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

hPAX-P2S （human placental antigen X-P2S, hPAX-P2S）最初通过免疫组化检测到的stimoid body （STB）是一种独特的非膜结合的神经细胞质包涵体，它也含有huntingtin-associated protein 1 (HAP1A/B)，转染HAP1A- cdna可在培养细胞中诱导STB样包涵体。大量stb优先分布在很少发生神经变性的边缘和下丘脑区域，而不是纹状体、丘脑和新皮层等神经变性的靶点。目前的研究首先提供了明确的证据，证明HAP1通过其配体结合域与脊髓-球-肌萎缩（SBMA）来源的雄激素受体（AR）以多q -长度依赖的方式相互作用，形成突出的包裹体，分离多q -AR，并且双氢睾酮的加入比ARQ65更显著地降低了HAP1与ARQ25的关联强度。此外，sbma - mutt - arq65诱导的apop…通过与HAP1共转染抑制更多的tosis，支持HAP1/STB对神经退行性变起保护作用的“HAP1/STB保护假说”。此外，在Western blotting中，抗hpax - p2s抗血清首次被证明可以识别HAP1A/B c端附近的HAPI^<474-577> （XP2S结构域），并且hpax - p2s -脑STB和HAP1A诱导的包涵体的免疫反应被HAP1^<474-577>预吸附消除。这些发现清楚地表明hPAX-P2S与stb构成的HAP1相同，并且HAP1诱导的/免疫反应性包涵体对应于hPAX-P2S免疫反应性stb。此外，抗hpax - p2s抗血清对hap1b诱导的/免疫反应性弥漫性结构的免疫染色较弱，提示XP2S结构域在弥漫性结构中被某些分子覆盖，并在STB中公开，这对阐明STB形成机制具有重要线索。在免疫电镜下，HAP1不仅定位于STB，而且与STB附近的内质网状管状结构有关，但STB目前尚未被任何针对细胞器标记物（包括KDEL， GMP130, LAMP1， EEA1或Bcl2）的抗体检测到。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