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Synaptic Plasticity

開催日 2014/9/11
時間 16:00 - 17:00
会場 Poster / Exhibition(Event Hall B)

Involvement of N-glycosylation in AMPA receptor channel properties

  • P1-052
  • 若園 佳彦 / Yoshihiko Wakazono:1 Kandel Munal B. / Munal B. Kandel:1 緑川 良介 / Ryosuke Midorikawa:1 川崎 ナナ / Nana Kawasaki:2 岡 昌吾 / Shogo Oka:3 髙宮 考悟 / Kogo Takamiya:1 
  • 1:宮崎大・医・統合生理 / Dept Integrative Physiol, Faculty of Med, Univ of Miyazaki, Miyazaki, Japan 2:国立医薬品食品衛生研・生物薬品部 / Div Biol Chem and Biologicals, Natl Inst Health Sci, Japan 3:京都大院・医・人間健康科学・生化 / Dept Biol Chem, Human Health Sci, School of Med, Kyoto Univ, Kyoto, Japan 

The intracellular molecular mechanisms underlying the regulation of the AMPA receptor have been dramatically elucidated in the past few decades. In contrast, the regulation of the extracellular domain remains unclear. Here, we focused on N-glycosylation of the AMPA receptor in the extracellular domain and tried to clarify their functions by combining molecular biological and electrophysiological techniques.
First, we examined the effects of PNGase F, which is a glycosidase that cleaves the N-glycosylation site at an asparagine residue, on AMPA currents in primary hippocampal cultured neurons and HEK293 cells expressing GluA1 using a whole-cell patch-clamp technique. Glutamate response elicited with puff application demonstrated that the digestion of N-glycosylation induced the re-sensitization of AMPA currents in the both. It is reported that six putative N-glycosylation sites are located in the extracellular domain of GluA1. Next, we performed mutation studies of each N-glycosylation site of GluA1 to identify the N-glycosylation site responsible for the re-sensitization induced by PNGase F treatment. These results indicated that N401 was a critical site for the expression of re-sensitization. Finally, we analyzed the possibility of re-sensitization after PNGase F treatment using acute brain slices. Single electrical stimulation of Schaffer collateral did not show the re-sensitization in hippocampal pyramidal neurons, however, paired pulse stimulation generated the re-sensitization.
Taken together, these results suggest that N-glycosylation, especially at N401 site, can modulate AMPA receptor channel properties.

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