2021/09/20 更新

写真a

ミヤザキ ハルコ
宮﨑 晴子
Miyazaki Haruko
所属
医歯薬学域 助教
職名
助教
外部リンク

学位

  • 博士(医学) ( 筑波大学 )

研究キーワード

  • 無髄神経

  • 有髄神経

  • ミエリン

  • ハンチントン病

  • 大脳基底核

  • 神経変性疾患

  • ナトリウムチャネル

  • 神経科学

研究分野

  • ライフサイエンス / 病態神経科学  / ナトリウムチャネル

  • ライフサイエンス / 病態神経科学  / 髄鞘

  • ライフサイエンス / 神経内科学  / 大脳基底核

  • ライフサイエンス / 病態神経科学  / ハンチントン病

  • ライフサイエンス / 神経内科学  / ハンチントン病

学歴

  • 筑波大学    

    2005年4月 - 2008年3月

      詳細を見る

  • 筑波大学    

    2003年4月 - 2005年3月

      詳細を見る

  • 麻布大学   獣医学部   動物応用科学科

    1995年4月 - 1999年3月

      詳細を見る

経歴

  • 岡山大学   大学院医歯薬学総合研究科 分子医化学分野   助教

    2021年5月 - 現在

      詳細を見る

    国名:日本国

    researchmap

  • 同志社大学   脳科学研究科 認知記憶加齢部門   助教

    2014年4月 - 2021年4月

      詳細を見る

  • 理化学研究所   脳科学総合研究センター 構造神経病理研究チーム   研究員

    2008年4月 - 2013年3月

      詳細を見る

  • 理化学研究所   脳科学総合研究センター 構造神経病理研究チーム   ジュニアリサーチアソシエイト

    2005年4月 - 2008年3月

      詳細を見る

  • 理化学研究所   脳科学総合研究センター 構造神経病理研究チーム   テクニカルスタッフ

    2001年1月 - 2005年3月

      詳細を見る

  • 理化学研究所   脳科学総合研究センター シナプス分子機構研究チーム   テクニカルスタッフ

    1999年4月 - 2000年12月

      詳細を見る

▼全件表示

 

論文

  • FACS-array-based cell purification yields a specific transcriptome of striatal medium spiny neurons in a murine Huntington disease model. 査読 国際誌

    Haruko Miyazaki, Tomoyuki Yamanaka, Fumitaka Oyama, Yoshihiro Kino, Masaru Kurosawa, Mizuki Yamada-Kurosawa, Risa Yamano, Tomomi Shimogori, Nobutaka Hattori, Nobuyuki Nukina

    The Journal of biological chemistry   295 ( 29 )   9768 - 9785   2020年7月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Huntington disease (HD) is a neurodegenerative disorder caused by expanded CAG repeats in the Huntingtin gene. Results from previous studies have suggested that transcriptional dysregulation is one of the key mechanisms underlying striatal medium spiny neuron (MSN) degeneration in HD. However, some of the critical genes involved in HD etiology or pathology could be masked in a common expression profiling assay because of contamination with non-MSN cells. To gain insight into the MSN-specific gene expression changes in presymptomatic R6/2 mice, a common HD mouse model, here we used a transgenic fluorescent protein marker of MSNs for purification via FACS before profiling gene expression with gene microarrays and compared the results of this "FACS-array" with those obtained with homogenized striatal samples (STR-array). We identified hundreds of differentially expressed genes (DEGs) and enhanced detection of MSN-specific DEGs by comparing the results of the FACS-array with those of the STR-array. The gene sets obtained included genes ubiquitously expressed in both MSNs and non-MSN cells of the brain and associated with transcriptional regulation and DNA damage responses. We proposed that the comparative gene expression approach using the FACS-array may be useful for uncovering the gene cascades affected in MSNs during HD pathogenesis.

    DOI: 10.1074/jbc.RA120.012983

    PubMed

    researchmap

  • Singular localization of sodium channel β4 subunit in unmyelinated fibres and its role in the striatum. 査読

    Miyazaki H, Oyama F, Inoue R, Aosaki T, Abe T, Kiyonari H, Kino Y, Kurosawa M, Shimizu J, Ogiwara I, Yamakawa K, Koshimizu Y, Fujiyama F, Kaneko T, Shimizu H, Nagatomo K, Yamada K, Shimogori T, Hattori N, Miura M, Nukina N

    Nature communications   5   5525   2014年11月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1038/ncomms6525

    Web of Science

    PubMed

    researchmap

  • Establishment of Feeder-Independent Cloned Caprine Trophoblast Cell Line Which Expresses Placental Lactogen and Interferon Tau 査読

    H. Miyazaki, M. Imai, T. Hirayama, S. Saburi, M. Tanaka, M. Maruyama, C. Matsuo, H. Meguro, K. Nishibashi, F. Inoue, J. Djiane, A. Gertler, S. Tachi, K. Imakawa, C. Tachi

    Placenta   23 ( 8-9 )   613 - 630   2002年9月

     詳細を見る

    掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier BV  

    DOI: 10.1053/plac.2002.0846

    researchmap

  • Proteomic analysis of subcellular compartments containing disseminated alpha-synuclein seeds. 国際誌

    Junya Kasahara, Yukio Imamura, Akiko Hiyama, Tomoyuki Yamanaka, Haruko Miyazaki, Nobuyuki Nukina

    Neuroscience research   2020年12月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The pathological form of a-synuclein (a-syn) is transmitted through neural circuits in the brains of Parkinson disease (PD) patients and amplifies misfolded a-syn, further forming intracellular deposits. However, the details of a-syn pre-formed fibrils (PFFs) transmission in vivo have not been fully elucidated. By inoculating Quantum dots (QD)-labeled a-syn PFFs (QD-a-syn PFFs) into the unilateral striatum, we detected QD-a-syn PFFs in brain homogenates obtained from the ipsilateral and contralateral sides of the inoculated site and further obtained QD-a-syn PFFs enriched-particles with fluorescence-activated organelle sorting. Proteomic analysis suggested that QD-a-syn PFFs-enriched particles in the contralateral side were associated with component proteins of synapse. In contrast, QD-a-syn PFFs-enriched particles in the ipsilateral side were associated with proteins belonging to ER components. Immunostaining of brain sections confirmed that QD-a-syn PFFs in the contralateral side were co-localized with synaptic vesicle marker proteins in the cortex and striatum. Additionally, QD-a-syn PFFs in the ipsilateral side were more co-localized with ER marker proteins compared to the contralateral side. These results correspond to proteomic analysis. This study provides potential candidates for the subcellular localization of a-syn PFFs in vivo during the dissemination phase of seeds. These subcellular compartments could be involved in the transmission of seeds.

