• Title/Summary/Keyword: Postsynaptic density

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Nonspecific Association of a 17 kDa Isoform of the Myelin Basic Protein with the Postsynaptic Density Fraction

  • Moon, Il-Soo
    • BMB Reports
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    • v.33 no.3
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    • pp.276-278
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    • 2000
  • The postsynaptic density (PSD), a large protein complex beneath the postsynaptic membrane, is notorious for its 'stickiness'. In order to understand the molecular composition of the PSD fraction, a 17 kDa protein band was isolated by electroelution from SDS-geis, and its partial amino acid sequence was determined from HPLC-purified tryptic peptides of the protein. Surprisingly, the amino acid sequence was identical to that of the previously reported 17 kDa isoform of the myelin basic protein (MBP), an essential protein in CNS myelin formation. Since the protein band represented ~2% of the total proteins in the 1 % n-octyl glucoside-insoluble PSD fraction, these results indicate that a significant amount of the 17 kDa isoform of MBP is tightly associated with the PSD during preparation of the PSD fraction.

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Proteins in the Postsynaptic Density of the Central Nervous System

  • Moon, Il-Soo
    • Journal of Life Science
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    • v.9 no.2
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    • pp.34-39
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    • 1999
  • The postsynaptic density (PSD) is a cytoskeletal specialization that is involved in the regulation of synaptic signal transduction. Mainly due to the hydrophobic nature of the PSD proteins, characterization of this intriguing structure at the molecular level has been very intractable until early 1990s. However, recent development in protein microchemistry and molecular cloning techniques allowed identification and characterization of the PSD proteins. As expected, cytoskeletal proteins constitute major components of the PSD. Other major PSD proteins have been identified by protein sequencing, and their genes were used to fish out associating proteins by yeast two-hybrid system expanding our knowledge on the molecular structure of the PSD significantly. In this review, I summarize proteins that are so far identified focusing on the glutamatergic synapses.

Electron Tomography and Synapse Study

  • Kim, Hyun-Wook;Kim, Dasom;Rhyu, Im Joo
    • Applied Microscopy
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    • v.44 no.3
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    • pp.83-87
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    • 2014
  • Electron tomography (ET) is a useful tool to investigate three-dimensional details based on virtual slices of relative thick specimen, and it requires complicated procedures consisted of image acquisition steps and image processing steps with computer program. Although the complicated step, this technique allows us to overcome some limitations of conventional transmission electron microscopy: (1) overlapping of information in the ultrathin section covering from 30 nm to 90 nm when we observe very small structures, (2) fragmentation of the information when we study larger structures over 100 nm. There are remarkable biological findings with ET, especially in the field of neuroscience, although it is not popular yet. Understanding of behavior of synaptic vesicle, active zone, pooling and fusion in the presynaptic terminal have been enhanced thanks to ET. Some sophisticated models of postsynaptic density with ET and immune labeling are introduced recently. In this review, we introduce principles, practical steps of ET and some recent researches in synapse biology.

Oxygen/Glucose Deprivation and Reperfusion Cause Modifications of Postsynaptic Morphology and Activity in the CA3 Area of Organotypic Hippocampal Slice Cultures

  • Jung, Yeon Joo;Suh, Eun Cheng;Lee, Kyung Eun
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.6
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    • pp.423-429
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    • 2012
  • Brain ischemia leads to overstimulation of N-methyl-D-aspartate (NMDA) receptors, referred as excitotoxicity, which mediates neuronal cell death. However, less attention has been paid to changes in synaptic activity and morphology that could have an important impact on cell function and survival following ischemic insult. In this study, we investigated the effects of reperfusion after oxygen/glucose deprivation (OGD) not only upon neuronal cell death, but also on ultrastructural and biochemical characteristics of postsynaptic density (PSD) protein, in the stratum lucidum of the CA3 area in organotypic hippocampal slice cultures. After OGD/reperfusion, neurons were found to be damaged; the organelles such as mitochondria, endoplasmic reticulum, dendrites, and synaptic terminals were swollen; and the PSD became thicker and irregular. Ethanolic phosphotungstic acid staining showed that the density of PSD was significantly decreased, and the thickness and length of the PSD were significantly increased in the OGD/reperfusion group compared to the control. The levels of PSD proteins, including PSD-95, NMDA receptor 1, NMDA receptor 2B, and calcium/calmodulin-dependent protein kinase II, were significantly decreased following OGD/reperfusion. These results suggest that OGD/reperfusion induces significant modifications to PSDs in the CA3 area of organotypic hippocampal slice cultures, both morphologically and biochemically, and this may contribute to neuronal cell death and synaptic dysfunction after OGD/reperfusion.

