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Quantitative ultrastructural localization of voltage-gated calcium channel subunits in the mouse brain
https://ir.soken.ac.jp/records/3599
https://ir.soken.ac.jp/records/35994bf8ae13-4b67-4dc2-9a47-b8adf6b2bcee
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要旨・審査要旨 (313.5 kB)
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本文 (2.4 MB)
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| Item type | 学位論文 / Thesis or Dissertation(1) | |||||||
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| 公開日 | 2013-06-18 | |||||||
| タイトル | ||||||||
| タイトル | Quantitative ultrastructural localization of voltage-gated calcium channel subunits in the mouse brain | |||||||
| タイトル | ||||||||
| タイトル | Quantitative ultrastructural localization of voltage-gated calcium channel subunits in the mouse brain | |||||||
| 言語 | en | |||||||
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| 言語 | eng | |||||||
| 資源タイプ | ||||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_46ec | |||||||
| 資源タイプ | thesis | |||||||
| 著者名 |
PARAJULI, Laxmi Kumar
× PARAJULI, Laxmi Kumar
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| フリガナ |
バラジュリ, ラクシミ クマール
× バラジュリ, ラクシミ クマール
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| 著者 |
PARAJULI, Laxmi Kumar
× PARAJULI, Laxmi Kumar
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| 学位授与機関 | ||||||||
| 学位授与機関名 | 総合研究大学院大学 | |||||||
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| 学位名 | 博士（理学） | |||||||
| 学位記番号 | ||||||||
| 内容記述タイプ | Other | |||||||
| 内容記述 | 総研大甲第1565号 | |||||||
| 研究科 | ||||||||
| 値 | 生命科学研究科 | |||||||
| 専攻 | ||||||||
| 値 | 20 生理科学専攻 | |||||||
| 学位授与年月日 | ||||||||
| 学位授与年月日 | 2012-09-28 | |||||||
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| 値 | 2012 | |||||||
| 要旨 | ||||||||
| 内容記述タイプ | Other | |||||||
| 内容記述 | My thesis deals with the localization of T-type calcium channel are R-type calcium channel in the brain. T-type calcium channels play a pivotal role in regulating neural membrane excitability. However, the precise subcellular distributions of T-type channel subunits are not well understood. Here, I investigated the subcellular distribution of the α1G subunit of the calcium channel which is expressed highly in the mouse dorsal lateral geniculate nucleus (dLGN). Light microscopic analysis demonstrated that dLGN exhibits intense immunoperoxidase reactivity for the α1G subunit. Electron microscopic observation showed that the labeling was present in both the relay cells and interneurons and was found in the somatodendritic, but not axonal, domains of these cells. Most of the immunogold particles for the α1G subunit were either associated with the plasma membrane or the intracellular membranes. Reconstruction analysis of serial electron microscopic images revealed that the intensity of the intracellular labeling exhibited gradient such that the labeling density was the highest in the proximal dendrite and progressively decreased towards the distal dendrite. In contrast, the plasma membrane associated particles were distributed with a uniform density over the somatodendritic surface of dLGN cells. The labeling density in the relay cell plasma membrane was about three-fold higher than that of the interneurons. These results provide ultrastructural evidence for cell-type specific expression levels and for uniform expression density of the α1G subunit over the plasma membrane of dLGN cells. R-type calcium channels (RTCC) are well known for their role in synaptic plasticity, but little is known about their subcellular distribution across various neuronal compartments. Using subtype-specific antibodies, I characterized the regional and subcellular localization of Cav2.3 at both light and electron microscopic levels. Cav2.3 immunogold particles were found to be predominantly presynaptic in the interpeduncular nucleus, but postsynaptic in other brain regions. Serial section analysis of electron microscopic images from the hippocampal CA1 revealed a higher density of immunogold particles in the dendritic shaft plasma membrane compared with the pyramidal cell somata. However, the labeling densities were not significantly different among the apical, oblique or basal dendrites. Immunogold particles were also observed over the plasma membrane of dendritic spines, including both synaptic and extrasynaptic sites. Individual spine heads contained < 20 immunogold particles, with an average density of 260 immunoparticles per µm3 spine head volume, in accordance with the density of RTCCs estimated using calcium imaging (Sabatini and Svoboda, 2000). The Cav2.3 density was variable among similar-sized spine heads and did not correlate with the density in the parent dendrite, implying that spines are individual calcium compartments operating autonomously from their parent dendrites. |
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| 値 | 有 | |||||||
