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3-Dimensional Reconstruction of Parallel fiber-Purkinje Cell Synapses Using High-Voltage Electron Microscopy (고압전자현미경을 이용한 소뇌 평행섬유-조롱박세포간 신경연접의 3차원 재구성)

  • Lee, Kea-Joo;Kweon, Hee-Seok;Kang, Ji-Seoun;Rhyu, Im-Joo
    • Applied Microscopy
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    • v.35 no.1
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    • pp.31-39
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    • 2005
  • Synapses are contact points where one neuron communicates with another. The morphological change of synapses under various physiological or pathological conditions has long been hypothesized to modify their functional properties. 3-dimensional (3-D) reconstruction of synapses with serial ultrathin sections has contributed to the understanding of ultrastructural dimensions and compositions of synapses. The 3-D reconstruction procedures, however, require a great amount of expertise as well as include prohibitively timeconsuming processes. Here, we introduce efficient 3-D reconstruction technique using high-voltage electron microscopy (HVEM). Primarily, we established an optimal section thickness and staining condition to observe synaptic structures in detail under HVEM. The result showed that synaptic profiles were preserved at the section thickness of 250 nm without the overlapping of synaptic ultrastructures. An increase in the reaction time of en bloc staining was most efficient to enhance contrast than the extension of postembedding staining or the addition of uranyl acetate during dehydration. Then, 3-D reconstruction of parallel fiber-Purkinje cell synapses in the rat cerebellum was carried out with serial HVEM images and reconstruction software. The images were aligned and the contours of synapses were outlined on each section. 3-D synapses were finally extracted from the section files by grouping all the synaptic contours. The reconstructed synapse model clearly demonstrated the configuration of pre and postsynaptic components. These results suggest that 3-D reconstruction of synapses using HVEM is much efficient and suitable for massive quantitative studies on synaptic connectivity than conventional TEM approach using numerous ultrathin sections.

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.

Performance Analysis of MAP Algorithm and Concatenated Codes Using Trellis of Block Codes (블록부호의 트렐리스를 이용한 MAP 알고리즘 및 연접부호의 성능분석)

  • 백동철;양경철
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.24 no.6A
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    • pp.905-912
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    • 1999
  • In this paper we explain a trellis representation of block codes and derive their MAP decoding algorithm based on it. We also analyze the performance of block codes and concatenated codes with block codes as components by computer simulations, which were performed by changing the structures and constituent codes of concatenated codes. Computer simulations show that soft decision decoding of block codes get an extra coding gain than their hard decision decoding and that concatenated codes using block codes have good performance in the case of high code rate.

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Ultrastructural Observations of Glutamatergic Synaptic Components in the Basilar Pontine Nuclei of the Dog (개의 교핵내 glutamate성 연접 성분의 미세구조적 위치관찰)

  • Lee, Hyun-Sook
    • Applied Microscopy
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    • v.27 no.1
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    • pp.57-70
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    • 1997
  • The distribution of glutamatergic synaptic structures in the dog basilar pons was investigated at the ultrastructural level using monoclonal antibodies against fixative-modified glutamate. Electron-dense reaction product was densely localized at the perinuclear region in the neurenal somata and often observed along the microtubules located within the dendritic processes. One or more unlabelled axon terminals made asymmetric synaptic contacts with glutamate-immunoreactive dendritic profiles. In audition, reaction product was observed either within axonal processes surrounded by myelin sheath or axon terminals. Immunoreactive axon terminals made asymmetric synaptic contact either with unlabelled or labelled dendritic profiles. These observations provided an anatomic evidence of how this excitatory neural element might perform its function in a multisynaptic pathway involving glutamatergic afferents to the basilar pons, glutamate-immunoreactive pontocerebellar projection neurons, and the glutamate-positive granule cells of the cerebellar cortex.