Studies on the phosphotyrsine-proteins in the rat cerbellar PSD fraction (흰쥐 소뇌 연접후치밀질내 phosphotryrosine 함유 단백질에 대한 연구)

  • 전일수;함소희;고복현
    • Journal of Life Science
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    • v.7 no.3
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    • pp.198-204
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    • 1997
  • The signal transduction through tyrosine kinases play important roles in neuronal development and synaptic regulation. We carried out immunoblot analyses to study tyrosine=phosphorylated proteins in the rat cerebellar postsynaptic density (PSD), a protein-rich cytoskeletal specialization underlying beneath the postsynaptic membrane. The overall protein composition of cerebellar PSD fractions was similar to that of the forebrain’s and only a few bands were different in Coomassie stain. Immunoblot analyses with phosphtyrosine-specific antiboy (4G10) showed that there are many more tyrosine-phosphorylated proteins in the cerebellar PSD than in the forebrain PSD. Interestiingly, a major phosphotyrosine signals in cerebellar PSD fractions was associated with a 50 kD molecular size, named as PSD-50. Migration of PSD-50 coincided with that of $\alpha$CaMKII and remained in the pellet fraction after N-octylglucoside extraction. These results indicate that tyrosine phosphorylation is important in cerebellar synaptic regulation and that the PSD-50 may be same as $\alpha$CaMKIIor a new protein which is a major substrate of tyrosine kinase.

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Localization of Translation Initiation Factors to the Postsynaptic Sites (신경세포 연접후 위치에 단백질합성 해석시작인자(eIF)들의 존재)

  • Choi, Myoung-Kwon;Park, Sung-Dong;Park, In-Sick;Moon, Il-Soo
    • Journal of Life Science
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    • v.21 no.11
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    • pp.1526-1531
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    • 2011
  • Local protein synthesis in neuronal dendrites is important for site-specific regulation of synaptic plasticity. In this study, we investigated whether translation initiation factors (eIFs) are present at the postsynaptic sites. High resolution confocal microscopy showed that the eIF4E and eIF4G (which bind the 5'-terminal mRNA cap), eIF5 (which is important during the 3' direction scanning to find an initiation codon), eIF6 (which mediates upregulation of translation by external stimuli), and eIF5A (which mediate translation upregulation under adverse conditions) were localized to the post-synaptic sites. Immunoblot and detergent extraction experiments also indicated that these eIFs were present in the synapse in association with the postsynaptic density (PSD). Our data provide evidence for the strategic positioning of eIFs at the postsynaptic site for initiation of translation in diverse situations.

Ultrastructural Analysis of Chemical Synapses in Cultured Wild Type Drosophila Embryonic Neurons (초파리 배자 신경세포의 화학적 신경연접 미세구조)

  • Oh, Hyun-Woo;Park, Ho-Yong
    • Applied Microscopy
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    • v.34 no.4
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    • pp.223-230
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    • 2004
  • To identify the structural basis of mutations that affect synaptic transmission we have begun quantitative ultrastructural descriptions of synapses in cultured Drosophila embryonic neurons. In wild-type cultures, synapses are distinguished by the parallel arrangement of a thickened pre- and post synaptic membrane separated by a synaptic cleft. The presynaptic active zones and postsynaptic densities are defined by electron dense material close to the membrane. Presynaptic regions are also characterized by the presence of one or more electron dense regions, presynaptic densities, around which a variable number of small, clear core synaptic vesicles (mean $35.1{\pm}1.44$ nm in diameter) are clustered. Subsets of these vesicles are in direct contact with either the presynaptic density or the membrane and are considered morphologically docked. A small number of larger, dense core vesicles are also observed in most presynaptic profiles.

