当前位置: X-MOL 学术Biochem. Moscow Suppl. Ser. A › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Visualization, Properties, and Functions of GABAergic Hippocampal Neurons Containing Calcium-Permeable Kainate and AMPA Receptors
Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Pub Date : 2020-01-01 , DOI: 10.1134/s1990747820010109
V. P. Zinchenko , S. G. Gaidin , I. Yu. Teplov , A. M. Kosenkov , A. I. Sergeev , L. P. Dolgacheva , S. T. Tuleuhanov

Abstract Calcium-permeable kainate (CP-KARs) and AMPA (CP-AMPARs) receptors of the brain neurons are active participants of synaptic plasticity and neurotransmitter release trigger. In this paper, CP-KARs and CP-AMPARs were identified in hippocampal neuroglial culture on 14–17 day of cultivation by a characteristic Ca 2+ response to a selective agonist of CP-KARs and CP-AMPARs, domoic acid (DA), and to a selective agonist of CP-KARs, ATPA. It was shown that DA at a concentration of 300 nM caused a rapid intracellular Ca 2+ concentration increase in two minor subpopulations of neurons. Both subpopulations were found to be GABAergic neurons that were positively stained with antibodies against glutamate decarboxylase 65 and 67 (GAD65/67). The antagonist of CP-AMPARs, NASPM, did not suppress Ca 2+ response to DA in the neurons of the first subpopulation. The selective agonist of CP-KARs, ATPA, increased [Ca 2+ ] i to the same extent as DA only in the first subpopulation of GABAergic neurons. An inhibitor of GABA(A) receptors, bicuculline, did not increase the amplitude of Ca 2+ response to DA in this subpopulation, indicating the absence of CP-KARs in the postsynaptic membrane, where GABA(A) receptors are located. Thus, these GABAergic neurons can be attributed to neurons containing CP-KARs, which are apparently located in the presynaptic membrane of the GABAergic neurons. The [Ca 2+ ] i increase caused by the DA application in the second subpopulation was completely suppressed by NASPM, an inhibitor of CP-AMPARs. NASPM reduced the Ca 2+ oscillations amplitude in the same subset, indicating the involvement of CP-AMPARs in the Ca 2+ impulse formation during synchronous calcium activity. For this reason, the neurons of this subpopulation can be attributed to the GABAergic neurons containing CP-AMPARs. Most of the neurons in the hippocampal cell culture (70–85%) were not stained with antibodies against GAD65/67 and responded to the DA by increasing the calcium oscillations frequency with a delay. The amplitude of DA-induced oscillations increased in the presence of NASPM in the subpopulation of inhibitory neurons containing CP-KARs, indicating their innervation by inhibitory neurons containing CP-AMRARs. This increase in the Ca 2+ oscillation amplitude in the inhibitory neurons containing CP-KARs correlated with a decrease in the amplitude of synchronous calcium activity in a large subpopulation (42 ± 6% of cells) of glutamatergic neurons, suggesting innervation of the latter by inhibitory neurons containing CP-KARs. Thus, GABAergic neurons containing CP-KARs and CP-AMPARs can work in tandem, controlling the activity of individual subpopulations of neurons.

中文翻译:

含有钙渗透性红藻氨酸和 AMPA 受体的 GABA 能海马神经元的可视化、特性和功能

摘要 脑神经元的钙渗透性红藻氨酸(CP-KARs)和AMPA(CP-AMPARs)受体是突触可塑性和神经递质释放触发器的积极参与者。在本文中,通过对 CP-KARs 和 CP-AMPARs 的选择性激动剂软骨藻酸 (DA) 的特征性 Ca 2+ 反应,在培养 14-17 天的海马神经胶质细胞培养物中鉴定出 CP-KARs 和 CP-AMPARs,和 CP-KAR 的选择性激动剂 ATPA。结果表明,浓度为 300 nM 的 DA 在两个较小的神经元亚群中引起细胞内 Ca 2+ 浓度的快速增加。发现这两个亚群都是 GABA 能神经元,它们被针对谷氨酸脱羧酶 65 和 67 (GAD65/67) 的抗体阳性染色。CP-AMPARs 的拮抗剂,NASPM,没有抑制第一亚群神经元中对 DA 的 Ca 2+ 反应。CP-KAR 的选择性激动剂 ATPA 仅在 GABA 能神经元的第一个亚群中将 [Ca 2+ ] i 增加到与 DA 相同的程度。GABA(A) 受体的抑制剂荷包牡丹碱不会增加该亚群中对 DA 的 Ca 2+ 反应的幅度,表明在 GABA(A) 受体所在的突触后膜中不存在 CP-KAR。因此,这些 GABA 能神经元可归因于含有 CP-KAR 的神经元,它们显然位于 GABA 能神经元的突触前膜中。由 DA 应用在第二个亚群中引起的 [Ca 2+ ] i 增加被 NASPM(CP-AMPAR 的抑制剂)完全抑制。NASPM 降低了同一子集中的 Ca 2+ 振荡幅度,表明 CP-AMPAR 在同步钙活动期间参与 Ca 2+ 脉冲形成。因此,该亚群的神经元可归因于含有 CP-AMPAR 的 GABA 能神经元。海马细胞培养物中的大多数神经元(70-85%)没有被针对 GAD65/67 的抗体染色,并且通过延迟增加钙振荡频率来响应 DA。在含有 CP-KARs 的抑制性神经元亚群中存在 NASPM 时,DA 诱导的振荡幅度增加,表明它们受到含有 CP-AMRARs 的抑制性神经元的支配。含有 CP-KARs 的抑制性神经元中 Ca 2+ 振荡幅度的增加与谷氨酸能神经元的大亚群(细胞的 42 ± 6%)中同步钙活性幅度的降低相关,表明后者的神经支配含有 CP-KAR 的抑制性神经元。因此,含有 CP-KAR 和 CP-AMPAR 的 GABA 能神经元可以协同工作,控制单个神经元亚群的活动。
更新日期:2020-01-01
down
wechat
bug