    DOI: 10.1016/j.neures.2020.11.009

    PubMed

    researchmap

  • Gene expression profiling in neuronal cells identifies a different type of transcriptome modulated by NF-Y. 国際誌

    Tomoyuki Yamanaka, Haruko Miyazaki, Asako Tosaki, Sankar N Maity, Tomomi Shimogori, Nobutaka Hattori, Nobuyuki Nukina

    Scientific reports   10 ( 1 )   21714 - 21714   2020年12月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A heterotrimeric transcription factor NF-Y is crucial for cell-cycle progression in various types of cells. In contrast, studies using NF-YA knockout mice have unveiled its essential role in endoplasmic reticulum (ER) homeostasis in neuronal cells. However, whether NF-Y modulates a different transcriptome to mediate distinct cellular functions remains obscure. Here, we knocked down NF-Y in two types of neuronal cells, neuro2a neuroblastoma cells and mouse brain striatal cells, and performed gene expression profiling. We found that down-regulated genes preferentially contained NF-Y-binding motifs in their proximal promoters, and notably enriched genes related to ER functions rather than those for cell cycle. This contrasts with the profiling data of HeLa and embryonic stem cells in which distinct down-regulation of cell cycle-related genes was observed. Clustering analysis further identified several functional clusters where populations of the down-regulated genes were highly distinct. Further analyses using chromatin immunoprecipitation and RNA-seq data revealed that the transcriptomic difference was not correlated with DNA binding of NF-Y but with splicing of NF-YA. These data suggest that neuronal cells have a different type of transcriptome in which ER-related genes are dominantly modulated by NF-Y, and imply that NF-YA splicing alteration could be involved in this cell type-specific gene modulation.

    DOI: 10.1038/s41598-020-78682-8

    PubMed

    researchmap

  • Preserved proteinase K-resistant core after amplification of alpha-synuclein aggregates: Implication to disease-related structural study. 査読 国際誌

    Saki Yoshinaga, Tomoyuki Yamanaka, Haruko Miyazaki, Ayami Okuzumi, Akiko Hiyama, Shigeo Murayama, Nobuyuki Nukina

    Biochemical and biophysical research communications   522 ( 3 )   655 - 661   2020年2月

     詳細を見る

    記述言語:英語  

    Many pathological proteins related to neurodegenerative diseases are misfolded, aggregating to form amyloid fibrils during pathogenesis. One of the pathological proteins, alpha-synuclein (α-syn), accumulates in the brains of Parkinson disease (PD), dementia with Lewy bodies (DLB) and multiple system atrophy (MSA), which are designated as synucleinopathies. Recently, structural properties of abnormal accumulated proteins are suggested to determine the disease phenotype. However, the biochemical and structural characteristics of those accumulated proteins are still poorly understood. We previously reported the sequence and seed-structure-dependent polymorphic fibrils of α-syn and the polymorphism was identified by proteinase K-resistant cores determined by mass spectrometry (MS) analysis. In this study, we applied this method to analyze α-syn aggregates of MSA and DLB. To perform MS analysis on proteinase K-resistant cores, we first performed amplification of α-syn aggregates by seeding reaction and protein misfolding cyclic amplification (PMCA) to obtain a sufficient amount of aggregates. Using SDS insoluble fraction of the disease brain, we successfully amplified enough α-syn aggregates for MS analysis. We differentiated between mouse and human α-syn aggregates by MS analysis on proteinase K-resistant cores of the aggregates before and after amplification. The results suggest that structural properties of amplified α-syn fibrils are preserved after PMCA and these methods can be applicable in the study of pathological proteins of the neurodegenerative disorders.

    DOI: 10.1016/j.bbrc.2019.11.142

    PubMed

    researchmap

  • Non-coding RNA Neat1 and Abhd11os expressions are dysregulated in medium spiny neurons of Huntington disease model mice. 査読 国際誌

    Hongsun Park, Haruko Miyazaki, Tomoyuki Yamanaka, Nobuyuki Nukina

    Neuroscience research   147   58 - 63   2019年10月

     詳細を見る

    記述言語:英語  

    Huntington Disease (HD) is a neurodegenerative disorder caused by expanded CAG repeats in the exon1 of huntingtin gene (HTT). The mutant HTT affects the transcriptional profile of neurons by disrupting the activities of transcriptional machinery and alters expression of many genes. In this study, we identified dysregulated non-coding RNAs (ncRNAs) in medium spiny neurons of 4-week-old HD model mouse. Also, we observed the intracellular localizations of Abhd11os and Neat1 ncRNAs by ViewRNA in situ hybridization, which could provide more precise detection, suggesting that it is a useful method to investigate the expression changes of genes with low expression levels.