An efficient and reliable electroelution method from SDS-PAGE: Identification of a 31 kDa protein in the postsynaptic density fraction as adenine nucleotide translocator 1 (SDS-겔로부터 효율적인 단백질 분리방법과 31 kDa 연접후치밀질 단백질의 동정)

  • Jung, Jae-Seob;Cho, Sun-Jung;Shin, Seung-Chul;Jin, Ing-Nyol;Jung, Yong-Wook;Ko, Bok-Hyun;Moon, Il-Soo
    • Journal of Life Science
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    • v.12 no.5
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    • pp.555-560
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    • 2002
  • The molecular composition of the postsynaptic density (PSD) is largely hon. In this report, an electroelution protocol was demonstrated to be used for efficient isolation of PSD proteins with diverse molecular sizes. Using this protocol, a 31 kDa protein in the 1% n-octyl glucoside-insoluble PSD fraction (termed as PSD31) was purified from SDS-gels, and internal peptides were determined for amino acid sequences. The amino acid sequences of the PSD31 were highly homologous with the adenine nucleotide translocator 1 (ANTI). The association of ANTl with PSD suggests presence of a mechanism in synapses for releasing adenosine nucleotides into the extracellular space.

The Memory-Enhancing Effects of Liquiritigenin by Activation of NMDA Receptors and the CREB Signaling Pathway in Mice

  • Ko, Yong-Hyun;Kwon, Seung-Hwan;Hwang, Ji-Young;Kim, Kyung-In;Seo, Jee-Yeon;Nguyen, Thi-Lien;Lee, Seok-Yong;Kim, Hyoung-Chun;Jang, Choon-Gon
    • Biomolecules & Therapeutics
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    • v.26 no.2
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    • pp.109-114
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    • 2018
  • Liquiritigenin (LQ) is a flavonoid that can be isolated from Glycyrrhiza radix. It is frequently used as a tranditional oriental medicine herbal treatment for swelling and injury and for detoxification. However, the effects of LQ on cognitive function have not been fully explored. In this study, we evaluated the memory-enhancing effects of LQ and the underlying mechanisms with a focus on the N-methyl-D-aspartic acid receptor (NMDAR) in mice. Learning and memory ability were evaluated with the Y-maze and passive avoidance tests following administration of LQ. In addition, the expression of NMDAR subunits 1, 2A, and 2B; postsynaptic density-95 (PSD-95); phosphorylation of $Ca^{2+}$/calmodulin-dependent protein kinase II (CaMKII); phosphorylation of extracellular signal-regulated kinase 1/2 (ERK 1/2); and phosphorylation of cAMP response element binding (CREB) proteins were examined by Western blot. In vivo, we found that treatment with LQ significantly improved memory performance in both behavioral tests. In vitro, LQ significantly increased NMDARs in the hippocampus. Furthermore, LQ significantly increased PSD-95 expression as well as CaMKII, ERK, and CREB phosphorylation in the hippocampus. Taken together, our results suggest that LQ has cognition enhancing activities and that these effects are mediated, in part, by activation of the NMDAR and CREB signaling pathways.

Effects of Chronic Treatment of Taegeuk Ginseng on Cognitive Function Improvement in Scopolamine Induced Memory Retarded Rats (태극삼의 장기투여가 인지기능향상과 기억력증진에 미치는 영향)

  • Lee, Cheol-Hyeong;Park, Ji Hye;Kim, Kyu Il;Lee, Seoul
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.36 no.1
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    • pp.18-22
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    • 2022
  • To investigate effects of cognitive function improvement whether against Taegeuk ginseng on scopolamine-induced memory impairment in rats. All experiments were conducted in three groups: the control group (CTR), the scopolamine 0.4mg/kg (SCP), and the scopolamine (SCP+T) treated with Taegeuk ginseng 100 mg/kg. Taegeuk ginseng 100 mg/kg daily was orally administered for one month and treated with scopolamine was only for 7 consecutive days on the Morris water maze task. 3 weeks after oral administration of Taegeuk ginseng, subjects were performed the Morris water maze test for 8 days and then the open-field exploration test which to assessed for cognitive function improvement. After behavioral testing, subjects were sacrificed and microdissected brains for neurochemical analysis. In the cognitive-behavioral test, long-term administration of Taegeuk ginseng improved spatial navigation learning task compared with the impeded by scopolamine treatment. In neurochemistry, the expression of the synaptic marker PSD95 (postsynaptic density protein 95) was increased in the hippocampus compared to the scopolamine group. Also, brain-derived neurotrophic factor (BDNF) expression was significantly increased in the taegeuk ginseng administration group. These data suggested that long-term administration of taegeuk ginseng might improve cognitive-behavioral functions on hippocampal related spatial learning memory, and it was correlated with neurotropic and synaptic reinforcement. In conclusion, treatment with taegeuk ginseng may positive outcome on learning and memory deficit disorders.