A comparison of transduction efficiency of medial septal neurons by adeno-associated viruses with three promoters

IF 2.3 4区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Neuroscience Methods Pub Date : 2025-09-05 DOI:10.1016/j.jneumeth.2025.110570

Alena A. Koryagina , Yulia V. Dobryakova , Konstantin A. Gerasimov , Ghofran Alhalabi , Alexandr A. Moshchenko , Vsevolod V. Belousov , Alexey P. Bolshakov

{"title":"A comparison of transduction efficiency of medial septal neurons by adeno-associated viruses with three promoters","authors":"Alena A. Koryagina , Yulia V. Dobryakova , Konstantin A. Gerasimov , Ghofran Alhalabi , Alexandr A. Moshchenko , Vsevolod V. Belousov , Alexey P. Bolshakov","doi":"10.1016/j.jneumeth.2025.110570","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><div>Most researchers rely on popular promoters like the synthetic CAG promoter or human synapsin promoter to transduce various brain neurons. However, their effectiveness in transducing forebrain cholinergic neurons remains unclear.</div></div><div><h3>New method</h3><div>We compared efficacy of transduction of cholinergic neurons and parvalbumin-positive neurons in the medial septal area of rats and mice by adeno-associated viruses (AAVs) carrying the green fluorescent protein (GFP) marker gene under three distinct promoters—CAG, synapsin, and the mouse choline acetyltransferase (<em>CHAT</em>) promoter.</div></div><div><h3>Results</h3><div>In mice and rats, the CAG and synapsin promoters demonstrated extremely low efficacy for GFP expression in CHAT+ neurons but were effective for transducing PV+ neurons. The <em>CHAT</em> promoter yielded moderate GFP expression in cholinergic neurons in mice, with negligible expression in PV+ neurons, though it also led to off-target expression in other cell types. In rats, the <em>CHAT</em> promoter produced moderate GFP expression in both cholinergic and PV+ neurons; however, the majority of GFP-expressing cells were unrelated to these specific neuronal subtypes.</div><div>Comparison with existing methods: Contrary to expectations, human synapsin and CAG promoters provided negligible expression in cholinergic septal neurons in both rats and mice.</div></div><div><h3>Conclusions</h3><div>The mouse <em>CHAT</em> promoter is significantly more effective for cholinergic neuron expression in rats and mice compared to universal neuronal promoters like CAG and synapsin promoter.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"424 ","pages":"Article 110570"},"PeriodicalIF":2.3000,"publicationDate":"2025-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Neuroscience Methods","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0165027025002146","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}

引用次数: 0

Abstract

Background

Most researchers rely on popular promoters like the synthetic CAG promoter or human synapsin promoter to transduce various brain neurons. However, their effectiveness in transducing forebrain cholinergic neurons remains unclear.

New method

We compared efficacy of transduction of cholinergic neurons and parvalbumin-positive neurons in the medial septal area of rats and mice by adeno-associated viruses (AAVs) carrying the green fluorescent protein (GFP) marker gene under three distinct promoters—CAG, synapsin, and the mouse choline acetyltransferase (CHAT) promoter.

Results

In mice and rats, the CAG and synapsin promoters demonstrated extremely low efficacy for GFP expression in CHAT+ neurons but were effective for transducing PV+ neurons. The CHAT promoter yielded moderate GFP expression in cholinergic neurons in mice, with negligible expression in PV+ neurons, though it also led to off-target expression in other cell types. In rats, the CHAT promoter produced moderate GFP expression in both cholinergic and PV+ neurons; however, the majority of GFP-expressing cells were unrelated to these specific neuronal subtypes.

Comparison with existing methods: Contrary to expectations, human synapsin and CAG promoters provided negligible expression in cholinergic septal neurons in both rats and mice.

Conclusions

The mouse CHAT promoter is significantly more effective for cholinergic neuron expression in rats and mice compared to universal neuronal promoters like CAG and synapsin promoter.

查看原文本刊更多论文

含三种启动子的腺相关病毒对中隔神经元转导效率的NA比较。

背景：大多数研究人员依赖于常用的启动子，如合成CAG启动子或人类突触蛋白启动子来转导各种脑神经元。然而，它们在前脑胆碱能神经元转导中的有效性尚不清楚。新方法：我们比较了携带绿色荧光蛋白（GFP）标记基因的腺相关病毒（aav）在cag、synapsin和小鼠胆碱乙酰转移酶（CHAT）启动子三种不同启动子下对大鼠和小鼠中间隔区胆碱能神经元和小蛋白阳性神经元的转导效果。结果：在小鼠和大鼠中，CAG和突触素启动子对CHAT+神经元中GFP表达的影响极低，但对PV+神经元的转导有效。CHAT启动子在小鼠胆碱能神经元中产生适度的GFP表达，在PV+神经元中的表达可以忽略不计，尽管它在其他细胞类型中也导致脱靶表达。在大鼠中，CHAT启动子在胆碱能神经元和PV+神经元中均产生适度的GFP表达；然而，大多数表达gfp的细胞与这些特定的神经元亚型无关。与现有方法的比较：与预期相反，人类突触素和CAG启动子在大鼠和小鼠的胆碱能间隔神经元中表达可以忽略不计。结论：小鼠CHAT启动子对大鼠和小鼠胆碱能神经元表达的影响明显高于CAG和突触素启动子等通用神经元启动子。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Journal of Neuroscience Methods 医学-神经科学

CiteScore

7.10

自引率

3.30%

发文量

226

审稿时长

52 days

期刊介绍： The Journal of Neuroscience Methods publishes papers that describe new methods that are specifically for neuroscience research conducted in invertebrates, vertebrates or in man. Major methodological improvements or important refinements of established neuroscience methods are also considered for publication. The Journal''s Scope includes all aspects of contemporary neuroscience research, including anatomical, behavioural, biochemical, cellular, computational, molecular, invasive and non-invasive imaging, optogenetic, and physiological research investigations.