    DOI: 10.1016/j.neures.2018.10.013

    PubMed

    researchmap

  • Rapid dissemination of alpha-synuclein seeds through neural circuits in an in-vivo prion-like seeding experiment. 査読 国際誌

    Ayami Okuzumi, Masaru Kurosawa, Taku Hatano, Masashi Takanashi, Shuuko Nojiri, Takeshi Fukuhara, Tomoyuki Yamanaka, Haruko Miyazaki, Saki Yoshinaga, Yoshiaki Furukawa, Tomomi Shimogori, Nobutaka Hattori, Nobuyuki Nukina

    Acta neuropathologica communications   6 ( 1 )   96 - 96   2018年9月

     詳細を見る

    記述言語:英語  

    Accumulating evidence suggests that the lesions of Parkinson's disease (PD) expand due to transneuronal spreading of fibrils composed of misfolded alpha-synuclein (a-syn), over the course of 5-10 years. However, the precise mechanisms and the processes underlying the spread of these fibril seeds have not been clarified in vivo. Here, we investigated the speed of a-syn transmission, which has not been a focus of previous a-syn transmission experiments, and whether a-syn pathologies spread in a neural circuit-dependent manner in the mouse brain. We injected a-syn preformed fibrils (PFFs), which are seeds for the propagation of a-syn deposits, either before or after callosotomy, to disconnect bilateral hemispheric connections. In mice that underwent callosotomy before the injection, the propagation of a-syn pathology to the contralateral hemisphere was clearly reduced. In contrast, mice that underwent callosotomy 24 h after a-syn PFFs injection showed a-syn pathology similar to that seen in mice without callosotomy. These results suggest that a-syn seeds are rapidly disseminated through neuronal circuits immediately after seed injection, in a prion-like seeding experiment in vivo, although it is believed that clinical a-syn pathologies take years to spread throughout the brain. In addition, we found that botulinum toxin B blocked the transsynaptic transmission of a-syn seeds by specifically inactivating the synaptic vesicle fusion machinery. This study offers a novel concept regarding a-syn propagation, based on the Braak hypothesis, and also cautions that experimental transmission systems may be examining a unique type of transmission, which differs from the clinical disease state.

    DOI: 10.1186/s40478-018-0587-0

    PubMed

    researchmap

  • Parallel homodimer structures of the extracellular domains of the voltage-gated sodium channel β4 subunit explain its role in cell-cell adhesion. 査読

    Shimizu H, Tosaki A, Ohsawa N, Ishizuka-Katsura Y, Shoji S, Miyazaki H, Oyama F, Terada T, Shirouzu M, Sekine SI, Nukina N, Yokoyama S

    The Journal of biological chemistry   292 ( 32 )   13428 - 13440   2017年8月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1074/jbc.M117.786509

    Web of Science

    PubMed

    researchmap

  • Acidic mammalian chitinase is a proteases-resistant glycosidase in mouse digestive system 査読

    Misa Ohno, Masahiro Kimura, Haruko Miyazaki, Kazuaki Okawa, Riho Onuki, Chiyuki Nemoto, Eri Tabata, Satoshi Wakita, Akinori Kashimura, Masayoshi Sakaguchi, Yasusato Sugahara, Nobuyuki Nukina, Peter O. Bauer, Fumitaka Oyama

    SCIENTIFIC REPORTS   6   37756   2016年11月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Chitinases are enzymes that hydrolyze chitin, a polymer of beta-1, 4-linked N-acetyl-D-glucosamine (GlcNAc). Chitin has long been considered as a source of dietary fiber that is not digested in the mammalian digestive system. Here, we provide evidence that acidic mammalian chitinase (AMCase) can function as a major digestive enzyme that constitutively degrades chitin substrates and produces (GlcNAc) 2 fragments in the mouse gastrointestinal environment. AMCase was resistant to endogenous pepsin C digestion and remained active in the mouse stomach extract at pH 2.0. The AMCase mRNA levels were much higher than those of four major gastric proteins and two housekeeping genes and comparable to the level of pepsinogen C in the mouse stomach tissues. Furthermore, AMCase was expressed in the gastric pepsinogen-synthesizing chief cells. The enzyme was also stable and active in the presence of trypsin and chymotrypsin at pH 7.6, where pepsin C was completely degraded. Mouse AMCase degraded polymeric colloidal and crystalline chitin substrates in the gastrointestinal environments in presence of the proteolytic enzymes. Thus, AMCase can function as a protease-resistant major glycosidase under the conditions of stomach and intestine and degrade chitin substrates to produce (GlcNAc) 2, a source of carbon, nitrogen and energy.

    DOI: 10.1038/srep37756

    Web of Science

    PubMed

    researchmap

  • Differential roles of NF-Y transcription factor in ER chaperone expression and neuronal maintenance in the CNS 査読

    Tomoyuki Yamanaka, Asako Tosaki, Haruko Miyazaki, Masaru Kurosawa, Masato Koike, Yasuo Uchiyama, Sankar N. Maity, Hidemi Misawa, Ryosuke Takahashi, Tomomi Shimogori, Nobutaka Hattori, Nobuyuki Nukina

    SCIENTIFIC REPORTS   6   34575   2016年9月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    The mammalian central nervous system (CNS) contains various types of neurons with different neuronal functions. In contrast to established roles of cell type-specific transcription factors on neuronal specification and maintenance, whether ubiquitous transcription factors have conserved or differential neuronal function remains uncertain. Here, we revealed that inactivation of a ubiquitous factor NF-Y in different sets of neurons resulted in cell type-specific neuropathologies and gene downregulation in mouse CNS. In striatal and cerebellar neurons, NF-Y inactivation led to ubiquitin/p62 pathologies with downregulation of an endoplasmic reticulum (ER) chaperone Grp94, as we previously observed by NF-Y deletion in cortical neurons. In contrast, NF-Y inactivation in motor neurons induced neuronal loss without obvious protein deposition. Detailed analysis clarified downregulation of another ER chaperone Grp78 in addition to Grp94 in motor neurons, and knockdown of both ER chaperones in motor neurons recapitulated the pathology observed after NF-Y inactivation. Finally, additional downregulation of Grp78 in striatal neurons suppressed ubiquitin accumulation induced by NF-Y inactivation, implying that selective ER chaperone downregulation mediates different neuropathologies. Our data suggest distinct roles of NF-Y in protein homeostasis and neuronal maintenance in the CNS by differential regulation of ER chaperone expression.

