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Protrusion of N-acetylglucosamine Kinase Clusters into the Base of Excitatory Synapses

흥분성연접의 바닥으로 NAGK 클러스터의 돌출

  • Moon, Il-Soo (Department of Anatomy, College of Medicine, Dongguk University) ;
  • Cho, Sun-Jung (Department of Anatomy, College of Medicine, Dongguk University) ;
  • Lee, Hyun-Sook (Department of Anatomy, College of Medicine, Dongguk University) ;
  • Seog, Dae-Hyun (Department of Biochemistry, College of Medicine, Inje University) ;
  • Walikonis, Randall (Department of Physiology and Neurobiology, University of Connecticut)
  • Published : 2009.08.30

Abstract

N-Acetylglucosamine kinase (GlcNAc kinase or NAGK; EC 2.7.1.59) catalyzes the phosphorylation of GlcNAc to GlcNAc-6-phosphate (GlcNAc-6-P). Despite detailed characterization of the enzyme itself, there have been few studies on the expression of NAGK in mammalian tissues. In the rat hippocampal neuron in culture, NAGK-immunoreactivity (IR) formed clusters in somatodendritic domains. In this study we characterized the NAGK clusters that protrude out the long axis of dendritic shafts. By double-labeling of the neurons with antibodies against NAGK and various synaptic proteins, we show that NAGK is positioned at the base of spines, while there were no NAGK protrusions into inhibitory postsynaptic sites. Immunoblot analysis showed that NAGK was included in synaptosomes but not in PSD fractions. Our results indicate that the NAGK clusters at the dendritic periphery protrude into spines.

N-Acetylglucosamine kinase (GlcNAc kinase or NAGK; EC 2.7.1.59)는 GlcNAc를 인산화하여 GlcNAc-6-phosphate를 만드는 효소이다. 효소자체에 대한 자세한 연구에도 불구하고 포유류에서NAGK의 표현에 대한 연구는 거의 없다. 배양한 흰쥐의 해마신경세포에서 NAGK은 세포체/가지돌기 영역에서 클러스터(cluster)를 형성한다. 본 연구에서는 가지돌기의 긴 축에서부터 가쪽으로 돌출되는 NAGK 클러스터에 대하여 연구하였다. 배양한 해마신경세포를 NAGK와 다양한 연접표지단백질에 대한 항체로 이중염색한 결과 NAGK 클러스터가 가지돌기의 바닥에는 있었지만 억제성 연접후부위에는 존재하지 않았다. 또한 흰쥐 전뇌(forebrain)의 균질액(homogenate, BH), 연접체(synaptosome, S), 연접후치밀질(postsynaptic density, PSD) 분획을 NAGK 항체로 면역염색한 결과 NAGK는 연접체에는 있었지만 PSD 분획에는 존재하지 않았다. 이러한 결과들은 NAGK가 가지돌기가시(spine)의 바닥쪽으로 돌출됨을 의미한다.

Keywords

References

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