Department of Cell and Molecular Biology, Tulane University, 6400 Freret St, New Orleans, LA 70118, USA.
J Physiol. 2011 Jul 15;589(Pt 14):3517-32. doi: 10.1113/jphysiol.2011.211623. Epub 2011 May 23.
The dentate gyrus of the hippocampus is thought to control information flow into the rest of the hippocampus. Under pathological conditions, such as epilepsy, this protective feature is circumvented and uninhibited activity flows throughout the hippocampus. Many factors can modulate excitability of the dentate gyrus and ultimately, the hippocampus. It is therefore of critical importance to understand the mechanisms involved in regulating excitability in the dentate gyrus. Dynorphin, the endogenous ligand for the kappa (κ) opioid receptor (KOR), is thought to be involved in neuromodulation in the dentate gyrus. Both dynorphin and its receptor are widely expressed in the dentate gyrus and have been implicated in epilepsy and other complex behaviours such as stress-induced deficits in learning and stress-induced depression-like behaviours. Administration of KOR agonists can prevent both the behavioural and electroencephalographic measures of seizures in several different models of epilepsy. Antagonism of the KORs also prevents stress-induced behaviours. This evidence suggests the KORs as possible therapeutic targets for various pathological conditions. In addition, KOR agonists prevent the induction of LTP. Although there are several mechanisms through which dynorphin could mediate these effects, no studies to date investigated the effects of KOR activation on intrinsic membrane properties and cell excitability. We used whole-cell, patch-clamp recordings from acute mouse hippocampus slices to investigate the effect of KOR activation on dentate gyrus granule cell excitability. The agonist U69,593 (U6, 1 μM) resulted in a lower spike threshold, a decreased latency to first spike, an increased spike half-width, and an overall increase in spike number with current injections ranging from 15 to 45 pA. There was also a reduction in the interspike interval (ISI) both early and late in the spike train, with no change in membrane potential or input resistance. Preincubation of the slice with the selective KOR antagonist, nor-binalthorphimine (BNI, 1 μM) inhibited the effect of U6 on the latency to first spike and spike half-width suggesting that these effects are mediated through KORs. The inclusion of GDP-βS (1 mM) in the recording pipette prevented all of the U6 effects, suggesting that all effects are mediated via a G-protein-dependent mechanism. Inclusion of the A-type K+ current blocker, 4-aminopyridine (4-AP, 5 mM) in the pipette also antagonised the effects of U6. Kv4.2 is one of the channel α subunits thought to be responsible for carrying the A-type K+ current. Incubation of hippocampus slices with U6 resulted in a decrease in the Kv4.2 subunit protein at the cell surface. These results are consistent with an increase in cell excitability in response to KOR activation and may reflect new possibilities for additional opioid functions.
海马齿状回被认为控制着信息流入海马体的其余部分。在病理条件下,例如癫痫,这种保护特性被规避,不受抑制的活动会贯穿整个海马体。许多因素可以调节齿状回的兴奋性,最终调节海马体的兴奋性。因此,了解调节齿状回兴奋性的机制至关重要。内源性κ 阿片受体 (KOR) 配体强啡肽 (dynorphin) 被认为参与了齿状回的神经调节。强啡肽及其受体在齿状回中广泛表达,并与癫痫和其他复杂行为(如应激诱导的学习和应激诱导的抑郁样行为缺陷)有关。KOR 激动剂的给药可以预防几种不同癫痫模型中的行为和脑电图测量的癫痫发作。KOR 拮抗剂也可以预防应激诱导的行为。这一证据表明 KOR 可能是各种病理状况的治疗靶点。此外,KOR 激动剂可防止 LTP 的诱导。尽管有几种机制可以介导强啡肽的这些作用,但迄今为止尚无研究调查 KOR 激活对内在膜特性和细胞兴奋性的影响。我们使用急性小鼠海马脑片的全细胞膜片钳记录来研究 KOR 激活对齿状回颗粒细胞兴奋性的影响。激动剂 U69,593(U6,1 μM)导致尖峰阈值降低,首次尖峰潜伏期缩短,尖峰半宽增加,电流注入范围为 15 至 45 pA 时尖峰数量总体增加。在尖峰序列的早期和晚期,尖峰之间的间隔 (ISI) 也减少,而膜电位或输入电阻没有变化。用选择性 KOR 拮抗剂诺比那他明 (BNI,1 μM) 预孵育脑片可抑制 U6 对首次尖峰潜伏期和尖峰半宽的影响,表明这些作用是通过 KOR 介导的。在记录电极中加入 GDP-βS(1 mM)可防止 U6 的所有作用,表明所有作用都是通过 G 蛋白依赖性机制介导的。在记录电极中加入 A 型钾电流阻断剂 4-氨基吡啶 (4-AP,5 mM) 也拮抗了 U6 的作用。Kv4.2 是负责携带 A 型钾电流的通道 α 亚基之一。海马脑片孵育 U6 可导致细胞表面 Kv4.2 亚基蛋白减少。这些结果与 KOR 激活后细胞兴奋性增加一致,可能反映了阿片类药物功能的新可能性。