    DOI: 10.1038/srep34575

    Web of Science

    PubMed

    researchmap

  • Structure-based site-directed photo-crosslinking analyses of multimeric cell-adhesive interactions of voltage-gated sodium channel β subunits. 査読

    Shimizu H, Miyazaki H, Ohsawa N, Shoji S, Ishizuka-Katsura Y, Tosaki A, Oyama F, Terada T, Sakamoto K, Shirouzu M, Sekine S, Nukina N, Yokoyama S

    Scientific reports   6   26618   2016年5月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1038/srep26618

    Web of Science

    PubMed

    researchmap

  • Proteomic analysis of native cerebellar iFGF14 complexes 査読

    Marie K. Bosch, Jeanne M. Nerbonne, R. Reid Townsend, Haruko Miyazaki, Nobuyuki Nukina, David M. Ornitz, Celine Marionneau

    CHANNELS   10 ( 4 )   297 - 312   2016年

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS INC  

    Intracellular Fibroblast Growth Factor 14 (iFGF14) and the other intracellular FGFs (iFGF11-13) regulate the properties and densities of voltage-gated neuronal and cardiac Na+ (Nav) channels. Recent studies have demonstrated that the iFGFs can also regulate native voltage-gated Ca2+ (Cav) channels. In the present study, a mass spectrometry (MS)-based proteomic approach was used to identify the components of native cerebellar iFGF14 complexes. Using an anti-iFGF14 antibody, native iFGF14 complexes were immunoprecipitated from wild type adult mouse cerebellum. Parallel control experiments were performed on cerebellar proteins isolated from mice (Fgf14(-/-)) harboring a targeted disruption of the Fgf14 locus. MS analyses of immunoprecipitated proteins demonstrated that the vast majority of proteins identified in native cerebellar iFGF14 complexes are Nav channel pore-forming () subunits or proteins previously reported to interact with Nav subunits. In contrast, no Cav channel or accessory subunits were revealed in cerebellar iFGF14 immunoprecipitates. Additional experiments were completed using an anti-PanNav antibody to immunoprecipitate Nav channel complexes from wild type and Fgf14(-/-) mouse cerebellum. Western blot and MS analyses revealed that the loss of iFGF14 does not measurably affect the protein composition or the relative abundance of Nav channel interacting proteins in native adult mouse cerebellar Nav channel complexes.

    DOI: 10.1080/19336950.2016.1153203

    Web of Science

    researchmap

  • Sodium channel β1 subunit localizes to axon initial segments of excitatory and inhibitory neurons and shows regional heterogeneity in mouse brain. 査読

    Wimmer VC, Harty RC, Richards KL, Phillips AM, Miyazaki H, Nukina N, Petrou S

    The Journal of comparative neurology   523 ( 5 )   814 - 830   2015年4月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1002/cne.23715

    Web of Science

    PubMed

    researchmap

  • FUS/TLS deficiency causes behavioral and pathological abnormalities distinct from amyotrophic lateral sclerosis 査読

    Yoshihiro Kino, Chika Washizu, Masaru Kurosawa, Mizuki Yamada, Haruko Miyazaki, Takumi Akagi, Tsutomu Hashikawa, Hiroshi Doi, Toru Takumi, Geoffrey G. Hicks, Nobutaka Hattori, Tomomi Shimogori, Nobuyuki Nukina

    ACTA NEUROPATHOLOGICA COMMUNICATIONS   3   24   2015年4月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BIOMED CENTRAL LTD  

    Introduction: FUS/TLS is an RNA-binding protein whose genetic mutations or pathological inclusions are associated with neurological diseases including amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration, and essential tremor (ET). It is unclear whether their pathogenesis is mediated by gain or loss of function of FUS/TLS.
    Results: Here, we established outbred FUS/TLS knockout mice to clarify the effects of FUS/TLS dysfunction in vivo. We obtained homozygous knockout mice that grew into adulthood. Importantly, they did not manifest ALS-or ET-like phenotypes until nearly two years. Instead, they showed distinct histological and behavioral alterations including vacuolation in hippocampus, hyperactivity, and reduction in anxiety-like behavior. Knockout mice showed transcriptome alterations including upregulation of Taf15 and Hnrnpa1, while they have normal morphology of RNA-related granules such as Gems.
    Conclusions: Collectively, FUS/TLS depletion causes phenotypes possibly related to neuropsychiatric and neurodegenerative conditions, but distinct from ALS and ET, together with specific alterations in RNA metabolisms.

    DOI: 10.1186/s40478-015-0202-6

    Web of Science

    PubMed

    researchmap

  • Modulation of voltage-gated K+ channels by the sodium channel β1 subunit. 査読

    Nguyen HM, Miyazaki H, Hoshi N, Smith BJ, Nukina N, Goldin AL, Chandy KG

    Proceedings of the National Academy of Sciences of the United States of America   109 ( 45 )   18577 - 18582   2012年11月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1073/pnas.1209142109

    Web of Science

    PubMed

    researchmap

  • Mutant huntingtin fragment selectively suppresses Brn-2 POU domain transcription factor to mediate hypothalamic cell dysfunction 査読

    Tomoyuki Yamanaka, Asako Tosaki, Haruko Miyazaki, Masaru Kurosawa, Yoshiaki Furukawa, Mizuki Yamada, Nobuyuki Nukina

    HUMAN MOLECULAR GENETICS   19 ( 11 )   2099 - 2112   2010年6月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    In polyglutamine diseases including Huntington's disease (HD), mutant proteins containing expanded polyglutamine stretches form nuclear aggregates in neurons. Although analysis of their disease models suggested a significance of transcriptional dysregulation in these diseases, how it mediates the specific neuronal cell dysfunction remains obscure. Here we performed a comprehensive analysis of altered DNA binding of multiple transcription factors using R6/2 HD model mice brains that express an N-terminal fragment of mutant huntingtin (mutant Nhtt). We found a reduction of DNA binding of Brn-2, a POU domain transcription factor involved in differentiation and function of hypothalamic neurosecretory neurons. We provide evidence supporting that Brn-2 loses its function through two pathways, its sequestration by mutant Nhtt and its reduced transcription, leading to reduced expression of hypothalamic neuropeptides. In contrast to Brn-2, its functionally related protein, Brn-1, was not sequestered by mutant Nhtt but was upregulated in R6/2 brain, except in hypothalamus. Our data indicate that functional suppression of Brn-2 together with a region-specific lack of compensation by Brn-1 mediates hypothalamic cell dysfunction by mutant Nhtt.