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Ultrastructural Localization of GABAergic Neuronal Components in the Dog Basilar Pons (개의 교핵내 GABA성 신경세포 성분의 미세구조적 위치관찰)

  • Lee, Hyun-Sook
    • Applied Microscopy
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    • v.25 no.1
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    • pp.65-74
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    • 1995
  • An immunocytochemical study of GABA-positive neuronal elements was performed at the electron microscopic level to examine subcellular distribution of the inhibitory neurotransmitter in the dog basilar pons. Electron-dense reaction product was observed in neuronal somata and dendritic processes. One or more unlabeled axon terminals made asymmetric synaptic contacts with these GABAergic somatic and dendritic profiles. A large number of GABA-positive axon terminals were also observed. They made symmetric as well as asymmetric synaptic contacts with unlabeled dendritic profiles. In axo-axonic synapses, postsynaptic axon-like processes were consistently GABA-immunoreactive. These observations suggest that the inhibitory local circuit neurons in the dog basilar pons play a major role in cerebro-ponto-cerebellar circuitry by integrating various afferent inputs and conveying them into the cerebellar cortex and the deep cerebellar nuclei.

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MORPHOLOGY OF THE TERMINAL ARBORS FROM THE MASSETERIC MUSCLE SPINDLE AFFERENTS IN THE TRIGEMINAL MOTOR NUCLEUS (삼차신경 운동핵에서 교근 근방추 구심성 신경섬유 종말지의 미세구조)

  • Lee, Kyung-Woo;Bae, Yong-Chul;Kim, Chin-Soo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.16 no.3
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    • pp.321-347
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    • 1994
  • Muscle spindle afferents from masseter muscle were labelled by the intra-axonal HRP injection and were processed for light microscopic reconstruction. Regions containing terminal arbors scattered in the central portion of the masseteric motor neuron pool (type I a) and those restricted to 2-3 small portion of it (type II) were selected and processed for electronmicroscopic analysis with serial sections. The shape of the labelled boutons was dome or elongated shape. Scalloped or glomerulus shape with peripherial indentation containing pre or postsynaptic neuronal propiles, which is occasionally found in the trigeminal main sensory nucleus and spinal dorsal horn, was not observed. Both type Ia and type II boutons had pale axoplasm and contained clear, spherical vesicles of uniform size(dia : 49-52nm) and occasionally large dense cored vesicles(dia : 87-118nm). The synaptic vesicles were evenly distributed throughout the boutons although there was a slight tendency of vesicles to accumulate at the presynaptic site. The average of short and long diameter(short D. + long D./2) of type I a bouton was smaller than that of type II bouton. All the labelled boutons, which showed prominent postsynaptic density, large synaptic area and multiple synaptic contact, made asymmetrical synaptic contact with postsynaptic neuronal propiles. Most of the type Ia and type II boutons made synaptic contact with only one neuronal propile and boutons which shows synaptic contact or more neuronal propiles was not observed. Most of the type Ia boutons(87.2%) were presynaptic to the soma or proximal dendrite and a few remainder(12.8%) made synaptic contact with dendritic shaft or distal dendrite. In contrast, majority of type II boutons showed synaptic contact with dendritic shaft and remainder with soma or proximal dendrite. In conclusion, terminal boutons which participate in the excitatory monosynaptic jaw jerk reflex made synaptic contact with more proximal region of the neuron, and showed very simple synaptic connection, compared with those from the primary afferenst in the other region of the central nervous system such as spinal dorsal horn and trigeminal main sensory nucleus which assumed to be responsible for the mediating pain, tactile sensation, sensory processing or sensory discrimination.

