Piochon Claire, Irinopoulou Theano, Brusciano Daniel, Bailly Yannick, Mariani Jean, Levenes Carole
Université Pierre et Marie Curie-Paris 6, Unité Mixte de Recherche 7102, Centre National de la Recherche Scientifique, Laboratoire Neurobiologie des Processus Adaptifs, Equipe Développement et Vieillissement du Système Nerveux, 75005 Paris, France.
J Neurosci. 2007 Oct 3;27(40):10797-809. doi: 10.1523/JNEUROSCI.2422-07.2007.
Among integrative neurons displaying long-term synaptic plasticity, adult Purkinje cells seemed to be an exception by lacking functional NMDA receptors (NMDA-Rs). Although numerous anatomical studies have shown both NR1 and NR2 NMDA-R subunits in adult Purkinje cells, patch-clamp studies failed to detect any NMDA currents. Using more recent pharmacological and immunodetection tools, we demonstrate here that Purkinje cells from adult mice respond to exogenous NMDA application and that postsynaptic NMDA-Rs carry part of the climbing fiber-mediated EPSC (CF-EPSC), with undetectable contribution from presynaptic or polysynaptic NMDA currents. We also detect NR2-A/B subunits in adult Purkinje cells by immunohistochemistry. The NMDA-mediated CF-EPSC is barely detectable before 3 weeks postnatal. From the end of the third week, the number of cells displaying the NMDA-mediated CF-EPSC rapidly increases. Soon, this EPSC becomes detectable in all the Purkinje cells but is still very small. Its amplitude continues to increase until 12 weeks after birth. In mature Purkinje cells, we show that the NMDA-Rs contribute to the depolarizing plateau of complex spikes and increase their number of spikelets. Together, these observations demonstrate that mature Purkinje cells express functional NMDA receptors that become detectable in CF-EPSCs at approximately 21 d after birth and control the complex spike waveform.
在表现出长期突触可塑性的整合神经元中,成年浦肯野细胞似乎是个例外,因为它们缺乏功能性N-甲基-D-天冬氨酸受体(NMDA-Rs)。尽管众多解剖学研究已在成年浦肯野细胞中发现了NR1和NR2 NMDA-R亚基,但膜片钳研究未能检测到任何NMDA电流。利用更新的药理学和免疫检测工具,我们在此证明,成年小鼠的浦肯野细胞对外源性NMDA的应用有反应,并且突触后NMDA-Rs介导了部分攀爬纤维介导的兴奋性突触后电流(CF-EPSC),而突触前或多突触NMDA电流的贡献无法检测到。我们还通过免疫组织化学在成年浦肯野细胞中检测到了NR2-A/B亚基。NMDA介导的CF-EPSC在出生后3周之前几乎检测不到。从第三周结束时起,显示NMDA介导的CF-EPSC的细胞数量迅速增加。很快,这种EPSC在所有浦肯野细胞中都可检测到,但仍然非常小。其幅度持续增加,直到出生后12周。在成熟的浦肯野细胞中,我们表明NMDA-Rs有助于复合动作电位的去极化平台期并增加其棘波数量。总之,这些观察结果表明,成熟的浦肯野细胞表达功能性NMDA受体,这些受体在出生后约21天时在CF-EPSCs中变得可检测到,并控制复合动作电位波形。