    DOI: 10.1093/hmg/ddq087

    Web of Science

    PubMed

    researchmap

  • Harnessing chaperone-mediated autophagy for the selective degradation of mutant huntingtin protein 査読

    Peter O. Bauer, Anand Goswami, Hon Kit Wong, Misako Okuno, Masaru Kurosawa, Mizuki Yamada, Haruko Miyazaki, Gen Matsumoto, Yoshihiro Kino, Yoshitaka Nagai, Nobuyuki Nukina

    NATURE BIOTECHNOLOGY   28 ( 3 )   256 - U111   2010年3月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Huntington's Disease (HD) is a dominantly inherited pathology caused by the accumulation of mutant huntingtin protein (HTT) containing an expanded polyglutamine (polyQ) tract. As the polyglutamine binding peptide 1 (QBP1) is known to bind an expanded polyQ tract but not the polyQ motif found in normal HTT, we selectively targeted mutant HTT for degradation by expressing a fusion molecule comprising two copies of QBP1 and copies of two different heat shock cognate protein 70 (HSC70)-binding motifs in cellular and mouse models of HD. Chaperone-mediated autophagy contributed to the specific degradation of mutant HTT in cultured cells expressing the construct. Intrastriatal delivery of a virus expressing the fusion molecule ameliorated the disease phenotype in the R6/2 mouse model of HD. Similar adaptor molecules comprising HSC70-binding motifs fused to an appropriate structure-specific binding agent(s) may have therapeutic potential for treating diseases caused by misfolded proteins other than those with expanded polyQ tracts.

    DOI: 10.1038/nbt.1608

    Web of Science

    PubMed

    researchmap

  • Functional reciprocity between Na+ channel Na(v)1.6 and beta 1 subunits in the coordinated regulation of excitability and neurite outgrowth 査読

    William J. Brackenbury, Jeffrey D. Calhoun, Chunling Chen, Haruko Miyazaki, Nobuyuki Nukina, Fumitaka Oyama, Barbara Ranscht, Lori L. Isom

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   107 ( 5 )   2283 - 2288   2010年2月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATL ACAD SCIENCES  

    Voltage-gated Na+ channel (VGSC) beta 1 subunits regulate cell-cell adhesion and channel activity in vitro. We previously showed that beta 1 promotes neurite outgrowth in cerebellar granule neurons (CGNs) via homophilic cell adhesion, fyn kinase, and contactin. Here we demonstrate that beta 1-mediated neurite outgrowth requires Na+ current (I-Na) mediated by Na(v)1.6. In addition, beta 1 is required for high-frequency action potential firing. Transient INa is unchanged in Scn1b (beta 1) null CGNs; however, the resurgent I-Na, thought to underlie high-frequency firing in Na(v)1.6-expressing cerebellar neurons, is reduced. The proportion of axon initial segments (AIS) expressing Nav1.6 is reduced in Scn1b null cerebellar neurons. In place of Na(v)1.6 at the AIS, we observed an increase in Na(v)1.1, whereas Na(v)1.2 was unchanged. This indicates that beta 1 is required for normal localization of Nav1.6 at the AIS during the postnatal developmental switch to Na(v)1.6-mediated high-frequency firing. In agreement with this, beta 1 is normally expressed with a subunits at the AIS of P14 CGNs. We propose reciprocity of function between beta 1 and Na(v)1.6 such that beta 1-mediated neurite outgrowth requires Na(v)1.6-mediated I-Na, and Na(v)1.6 localization and consequent high-frequency firing require beta 1. We conclude that VGSC subunits function in macromolecular signaling complexes regulating both neuronal excitability and migration during cerebellar development.

    DOI: 10.1073/pnas.0909434107

    Web of Science

    PubMed

    researchmap

  • A Functional Null Mutation of SCN1B in a Patient with Dravet Syndrome 査読

    Gustavo A. Patino, Lieve R. F. Claes, Luis F. Lopez-Santiago, Emily A. Slat, Raja S. R. Dondeti, Chunling Chen, Heather A. O'Malley, Charles B. B. Gray, Haruko Miyazaki, Nobuyuki Nukina, Fumitaka Oyama, Peter De Jonghe, Lori L. Isom

    JOURNAL OF NEUROSCIENCE   29 ( 34 )   10764 - 10778   2009年8月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC NEUROSCIENCE  

    Dravet syndrome (also called severe myoclonic epilepsy of infancy) is one of the most severe forms of childhood epilepsy. Most patients have heterozygous mutations in SCN1A, encoding voltage-gated sodium channel Na(v)1.1 alpha subunits. Sodium channels are modulated by beta 1 subunits, encoded by SCN1B, a gene also linked to epilepsy. Here we report the first patient with Dravet syndrome associated with a recessive mutation in SCN1B (p.R125C). Biochemical characterization of p.R125C in a heterologous system demonstrated little to no cell surface expression despite normal total cellular expression. This occurred regardless of coexpression of Na(v)1.1 alpha subunits. Because the patient was homozygous for the mutation, these data suggest a functional SCN1B null phenotype. To understand the consequences of the lack of beta 1 cell surface expression in vivo, hippocampal slice recordings were performed in Scn1b(-/-) versus Scn1b(+/+) mice. Scn1b(-/-) CA3 neurons fired evoked action potentials with a significantly higher peak voltage and significantly greater amplitude compared with wild type. However, in contrast to the Scn1a(+/-) model of Dravet syndrome, we found no measurable differences in sodium current density in acutely dissociated CA3 hippocampal neurons. Whereas Scn1b(-/-) mice seize spontaneously, the seizure susceptibility of Scn1b(+/-) mice was similar to wild type, suggesting that, like the parents of this patient, one functional SCN1B allele is sufficient for normal control of electrical excitability. We conclude that SCN1B p.R125C is an autosomal recessive cause of Dravet syndrome through functional gene inactivation.