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ULTRASTRUCTURAL ANALYSIS OF TOOTH PULP AFFERENTS TERMINALS IN THE MEDULLARY DORSAL HORN OF THE RAT (치수유래 구심성 신경섬유의 삼차신경 감각핵군에서의 연접특성)

  • Bae, Yong-Chul;Lee, Eun-Hee;Choy, Min-Ki;Hong, Su-Hyung;Kim, Hyun-Jung;Na, Soon-Hyeun;Kim, Young-Jin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.2
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    • pp.219-227
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    • 2001
  • Little is known about processing mechanism of pain sensation of the oral cavity at the 1st synapse of trigeminal sensory nuclei. Serial ultrathin sections of tooth pulp afferent terminals, identified by the transganglionic transport of 1% wheatgerm agglutinin conjugated horseradish peroxidase, were investigated with electron microscope. Quantitative ultrastructural analysis was performed on digitizing tablet connected to Macintoshi personal computer (software; NIH Image 1.60, NIH, Bethesda, MD). Labeled boutons could be classified into two types by the shapes of containing vesicles : S bouton, which contained mainly spherical vesicles (Dia. 45-55 nm) and few large dense cored vesicles (Dia, 80-120nm), and LDCV bouton, which contained spherical vesicles as well as large number of large dense cored vesicles. Most of the parameters on the ultrastructural characteristic and synaptic organization of labeled boutons were similar between S and LDCV boutons, except shapes of containing vesicles. Majority of the labeled boutons showed simple synaptic arrangement. The labeled boutons were frequency presynaptic to dendritic spine, and to a lesser extent, dendritic shaft. They rarely synapsed with soma and adjacent proximal dendrite. A small proportion of labeled boutons made synaptic contacts with presynaptic, pleomorphic vesicles containing endings and synaptic triad. Morphometric parameters of labeled boutons including volume and surface area, total apposed area, mitochondrial volume, active zone area, vesicle number and density showed wide variation and these were not significantly different between S and LDCV boutons. The present study revealed characteristic features on ultrastructure and synaptic connection of pulpal afferents which may involved in transmission of oral pain sensation.

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Cerebral cortical or Cerebellar Nuclear Lesion-induced Synaptic Reorganization in the Basilar Pons of the Rat (대뇌피질 또는 소뇌핵 병변에 따른 쥐 교핵내 연접구조의 변화)

  • 이현숙
    • The Korean Journal of Zoology
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    • v.34 no.3
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    • pp.382-388
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    • 1991
  • 쥐의 교핵내에서 대뇌피질(또는 소뇌핵) 면면시 소뇌교핵계(또는 대뇌교핵계)의 연접구조의 변화에 대한 전자현미경적 관찰결과, 대뇌교핵계의 수입로 차단의 경우 소뇌교핵계의 신경섬유는 교핵세포의 원위 가지돌기쪽으로 발아하며, 소뇌핵 병변시 서뇌교핵계의 신경섬유는 근위가지돌기와 연접을 형성하거나 또는 여러개의 가지돌기 부속물들과 사구체형 연접복합체를 형성함이 밝혀졌다. 이상의 연구는 광학현미경적 관찰에서 언급된 대뇌피질(또는 소뇌핵) 병변에 따른 교핵내 신경종말의 밀도증가에 대한 보고를 뒷받침해주며, 소뇌핵 병변시 운동결핍증의 시간경과에 따른 회복에 대한 신경해부학적 근거를 제시한다.

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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.

Promotion of Synaptic Maturation by Deep Seawater in Cultured Rat Hippocampal Neurons (해양심층수의 해마신경세포 연접형성 촉진 효과)

  • Kim, Seong-Ho;Lee, Hyun-Sook;Shon, Yun-Hee;Nam, Kyung-Soo;Moon, Il-Soo
    • Journal of Life Science
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    • v.18 no.11
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    • pp.1479-1484
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    • 2008
  • Deep seawater (DSW) refers to water extracted from the ocean, usually at depths of 200 meters or more, which is rich in inorganic materials and has attracted attention for various applications. We investigated the effects of the DSW on the synaptic maturation of cultured rat hippocampal neurons. Immunocytochemical examination of DIV21 showed that PSD-95, $\alpha$CaMKII, and synGAP$\alpha1$clusters were strengthened and coupling rates of SV2 and NR2B were significantly increased in neurons grown in the presence of H-800 and H-1000 DSW. Our results indicate that DSW promotes the formation of excitatory postsynaptic signal transduction complexes NRC/MASC and functional synapses.