Silencing of Syntaxin 1A in the Dopaminergic Neurons Decreases the Activity of the Dopamine Transporter and Prevents Amphetamine-Induced Behaviors in C. elegans.
Silencing of Syntaxin 1A in the Dopaminergic Neurons Decreases the Activity of the Dopamine Transporter and Prevents Amphetamine-Induced Behaviors in C. elegans.
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DOI：
10.3389/fphys.2018.00576
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发表时间：
2018
影响因子：
4
通讯作者：
Carvelli L
中科院分区：
文献类型：
--
作者：
Lanzo A;Safratowich BD;Kudumala SR;Gallotta I;Zampi G;Di Schiavi E;Carvelli L
多巴胺转运蛋白（DAT）是一种细胞膜蛋白，其主要功能是将突触间隙释放的多巴胺（DA）再摄取回多巴胺能神经元。先前的研究表明，DAT的活性受变构蛋白（如Syntaxin-1A）的调节，并被滥用药物（如安非他明（Amph））改变。由于秀丽隐杆线虫同时表达DAT （DAT-1）和Syntaxin-1A (UNC-64)，我们使用该模型系统来研究UNC-64在培养的多巴胺能神经元和活体动物中缺乏表达所引起的功能和行为影响。使用一种可遗传的RNA沉默技术，我们能够特异性地敲除多巴胺能神经元中的unc-64。这种细胞特异性敲除方法避免了unc-64敲除突变引起的多效性表型，并确保了多巴胺能特异性unc-64沉默传递给后代。我们发现，与dat-1敲除和dat-1沉默系相似，多巴胺能神经元中unc-64表达减少的动物在运动行为测试中对Amph治疗没有反应。我们的体外数据表明，在unc-64沉默动物的神经元培养物中，与对照组相比，DA摄取减少了30%，这种减少与从dat-1沉默动物分离的神经元中测量到的减少相似（40%）。此外，减少unc-64在多巴胺能神经元中的表达显著减少Amph引起的DA释放。由于在线虫中DAT-1是唯一能够再摄取DA的蛋白，这些数据表明，多巴胺能神经元中unc-64的表达减少会降低DAT重新积累突触DA的能力。此外，这些结果表明，unc-64在多巴胺能神经元中的表达减少，消除了Amph诱导的运动行为。综上所述，这些数据表明Syntaxin-1A在Amph引起的功能和行为影响中都起着重要作用。
The dopamine transporter (DAT) is a cell membrane protein whose main function is to reuptake the dopamine (DA) released in the synaptic cleft back into the dopaminergic neurons. Previous studies suggested that the activity of DAT is regulated by allosteric proteins such as Syntaxin-1A and is altered by drugs of abuse such as amphetamine (Amph). Because Caenorhabditis elegans expresses both DAT (DAT-1) and Syntaxin-1A (UNC-64), we used this model system to investigate the functional and behavioral effects caused by lack of expression of unc-64 in cultured dopaminergic neurons and in living animals. Using an inheritable RNA silencing technique, we were able to knockdown unc-64 specifically in the dopaminergic neurons. This cell-specific knockdown approach avoids the pleiotropic phenotypes caused by knockout mutations of unc-64 and ensures the transmission of dopaminergic specific unc-64 silencing to the progeny. We found that, similarly to dat-1 knockouts and dat-1 silenced lines, animals with reduced unc-64 expression in the dopaminergic neurons did not respond to Amph treatment when tested for locomotor behaviors. Our in vitro data demonstrated that in neuronal cultures derived from animals silenced for unc-64, the DA uptake was reduced by 30% when compared to controls, and this reduction was similar to that measured in neurons isolated from animals silenced for dat-1 (40%). Moreover, reduced expression of unc-64 in the dopaminergic neurons significantly reduced the DA release elicited by Amph. Because in C. elegans DAT-1 is the only protein capable to reuptake DA, these data show that reduced expression of unc-64 in the dopaminergic neurons decreases the capability of DAT in re-accumulating synaptic DA. Moreover, these results demonstrate that decreased expression of unc-64 in the dopaminergic neurons abrogates the locomotor behavior induced by Amph. Taken together these data suggest that Syntaxin-1A plays an important role in both functional and behavioral effects caused by Amph.
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影响因子：
3.3
作者：
Cervinski, M. A.;Foster, J. D.;Vaughan, R. A.
通讯作者：
Vaughan, R. A.
影响因子：
3.5
作者：
Gallotta I;Mazzarella N;Donato A;Esposito A;Chaplin JC;Castro S;Zampi G;Battaglia GS;Hilliard MA;Bazzicalupo P;Di Schiavi E
通讯作者：
Di Schiavi E
影响因子：
16.2
作者：
Christensen, M;Estevez, A;Strange, K
通讯作者：
Strange, K
影响因子：
3.4
作者：
Illiano, Placido;Lanzo, Ambra;Di Schiavi, Elia
通讯作者：
Di Schiavi, Elia
影响因子：
11.1
作者：
Cartier, Etienne;Hamilton, Peter J.;Belovich, Andrea N.;Shekar, Aparna;Campbell, Nicholas G.;Saunders, Christine;Andreassen, Thorvald F.;Gether, Ulrik;Veenstra-Vanderweele, Jeremy;Sutcliffe, James S.;Ulery-Reynolds, Paula G.;Erreger, Kevin;Matthies, Heinrich J. G.;Galli, Aurelio
通讯作者：
Galli, Aurelio