    DOI: 10.1523/JNEUROSCI.2475-09.2009

    Web of Science

    PubMed

    researchmap

  • Genetically Determined Differences in Sodium Current Characteristics Modulate Conduction Disease Severity in Mice With Cardiac Sodium Channelopathy 査読

    Carol Ann Remme, Brendon P. Scicluna, Arie O. Verkerk, Ahmad S. Amin, Sandra van Brunschot, Leander Beekman, Vera H. M. Deneer, Catherine Chevalier, Fumitaka Oyama, Haruko Miyazaki, Nobuyuki Nukina, Ronald Wilders, Denis Escande, Remi Houlgatte, Arthur A. M. Wilde, Hanno L. Tan, Marieke W. Veldkamp, Jacques M. T. de Bakker, Connie R. Bezzina

    CIRCULATION RESEARCH   104 ( 11 )   1283 - U112   2009年6月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:LIPPINCOTT WILLIAMS & WILKINS  

    Conduction slowing of the electric impulse that drives the heartbeat may evoke lethal cardiac arrhythmias. Mutations in SCN5A, which encodes the pore-forming cardiac sodium channel alpha subunit, are associated with familial arrhythmia syndromes based on conduction slowing. However, disease severity among mutation carriers is highly variable. We hypothesized that genetic modifiers underlie the variability in conduction slowing and disease severity. With the aim of identifying such modifiers, we studied the Scn5a(1798insD/+) mutation in 2 distinct mouse strains, FVB/N and 129P2. In 129P2 mice, the mutation resulted in more severe conduction slowing particularly in the right ventricle (RV) compared to FVB/N. Pan-genomic mRNA expression profiling in the 2 mouse strains uncovered a drastic reduction in mRNA encoding the sodium channel auxiliary subunit beta 4 (Scn4b) in 129P2 mice compared to FVB/N. This corresponded to low to undetectable beta 4 protein levels in 129P2 ventricular tissue, whereas abundant beta 4 protein was detected in FVB/N. Sodium current measurements in isolated myocytes from the 2 mouse strains indicated that sodium channel activation in myocytes from 129P2 mice occurred at more positive potentials compared to FVB/N. Using computer simulations, this difference in activation kinetics was predicted to explain the observed differences in conduction disease severity between the 2 strains. In conclusion, genetically determined differences in sodium current characteristics on the myocyte level modulate disease severity in cardiac sodium channelopathies. In particular, the sodium channel subunit beta 4 (SCN4B) may constitute a potential genetic modifier of conduction and cardiac sodium channel disease. (Circ Res. 2009; 104: 1283-1292.)

    DOI: 10.1161/CIRCRESAHA.109.194423

    Web of Science

    PubMed

    researchmap

  • Inhibition of Rho Kinases Enhances the Degradation of Mutant Huntingtin 査読

    Peter O. Bauer, Hon Kit Wong, Fumitaka Oyama, Anand Goswami, Misako Okuno, Yoshihiro Kino, Haruko Miyazaki, Nobuyuki Nukina

    JOURNAL OF BIOLOGICAL CHEMISTRY   284 ( 19 )   13153 - 13164   2009年5月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Huntington disease (HD) is a fatal hereditary neurodegenerative disease caused by an expansion of the polyglutamine (polyQ) stretch in huntingtin (htt). Whereas the pathological significance of the expanded polyQ has been clearly established and a tremendous effort to develop therapeutic tools for HD has been exerted, there is yet no effective cure. Whereas many molecules able to reduce the polyQ accumulation and aggregation have been identified, including several Rho kinase (ROCK) inhibitors, it remains very important to determine the mechanism of action of the potential drugs. ROCK inhibitors, including Y-27632 were reported to decrease aggregation of htt and androgen receptor (AR) through ROCK1 and protein kinase C-related protein kinase-2 (PRK-2). A downstream effector of ROCK1, actin-binding factor profilin, was shown to inhibit the mutant htt aggregation but not AR by direct interaction. We found that the anti-aggregation effect of ROCK inhibitors was not limited to the mutant htt and AR and that Y-27632 was also able to reduce the aggregation of ataxin-3 and atrophin-1 with expanded polyQ. These results suggested that in addition to the mechanism reported for htt and AR, there might also be other common mediators involved in the reduced aggregation of different polyQ proteins. In this study, we show that Y-27632 not only reduced the mutant htt aggregation by enhancing its degradation, but surprisingly was able to activate the main cellular degradation pathways, proteasome, and macroautophagy. We also show that this unique effect was mediated by ROCK1 and ROCK2.

    DOI: 10.1074/jbc.M809229200

    Web of Science

    PubMed

    researchmap

  • BIG-2 mediates olfactory axon convergence to target glomeruli. 査読 国際誌

    Tomomi Kaneko-Goto, Sei-Ichi Yoshihara, Haruko Miyazaki, Yoshihiro Yoshihara

    Neuron   57 ( 6 )   834 - 46   2008年3月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CELL PRESS  

    Olfactory sensory neurons expressing a given odorant receptor converge axons onto a few topographically fixed glomeruli in the olfactory bulb, leading to establishment of the odor map. Here, we report that BIG-2/contactin-4, an axonal glycoprotein belonging to the immunoglobulin superfamily, is expressed in a subpopulation of mouse olfactory sensory neurons. A mosaic pattern of glomerular arrangement is observed with strongly BIG-2-positive, weakly positive, and negative axon terminals in the olfactory bulb, which is overlapping but not identical with those of Kirrel2 and ephrin-A5. There is a close correlation between the BIG-2 expression level and the odorant receptor choice in individual sensory neurons. In BIG-2-deficient mice, olfactory sensory neurons expressing a given odorant receptor frequently innervate multiple glomeruli at ectopic locations. These results suggest that BIG-2 is one of the axon guidance molecules crucial for the formation and maintenance of functional odor map in the olfactory bulb.

