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发育中小鼠海马神经元中的突触N-甲基-D-天冬氨酸受体:功能特性及对艾芬地尔的敏感性

Synaptic NMDA receptors in developing mouse hippocampal neurones: functional properties and sensitivity to ifenprodil.

作者信息

Kirson E D, Yaari Y

机构信息

Department of Physiology, Hebrew University-Hadassah School of Medicine, Jerusalem, Israel.

出版信息

J Physiol. 1996 Dec 1;497 ( Pt 2)(Pt 2):437-55. doi: 10.1113/jphysiol.1996.sp021779.

Abstract
  1. Whole-cell patch-clamp techniques were used to record pharmacologically isolated NMDA receptor-mediated EPSCs (NMDA EPSCs) from CA1 pyramidal cells (PCs) in hippocampal slices from 4-day-old to 36-week-old mice, in order to characterize developmental changes in functional properties and subunit composition of synaptic NMDA receptors. 2. During the first postnatal weeks the dendritic tree of CA1 PCs stained with biocytin increased both in size and in complexity. This was associated with an increase in amplitude of the focally evoked NMDA EPSCs recorded either in nominally Mg(2+)-free or Mg(2+)-containing saline. In adult PCs (> 5 weeks old) EPSC amplitude was 4-fold larger than in very young (up to 2 weeks old) neurones. 3. The sensitivity of NMDA EPSCs to blockade by Mg2+ did not change with age. In very young, intermediate and adult PCs the EPSC-voltage relation displayed an area of negative slope conductance at membrane potentials more negative than -30 mV. The apparent Kd values of the NMDA receptors for Mg2+ at 0 mV were 7.8 +/- 6.4, 10.4 +/- 14.1 and 6.5 +/- 4.7 mM in very young, intermediate and adult neurones, respectively. 4. The decay of the NMDA EPSC in both young and adult neurones could be described by the sum of a fast and a slow exponential function. Both EPSC rise time and fast and slow decay time constants measured at -60 mV, decreased with age. 5. The decay of NMDA EPSCs in young versus adult PCs was differentially modulated by membrane voltage. In young PCs depolarization slowed both the fast and the slow EPSC components. In adult PCs depolarization slightly accelerated the initial EPSC decay, though the overall duration of the EPSC did not change. The rise time of the EPSCs was not affected by voltage at any age. 6. The subunit-selective NMDA receptor antagonist ifenprodil similarly blocked iontophoretic NMDA-induced currents and NMDA EPSCs. In both young and adult PCs, the concentration-response curves for this effect disclosed distinct low and high affinity binding sites for ifenprodil. 7. In young PCs, low and high affinity binding sites for ifenprodil were about equally expressed (57 versus 43%, respectively), whereas in adult PCs, synaptic NMDA receptors expressed a majority (78%) of low affinity binding sites for ifenprodil. 8. The long duration of NMDA EPSCs (and by implication, of Ca2+ transfer through NMDA receptor channels) and its further prolongation by depolarization in young PCs are consistent with heightened NMDA-dependent neuronal plasticity early in development. The age-related changes in these properties may result from a developmental change in NMDA receptor subunit composition.
摘要
  1. 采用全细胞膜片钳技术记录4日龄至36周龄小鼠海马切片中CA1锥体神经元(PCs)上药理学分离的N-甲基-D-天冬氨酸受体介导的兴奋性突触后电流(NMDA EPSCs),以表征突触NMDA受体功能特性和亚基组成的发育变化。2. 在出生后的最初几周,用生物素染色的CA1 PCs的树突树在大小和复杂性上均增加。这与在无镁或含镁盐水中记录到的局部诱发NMDA EPSCs的幅度增加有关。在成年PCs(>5周龄)中,EPSC幅度比非常年幼(至多2周龄)的神经元大4倍。3. NMDA EPSCs对镁离子阻断的敏感性不随年龄变化。在非常年幼、中等年龄和成年PCs中,EPSC-电压关系在膜电位比-30 mV更负时显示出负斜率电导区域。在0 mV时,非常年幼、中等年龄和成年神经元中NMDA受体对镁离子的表观解离常数(Kd)值分别为7.8±6.4、10.4±14.1和6.5±4.7 mM。4. 年轻和成年神经元中NMDA EPSC的衰减都可以用一个快速指数函数和一个慢速指数函数之和来描述。在-60 mV测量的EPSC上升时间以及快速和慢速衰减时间常数均随年龄减小。5. 年轻与成年PCs中NMDA EPSCs的衰减受到膜电压的不同调节。在年轻PCs中,去极化减慢了快速和慢速EPSC成分。在成年PCs中,去极化略微加速了初始EPSC衰减,尽管EPSC的总持续时间没有变化。EPSCs的上升时间在任何年龄都不受电压影响。6. 亚基选择性NMDA受体拮抗剂ifenprodil同样阻断离子电泳NMDA诱导的电流和NMDA EPSCs。在年轻和成年PCs中,这种效应的浓度-反应曲线揭示了ifenprodil明显的低亲和力和高亲和力结合位点。7. 在年轻PCs中,ifenprodil的低亲和力和高亲和力结合位点表达大致相等(分别为57%和43%),而在成年PCs中,突触NMDA受体表达了大部分(78%)ifenprodil的低亲和力结合位点。8. NMDA EPSCs的长时间持续(以及由此暗示的通过NMDA受体通道的钙离子转运)及其在年轻PCs中通过去极化的进一步延长与发育早期NMDA依赖性神经元可塑性增强一致。这些特性中与年龄相关的变化可能是由于NMDA受体亚基组成的发育变化所致。
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e91e/1160995/f3aacb052b50/jphysiol00386-0141-a.jpg

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