Ionotropic GABA and glycine receptor subunit composition in human pluripotent stem cell-derived excitatory cortical neurones.
Ionotropic GABA and glycine receptor subunit composition in human pluripotent stem cell-derived excitatory cortical neurones.
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DOI：
10.1113/jphysiol.2014.278994
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发表时间：
2014-10-01
期刊：
影响因子：
--
通讯作者：
Wyllie DJ
中科院分区：
文献类型：
--
作者：
James OT;Livesey MR;Qiu J;Dando O;Bilican B;Haghi G;Rajan R;Burr K;Hardingham GE;Chandran S;Kind PC;Wyllie DJ
我们已经评估，使用全细胞膜片钳记录和RNA测序（RNA-seq），GABAA受体（GABAARs）和士的宁敏感的甘氨酸受体（GlyRs）表达的兴奋性皮质神经元来自人胚胎干细胞（hECN）的属性和组成。激动剂GABA和蝇蕈醇的EC 50值分别为278 μm和182 μm，α2和α3亚基的强RNA-seq信号支持了GABAAR群体显示低激动剂效力的存在。由GABA的EC_（50）浓度诱发的GABAAR介导的电流可被荷包牡丹碱和印防己毒素阻断，IC_（50）值分别为2.7和5.1 μm。通过GABA诱发电流对地西泮的敏感性和对Zn 2+的不敏感性以及加波沙朵的弱直接激动剂作用评估，hECN GABAAR主要含有γ亚基; RNA-seq表明γ2亚基的主要表达。丙泊酚和依托咪酯对GABA诱发电流的增强以及水杨酰肼（SCS）对电流的抑制作用的缺乏表明β2或β3亚基的表达，RNA-seq分析表明hECN GABAAR中β3的强表达。总之，我们的数据支持hECN GABAAR具有α2/3β3γ2亚基组成的观点-该组成在未成熟啮齿动物皮层中也占主导地位。hECN表达的GlyR被甘氨酸激活，EC 50为167 μm。甘氨酸诱发的（500 μm）电流可被士的宁（IC 50 = 630 nm）和印防己毒素（IC 50 = 197 μm）阻断，后者提示存在异聚体受体群体。RNA-seq表明GlyR可能由α2和β亚基组成。
We have assessed, using whole-cell patch-clamp recording and RNA-sequencing (RNA-seq), the properties and composition of GABAA receptors (GABAARs) and strychnine-sensitive glycine receptors (GlyRs) expressed by excitatory cortical neurons derived from human embryonic stem cells (hECNs). The agonists GABA and muscimol gave EC50 values of 278 μm and 182 μm, respectively, and the presence of a GABAAR population displaying low agonist potencies is supported by strong RNA-seq signals for α2 and α3 subunits. GABAAR-mediated currents, evoked by EC50 concentrations of GABA, were blocked by bicuculline and picrotoxin with IC50 values of 2.7 and 5.1 μm, respectively. hECN GABAARs are predominantly γ subunit-containing as assessed by the sensitivity of GABA-evoked currents to diazepam and insensitivity to Zn2+, together with the weak direct agonist action of gaboxadol; RNA-seq indicated a predominant expression of the γ2 subunit. Potentiation of GABA-evoked currents by propofol and etomidate and the lack of inhibition of currents by salicylidine salycylhydrazide (SCS) indicate expression of the β2 or β3 subunit, with RNA-seq analysis indicating strong expression of β3 in hECN GABAARs. Taken together our data support the notion that hECN GABAARs have an α2/3β3γ2 subunit composition – a composition that also predominates in immature rodent cortex. GlyRs expressed by hECNs were activated by glycine with an EC50 of 167 μm. Glycine-evoked (500 μm) currents were blocked by strychnine (IC50 = 630 nm) and picrotoxin (IC50 = 197 μm), where the latter is suggestive of a population of heteromeric receptors. RNA-seq indicates GlyRs are likely to be composed of α2 and β subunits.
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影响因子：
5.3
作者：
Pitt, Samantha J.;Sivilotti, Lucia G.;Beato, Marco
通讯作者：
Beato, Marco
影响因子：
5.5
作者：
Mangin, JA;Baloul, M;Legendre, P
通讯作者：
Legendre, P
影响因子：
16.2
作者：
DRAGUHN, A;VERDORN, TA;SAKMANN, B
通讯作者：
SAKMANN, B
影响因子：
5.5
作者：
Marabelli, Alessandro;Moroni, Mirko;Sivilotti, Lucia G.
通讯作者：
Sivilotti, Lucia G.
DOI：
10.1085/jgp.201010521
发表时间：
2011-02
期刊：
The Journal of general physiology
影响因子：
--
作者：
Krashia P;Lape R;Lodesani F;Colquhoun D;Sivilotti LG
通讯作者：
Sivilotti LG