    DOI: 10.1016/j.neuron.2008.01.023

    Web of Science

    PubMed

    researchmap

  • Mutant Huntingtin reduces HSP70 expression through the sequestration of NF-Y transcription factor 査読

    Tomoyuki Yamanaka, Haruko Miyazaki, Fumitaka Oyama, Masaru Kurosawa, Chika Washizu, Hiroshi Doi, Nobuyuki Nukina

    EMBO JOURNAL   27 ( 6 )   827 - 839   2008年3月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    In Huntington's disease (HD), mutant Huntingtin, which contains expanded polyglutamine stretches, forms nuclear aggregates in neurons. The interactions of several transcriptional factors with mutant Huntingtin, as well as altered expression of many genes in HD models, imply the involvement of transcriptional dysregulation in the HD pathological process. The precise mechanism remains obscure, however. Here, we show that mutant Huntingtin aggregates interact with the components of the NF-Y transcriptional factor in vitro and in HD model mouse brain. An electrophoretic mobility shift assay using HD model mouse brain lysates showed reduction in NF-Y binding to the promoter region of HSP70, one of the NF-Y targets. RT-PCR analysis revealed reduced HSP70 expression in these brains. We further clarified the importance of NF-Y for HSP70 transcription in cultured neurons. These data indicate that mutant Huntingtin sequesters NF-Y, leading to the reduction of HSP70 gene expression in HD model mice brain. Because suppressive roles of HSP70 on the HD pathological process have been shown in several HD models, NF-Y could be an important target of mutant Huntingtin.

    DOI: 10.1038/emboj.2008.23

    Web of Science

    PubMed

    researchmap

  • RNA-binding protein TLS is a major nuclear aggregate-interacting protein in huntingtin exon 1 with expanded polyglutamine-expressing cells 査読

    Hiroshi Doi, Kazumasa Okamura, Peter O. Bauer, Yoshiaki Furukawa, Hideaki Shimizu, Masaru Kurosawa, Yoko Machida, Haruko Miyazaki, Kenichi Mitsui, Yoshiyuki Kuroiwa, Nobuyuki Nukina

    JOURNAL OF BIOLOGICAL CHEMISTRY   283 ( 10 )   6489 - 6500   2008年3月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Formation of intracellular aggregates is the hallmark of polyglutamine (polyQ) diseases. We analyzed the components of purified nuclear polyQ aggregates by mass spectrometry. As a result, we found that the RNA-binding protein translocated in liposarcoma (TLS) was one of the major components of nuclear polyQ aggregate-interacting proteins in a Huntington disease cell model and was also associated with neuronal intranuclear inclusions of R6/2 mice. In vitro study revealed that TLS could directly bind to truncated N-terminal huntingtin (tNhtt) aggregates but could not bind to monomer GST-tNhtt with 18, 42, or 62Q, indicating that the tNhtt protein acquired the ability to sequester TLS after forming aggregates. Thioflavin T assay and electron microscopic study further supported the idea that TLS bound to tNhtt-42Q aggregates at the early stage of tNhtt-42Q amyloid formation. Immunohistochemistry showed that TLS was associated with neuronal intranuclear inclusions of Huntington disease human brain. Because TLS has a variety of functional roles, the sequestration of TLS to polyQ aggregates may play a role in diverse pathological changes in the brains of patients with polyQ diseases.

    DOI: 10.1074/jbc.M705306200

    Web of Science

    PubMed

    researchmap

  • BACE1 modulates filopodia-like protrusions induced by sodium channel beta 4 subunit 査読

    Haruko Miyazaki, Fumitaka Oyama, Hon-Kit Wong, Kumi Kaneko, Takashi Sakurai, Akira Tamaoka, Nobuyuki Nukina

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   361 ( 1 )   43 - 48   2007年9月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Processing of APP by BACE1 plays a crucial role in the pathogenesis of Alzheimer disease (AD). Recently, the voltage-gated sodium channel (Na-v) beta(4) subunit (beta(4)), an auxiliary subunit of Nav that is supposed to serve as a cell adhesion molecule, has been identified as a substrate for BACE1. However, the biological consequence of BACE1 processing of beta 4 remains illusive. Here, we report the biological effects of beta(4) processing by BACE1 Overexpression of beta 4 in Neuro2a cells promoted neurite extension and increased the number of F-actin rich filopodia-like protrusions. While coexpression of BACE1 together with beta(4) further accelerated neurite extension, the number of filopodia-like protrusions was reduced. Overexpression of C-terminal fragment of beta(4) that was generated by BACE1 (beta(4)-CTF) partially recapitulated the results obtained with BACE1 overexpression. These results suggest that the processing of beta(4) by BACE1 regulates neurite length and filopodia-like protrusion density in neurons. (c) 2007 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2007.06.170

    Web of Science

    PubMed

    researchmap

  • Floxed allele for conditional inactivation of the voltage-gated sodium channel beta 1 subunit Scn1b 査読

    Chunling Chen, Travis L. Dickendesher, Fumitaka Oyama, Haruko Miyazaki, Nobuyuki Nukina, Lori L. Isom

    GENESIS   45 ( 9 )   547 - 553   2007年9月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    The voltage-gated sodium channel gene Scn1b encodes the auxiliary subunit beta 1, which is widely distributed in neurons and glia of the central and peripheral nervous systems, cardiac myocytes, skeletal muscle myocytes, and neuroendocrine cells. We showed previously that the Scn1b null mutation results in a complex and severe phenotype that includes retarded growth, seizures, ataxia, and death by postnatal day 21. We generated a floxed allele of Scn1b by inserting loxP sites surrounding the second coding exon. Ubiquitous deletion of the floxed exon by Cre recombinase using CMV-Cre-transgenic mice produced the Scn1b(del) allele. The null phenotype of Scn1b(del) homozygotes is indistinguishable from that of Scn1b nulls and confirms the in vivo inactivation of Scn1b. Conditional inactivation of the floxed allele will make it possible to circumvent the lethality that results from complete loss of this gene, such that the physiological role of Scn1b in specific cell types and/or specific developmental time points can be investigated.

