Ultrastructural analysis and molecular screening of the neuronal cytoplasmic inclusion, "the stigmoid body"

神经元细胞质内含物“柱状体”的超微结构分析和分子筛选

• 批准号: 17500231
• 负责人: SHINODA Koh
• 金额: $ 1.86万
• 依托单位: Yamaguchi University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17500231/
• 关键词: stigmoid body; hPAX-P2; HAP1; spinal and bulbar muscular atrophy; Huntington's disease; androgen receptor; endoplasmic reticulum; neurocytoplasmic inclusion; sortilin; SorLA; LR11; tublin; dynactin; エストロゲン受容体; mRNA; 神経細胞; 視床下部; 脊髄球筋萎縮症

The stigmoid body (STB), which was a distinct non-membrane-bound neurocytoplasmic inclusion originally detected by immunohistochemistry for human placental antigen X-P2S (hPAX-P2S), also contains huntingtin-associated protein 1 (HAP1A/B), and the HAP1A-cDNA transfection induces STB-like inclusions in cultured cells. A striking number of the STBs are preferentially distributed in the limbic and hypothalamic regions, where neurodegeneration rarely occurs, rather than the targets of neurodegeneration including the striatum, thalamus and neocortex. The current research first provided clear evidence that HAP1 interacts with androgen receptor (AR) derived from spinal-and-bulbar-muscular-atrophy (SBMA) via its ligand-binding domain in a polyQ-length-dependent manner and forms prominent inclusions sequestering polyQ-AR, and that addition of dihydrotestosterone reduces the association strength of HAP1 with ARQ25 more dramatically than that with ARQ65. Furthermore, SBMA-mutant-ARQ65-induced apop … More tosis was suppressed by cotransfection with HAP1, supporting "the HAP1/STB protection hypothesis" that the HAP1/STB plays a protective role against neurodegeneration.In addition, the anti-hPAX-P2S antiserum was first demonstrated to recognize HAPI^<474-577> (XP2S domain) near C-terminuses of HAP1A/B in Western blotting, and hPAX-P2S-immunoreactions of the brain STB and HAP1A-induced inclusions were shown to be eliminated by pre-adsorption with HAP1^<474-577>. These findings clearly indicated that hPAX-P2S is identical with STB-constituted HAP1 and that the HAP1-induced/immunoreactive inclusions correspond to the hPAX-P2S-immunoreactive STBs. In addition, the anti-hPAX-P2S antiserum far weakly immunostained HAP1B-induced/immunoreactive diffuse structures, suggesting that the XP2S domain is covered by some molecule in the diffuse structures and disclosed in the STB and have an important clue to elucidate mechanism of the STB formation. In immuno-electron microscopy, HAP1 is not only localized to the STB but also associated with endoplasmic-reticulum-like tubular structures adjacent to the STB, but the STB is not detected so far by any antibodies to organelle-markers including KDEL, GMP130, LAMP1, EEA1 or Bcl2. The organelle origin has yet to be determined. Less

最初通过人胎盘抗原X-P2 S（hPAX-P2 S）免疫组织化学检测到的豆状体（STB）是一种独特的非膜结合的神经细胞质包涵体，也含有亨廷顿相关蛋白1（HAP 1A/B），并且HAP 1A-cDNA转染在培养的细胞中诱导STB样包涵体。显著数量的STB优先分布在边缘系统和下丘脑区域，在那里很少发生神经变性，而不是包括纹状体，丘脑和新皮质在内的神经变性的靶点。目前的研究首次提供了明确的证据表明，HAP 1通过其配体结合结构域以polyQ长度依赖的方式与来自脊髓延髓肌萎缩症（SBMA）的雄激素受体（AR）相互作用，并形成突出的包含物隔离polyQ-AR，并且添加双氢睾酮降低HAP 1与ARQ 25的缔合强度比与ARQ 65的缔合强度更显著。此外，SBMA突变体ARQ 65诱导的apo ...更多信息 HAP 1共转染抑制了HAP 1的凋亡，支持了HAP 1/STB保护假说，即HAP 1/STB对神经退行性变起保护作用。<474-577>蛋白质印迹法中HAP 1A/B C末端附近的XP 2S结构域，脑STB和HAP 1A诱导的包涵体的hPAX-P2 S免疫反应显示通过用HAP 1A预吸附而消除<474-577>。这些发现清楚地表明hPAX-P2 S与STB构成的HAP 1相同，并且HAP 1诱导的/免疫反应性包涵体对应于hPAX-P2 S免疫反应性STB。另外，抗hPAX-P2 S抗血清对HAP 1B诱导的免疫反应性弥散结构的免疫染色较弱，提示XP 2S结构域被弥散结构中的某些分子所覆盖，并在STB中被揭示，对阐明STB的形成机制具有重要的线索。在免疫电子显微镜中，HAP 1不仅定位于STB，而且还与STB附近的内质网样管状结构相关，但迄今为止，STB未被针对细胞器标记物（包括KDEL、GMP 130、LAMP 1、EEA 1或Bcl 2）的任何抗体检测到。细胞器的起源尚未确定。少

项目成果

Region specific expression and sex steroidal regulation on aromatase and its mRNA in the male rat brain

雄性大鼠脑中芳香酶及其mRNA的区域特异性表达和性别类固醇调节

• 发表时间: 2007
• 作者: Kokubu;K.;et. al.
• 通讯作者: et. al.

Sex-Steroidal Regulation on Region-Specific Expression of Aromatase in the Male Rat Brain

雄性大鼠脑中芳香酶区域特异性表达的性别类固醇调节

• 发表时间: 2006
• 作者: Kokubu;K.;et. al.
• 通讯作者: et. al.

Ultrastructural analysis of subcellular expression of huntingtin associated protein 1 and formation of the stigmoid body

亨廷顿蛋白相关蛋白1亚细胞表达及柱状体形成的超微结构分析

• 发表时间: 2006
• 作者: Yanai;A.;et. al.
• 通讯作者: et. al.

Sex-steroidal regulation of aromatase mRNA expression in adult male rat brain: a quantitative non-radioactive in situ hybridization study

• DOI: 10.1007/s00441-008-0606-8
• 发表时间: 2008-04
• 期刊: Cell and Tissue Research
• 影响因子: 3.6
• 作者: Changjiu Zhao;Ryutaro Fujinaga;Akie Yanai;K. Kokubu;Y. Takeshita;Yoshifumi Watanabe;K. Shinoda

Fibroblast growth factor 2 facilitates the differentiation of transplanted bone marrow cells into hepatocytes

• DOI: 10.1007/s00441-005-0077-0
• 发表时间: 2006-02-01
• 期刊: CELL AND TISSUE RESEARCH
• 影响因子: 3.6
• 作者: Ishikawa, T;Terai, S;Okita, K
• 通讯作者: Okita, K