific roles for nAChR subtypes in different brain regions.While most nAChRs are characterized by rapid desensitization, α3β4-containing receptors show exceptionally slow desensitization5 and function as the primary receptors in smokers with chronically high blood nicotine levels6.

此外，每个亚基在整个大脑的许多细胞类型中以不同但重叠的模式表达3,4，表明nAChR亚型在不同大脑区域的特定作用。虽然大多数nAChRs的特征是快速脱敏，但含有α3β4的受体表现出异常缓慢的脱敏5，并且是长期高血尼古丁水平吸烟者的主要受体6。

Genome-wide association studies show that single-nucleotide polymorphisms in the CHRNΑ5-CHRNΑ3-CHRNΒ4 gene cluster, which encodes α5, α3, and β4 nAChR subunits, reduce the aversive properties of nicotine and heighten the risk of excessive smoking7,8,9. Overexpression of the β4 subunit in the medial habenula–interpeduncular nucleus (MHb–IPN) pathway augments α3β4 receptor-mediated currents and increases aversion to nicotine, while a decrease in these currents attenuates nicotine aversion10.

。内侧缰核-髓间核（MHb-IPN）途径中β4亚基的过表达增加了α3β4受体介导的电流并增加了对尼古丁的厌恶，而这些电流的减少则减弱了尼古丁的厌恶10。

Conversely, viral-mediated α3 knockdown in the rat MHb or IPN increases nicotine intake11. These studies collectively suggest that the norm.

相反，大鼠MHb或IPN中病毒介导的α3敲低会增加尼古丁摄入量11。这些研究共同表明，规范。

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Download referencesFundingThis study was supported by Ministry of Education,Culture,Sports,Science and Technology (17KK0160, 21K06746, 22K06784, 20H05628, 20H05628).Author informationAuthor notesThese authors contributed equally: Asuka Tsuzuki and Miwako Yamasaki.Authors and AffiliationsDepartment of Anatomy, Graduate School of Medicine, Hokkaido University, Sapporo, 060-8638, JapanAsuka TsuzukiDepartment of Anatomy, Faculty of Medicine, Hokkaido University, Sapporo, 060-8638, JapanMiwako Yamasaki, Kohtarou Konno & Masahiko WatanabeDepartment of Functioning and Disability, Faculty of Health Sciences, Hokkaido University, Sapporo, 060-8638, JapanTaisuke MiyazakiInstitute for Animal Experimentation, Faculty of Medicine, Hokkaido University, Sapporo, 060-8638, JapanNorio TakeiDepartment of Cellular and Molecular Physiology, Department of Neuroscience, and Kavli Institute for Neuroscience, Yale University School of Medicine, New Haven, CT, 06520, USASusumu TomitaDepartment of Physiology, School of Medicine, Keio University, Tokyo, 160-8582, JapanMichisuke YuzakiAuthorsAsuka TsuzukiView author publicationsYou can also search for this author in.

下载参考文献资助这项研究得到了教育，文化，体育，科学和技术部的支持（17KK016021K067462K0678420H0562820H05628）。作者信息作者注意到这些作者做出了同样的贡献：Asuka Tsuzuki和Miwako Yamasaki。作者和所属机构北海道大学医学研究生院解剖学系，札幌，060-8638，日本北海道大学医学院解剖学系，札幌，060-8638，日本山崎和子，Kohtarou Konno＆Masahiko WatanabeDepartment of Functional and Disability，Faculty of Health Sciences，北海道大学，札幌，060-8638，日本北海道大学医学院宫崎骏动物实验研究所，札幌，060-8638，日本Orio TakeiDepartment of Cellular and Molecular Physiology，Department of Neuroscience，and Kavli Institute for Neuroscience，Yale University School of Medicine，New Haven，CT，06520，USASusumu TomitaDepartment of Physiology，School of Medicine，Keio University，Tokyo，160-8582，JapanMichisuke YuzakiAuthorsAsuka TsuzukiView author Publications你也可以在中搜索这位作者。

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PubMed Google ScholarContributionsM.Yamasaki and M.W. designed the research, A.T., M. Yamasaki, K.K., N.T., and M.W. performed the research, S.T., T.M., and M. Yuzaki contributed unpublished reagents/analytic tools, A.T. and M. Yamasaki analyzed the data, and M. Yamasaki and M.W.

PubMed谷歌学术贡献。山崎和M.W.设计了这项研究，A.T.，M.Yamasaki，K.K.，N.T。和M.W.进行了研究，S.T.，T.M。和M.Yuzaki贡献了未发表的试剂/分析工具，A.T.和M.Yamasaki分析了数据，M.Yamasaki和M.W。

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Reprints and permissionsAbout this articleCite this articleTsuzuki, A., Yamasaki, M., Konno, K. et al. Abundant extrasynaptic expression of α3β4-containing nicotinic acetylcholine receptors in the medial habenula–interpeduncular nucleus pathway in mice.

转载和许可本文引用本文Tsuzuki，A.，Yamasaki，M.，Konno，K。等人。小鼠内侧缰核-髓间核通路中含有α3β4的烟碱型乙酰胆碱受体的大量突触外表达。

Sci Rep 14, 14193 (2024). https://doi.org/10.1038/s41598-024-65076-3Download citationReceived: 15 March 2024Accepted: 17 June 2024Published: 20 June 2024DOI: https://doi.org/10.1038/s41598-024-65076-3Share this articleAnyone you share the following link with will be able to read this content:Get shareable linkSorry, a shareable link is not currently available for this article.Copy to clipboard.

Sci Rep 14193（2024）。https://doi.org/10.1038/s41598-024-65076-3Download引文接收日期：2024年3月15日接受日期：2024年6月17日发布日期：2024年6月20日OI：https://doi.org/10.1038/s41598-024-65076-3Share本文与您共享以下链接的任何人都可以阅读此内容：获取可共享链接对不起，本文目前没有可共享的链接。复制到剪贴板。

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