Inhibitory collaterals in genetically identified medium spiny neurons in mouse primary corticostriatal cultures.
Inhibitory collaterals in genetically identified medium spiny neurons in mouse primary corticostriatal cultures.
复制标题
小鼠原代皮质纹状体培养物中基因鉴定的中型多棘神经元的抑制性络脉。
DOI：
10.1002/phy2.164
复制
发表时间：
2013
影响因子：
2.5
通讯作者：
Vicini,Stefano
中科院分区：
文献类型：
--
作者：
Lalchandani,RupaR;Vicini,Stefano
纹状体中棘神经元（MSN）亚型之间的抑制侧枝已被证明对纹状体输出有重要影响。然而，在传统的离体切片记录中，纹状体msn之间的抑制侧支检测率很低，这使得对这些连接的研究具有挑战性。此外，大多数关于MSN抵押品的研究要么是盲目的，要么是在模型中进行的，其中只能区分一种MSN亚型。在这里，我们描述了一个分离培养系统，使用从出生后0天的转基因小鼠身上收获的纹状体和皮质神经元。这些小鼠分别表达tdTomato和多巴胺D1和D2受体启动子下游的增强型绿色荧光蛋白（EGFP），从而允许同时区分两种主要的msn亚型。在体外，这些神经元发育成棘、超极化静息膜电位并表现出上下状态，同时也随着时间的推移保持两种荧光团的表达。使用体外培养14天的全细胞膜片钳记录，我们能够检测到比之前报道的切片记录高得多的抑制性功能突触。这些侧突触释放γ -氨基丁酸（GABA），并形成其他msn的放电模式。尽管减少的体外模型有许多固有的局限性，但这里描述的培养提供了一个独特的机会来研究可识别的msn之间经常观察到的功能附带物。此外，培养的神经元可以控制细胞外环境，有可能研究抑制msn的药理调节。
Inhibitory collaterals between striatal medium spiny neuron (MSN) subtypes have been shown to critically influence striatal output. However, the low rate of inhibitory collateral detection between striatal MSNs in conventional ex vivo slice recordings has made the study of these connections challenging. Furthermore, most studies on MSN collaterals have been conducted either blind or in models, in which only one MSN subtype can be distinguished. Here, we describe a dissociated culture system using striatal and cortical neurons harvested from genetically modified mice at postnatal day 0. These mice express tdTomato and enhanced green fluorescent protein (EGFP) downstream of the dopamine D1 and D2 receptor promoters, respectively, allowing for simultaneous distinction between the two major subtypes of MSNs. In vitro, these neurons develop spines, hyperpolarized resting membrane potentials and exhibit up‐and‐down states, while also maintaining expression of both fluorophores through time. Using paired whole‐cell patch‐clamp recordings from identified MSNs at 14 days in vitro, we are able to detect a much higher rate of inhibitory functional synapses than what has been previously reported in slice recordings. These collateral synapses release γ‐Aminobutyric acid (GABA) and shape the firing patters of other MSNs. Although reduced in vitro models have a number of inherent limitations, the cultures described here provide a unique opportunity to study frequently observed functional collaterals between identifiable MSNs. Additionally, cultured neurons allow for control of the extracellular environment, with the potential to investigate pharmacological regulation of inhibitory MSNs collaterals.