A single-cross, RNA interference-based genetic tool for examining the long-term maintenance of homeostatic plasticity.
A single-cross, RNA interference-based genetic tool for examining the long-term maintenance of homeostatic plasticity.
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DOI：
10.3389/fncel.2015.00107
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发表时间：
2015
影响因子：
5.3
通讯作者：
Frank CA
中科院分区：
文献类型：
--
作者：
Brusich DJ;Spring AM;Frank CA
稳态突触可塑性(HSP)帮助神经元和突触维持生理上适当的输出水平。果蝇幼虫神经肌肉接头(NMJ)是研究HSP的有价值的模型。在这里，我们介绍了一种遗传工具，它允许果蝇研究人员通过一个杂交来检查热休克蛋白的终身维持。该工具是一种果蝇种群，它将GAL4/UAS表达系统与基于RNA干扰(RNAi)的谷氨酸受体亚单位基因敲除相结合。有了这个库存，我们发现了关于热休克蛋白维护的重要新信息。我们回答了一个悬而未决的问题，即突触前CaV2型钙通道在NMJ动态平衡中所起的作用。已发表的实验表明，CaV2α1a亚单位基因杂音(Cac)亚型错义突变会损害NMJ的动态平衡可塑性。在这里，我们报告，通过RNAi降低CAC的表达水平不足以损害体内平衡的可塑性。野生型通道的存在似乎支持HSP--即使当总的CaV2功能严重降低时也是如此。我们还进行了基于RNAi和电生理学的筛查，以确定在整个发育过程中维持体内平衡信号所需的新因素。我们发现了果蝇半胱氨酸串蛋白(CSP)和磷脂酶Cβ(Plc21C)的同源物在热休克蛋白中的新作用。我们通过后续的基因测试来确定这些角色的特征。我们讨论了CSP、Plc21C和相关因子如何调节突触前CaV2功能、突触前钙处理或其他信号过程，这些信号过程对于NMJ功能在整个发育过程中的持续稳态调节至关重要。我们的发现扩大了有助于NMJ持久强度的信号通路和过程的范围。
Homeostatic synaptic plasticity (HSP) helps neurons and synapses maintain physiologically appropriate levels of output. The fruit fly Drosophila melanogaster larval neuromuscular junction (NMJ) is a valuable model for studying HSP. Here we introduce a genetic tool that allows fruit fly researchers to examine the lifelong maintenance of HSP with a single cross. The tool is a fruit fly stock that combines the GAL4/UAS expression system with RNA interference (RNAi)-based knock down of a glutamate receptor subunit gene. With this stock, we uncover important new information about the maintenance of HSP. We address an open question about the role that presynaptic CaV2-type Ca2+ channels play in NMJ homeostasis. Published experiments have demonstrated that hypomorphic missense mutations in the CaV2 α1a subunit gene cacophony (cac) can impair homeostatic plasticity at the NMJ. Here we report that reducing cac expression levels by RNAi is not sufficient to impair homeostatic plasticity. The presence of wild-type channels appears to support HSP—even when total CaV2 function is severely reduced. We also conduct an RNAi- and electrophysiology-based screen to identify new factors required for sustained homeostatic signaling throughout development. We uncover novel roles in HSP for Drosophila homologs of Cysteine string protein (CSP) and Phospholipase Cβ (Plc21C). We characterize those roles through follow-up genetic tests. We discuss how CSP, Plc21C, and associated factors could modulate presynaptic CaV2 function, presynaptic Ca2+ handling, or other signaling processes crucial for sustained homeostatic regulation of NMJ function throughout development. Our findings expand the scope of signaling pathways and processes that contribute to the durable strength of the NMJ.
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DOI：
10.1523/jneurosci.5465-11.2012
发表时间：
2012-06-20
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Dickman DK;Tong A;Davis GW
通讯作者：
Davis GW
影响因子：
16.2
作者：
Bergquist, Sharon;Dickman, Dion K.;Davis, Graeme W.
通讯作者：
Davis, Graeme W.
影响因子：
5.3
作者：
Donnelier J;Braun JE
通讯作者：
Braun JE
影响因子：
2.3
作者：
Dawson-Scully, Ken;Lin, Yongqi;Atwood, Harold L.
通讯作者：
Atwood, Harold L.
DOI：
10.1126/science.1179685
发表时间：
2009-11-20
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Dickman DK;Davis GW
通讯作者：
Davis GW