    DOI: 10.1002/dvg.20324

    Web of Science

    PubMed

    researchmap

  • Sodium channel beta 4 subunit: down-regulation and possible involvement in neuritic degeneration in Huntington's disease transgenic mice 査読

    F Oyama, H Miyazaki, N Sakamoto, C Becquet, Y Machida, K Kaneko, C Uchikawa, T Suzuki, M Kurosawa, T Ikeda, A Tamaoka, T Sakurai, N Nukina

    JOURNAL OF NEUROCHEMISTRY   98 ( 2 )   518 - 529   2006年7月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    Sodium channel beta 4 is a very recently identified auxiliary subunit of the voltage-gated sodium channels. To find the primarily affected gene in Huntington's disease (HD) pathogenesis, we profiled HD transgenic mice using a high-density oligonucleotide array and identified beta 4 as an expressed sequence tag (EST) that was significantly down-regulated in the striatum of HD model mice and patients. Reduction in beta 4 started at a presymptomatic stage in HD mice, whereas other voltage-gated ion channel subunits were decreased later. In contrast, spinal cord neurons, which generate only negligible levels of expanded polyglutamine aggregates, maintained normal levels of beta 4 expression even at the symptomatic stage. Overexpression of beta 4 induced neurite outgrowth in Neuro2a cells, and caused a thickening of dendrites and increased density of dendritic spines in hippocampal primary neurons, indicating that beta 4 modulates neurite outgrowth activities. These results suggest that down-regulation of beta 4 may lead to abnormalities of sodium channel and neurite degeneration in the striatum of HD transgenic mice and patients with HD.

    DOI: 10.1111/j.1471-4159.2006.03893.x

    Web of Science

    PubMed

    researchmap

  • Expanded polyglutamines impair synaptic transmission and ubiquitin-proteasome system in Caenorhabditis elegans 査読

    LA Khan, PO Bauer, H Miyazaki, KS Lindenberg, BG Landwehrmeyer, N Nukina

    JOURNAL OF NEUROCHEMISTRY   98 ( 2 )   576 - 587   2006年7月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    Polyglutamine (polyQ) expansion in many proteins, including huntingtin and ataxin-3, is pathogenic and responsible for neuronal dysfunction and degeneration. Although at least nine neurodegenerative diseases are caused by expanded polyQ, the pathogenesis of these diseases is still not well understood. In the present study, we used Caenorhabditis elegans to study the molecular mechanism of polyQ-mediated toxicity. We expressed full-length and truncated ataxin-3 with different lengths of polyQ in the nervous system of C. elegans. We show that expanded polyQ interrupts synaptic transmission, and induces swelling and aberrant branching of neuronal processes. Using an ubiquitinated fluorescence reporter construct, we also showed that polyQ aggregates impair the ubiquitin-proteasome system in C. elegans. These results may provide information for further understanding the pathogenesis of polyQ diseases.

    DOI: 10.1111/j.1471-4159.2006.03895.x

    Web of Science

    PubMed

    researchmap

  • beta subunits of voltage-gated sodium channels are novel substrates of beta-site amyloid precursor protein-cleaving enzyme (BACE1) and gamma-secretase 査読

    HK Wong, T Sakurai, F Oyama, K Kaneko, K Wada, H Miyazaki, M Kurosawa, B De Strooper, P Saftig, N Nukina

    JOURNAL OF BIOLOGICAL CHEMISTRY   280 ( 24 )   23009 - 23017   2005年6月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Sequential processing of amyloid precursor protein (APP) by membrane-bound proteases, BACE1 and gamma-secretase, plays a crucial role in the pathogenesis of Alzheimer disease. Much has been discovered on the properties of these proteases; however, regulatory mechanisms of enzyme-substrate interaction in neurons and their involvement in pathological changes are still not fully understood. It is mainly because of the membrane-associated cleavage of these proteases and the lack of information on new substrates processed in a similar way to APP. Here, using RNA interference-mediated BACE1 knockdown, mouse embryonic fibroblasts that are deficient in either BACE1 or presenilins, and BACE1-deficient mouse brain, we show clear evidence that beta subunits of voltage-gated sodium channels are sequentially processed by BACE1 and gamma-secretase. These results may provide new insights into the underlying pathology of Alzheimer disease.

    DOI: 10.1074/jbc.M414648200

    Web of Science

    PubMed

    researchmap

  • Decreased expression of hypothalamic neuropeptides in Huntington disease transgenic mice with expanded polyglutamine-EGFP fluorescent aggregates 査読

    S Kotliarova, NR Jana, N Sakamoto, M Kurosawa, H Miyazaki, M Nekooki, H Doi, Y Machida, HK Wong, T Suzuki, C Uchikawa, Y Kotliarov, K Uchida, Y Nagao, U Nagaoka, A Tamaoka, K Oyanagi, F Oyama, N Nukina

    JOURNAL OF NEUROCHEMISTRY   93 ( 3 )   641 - 653   2005年5月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING LTD  

    Huntington disease is caused by polyglutamine (polyQ) expansion in huntingtin. Selective and progressive neuronal loss is observed in the striatum and cerebral cortex in Huntington disease. We have addressed whether expanded polyQ aggregates appear in regions of the brain apart from the striatum and cortex and whether there is a correlation between expanded polyQ aggregate formation and dysregulated transcription. We generated transgenic mouse lines expressing mutant truncated N-terminal huntingtin (expanded polyQ) fused with enhanced green fluorescent protein (EGFP) and carried out a high-density oligonucleotide array analysis using mRNA extracted from the cerebrum, followed by TaqMan RT-PCR and in situ hybridization. The transgenic mice formed expanded polyQ-EGFP fluorescent aggregates and this system allowed us to directly visualize expanded polyQ aggregates in various regions of the brain without performing immunohistochemical studies. We show here that polyQ-EGFP aggregates were intense in the hypothalamus, where the expression of six hypothalamic neuropeptide mRNAs, such as oxytocin, vasopressin and cocaine-amphetamine-regulated transcript, was down-regulated in the transgenic mouse brain without observing a significant loss of hypothalamic neurons. These results indicate that the hypothalamus is susceptible to aggregate formation in these mice and this may result in the down-regulation of specific genes in this region of the brain.

    DOI: 10.1111/j.1471-4159.2005.03035.x

    Web of Science

    PubMed

    researchmap

  • Gem GTPase and Tau - Morphological changes induced by Gem GTPase in Cho cells are antagonized by tau 査読

    F Oyama, S Kotliarova, A Harada, M Ito, H Miyazaki, Y Ueyama, N Hirokawa, N Nukina, Y Ihara

    JOURNAL OF BIOLOGICAL CHEMISTRY   279 ( 26 )   27272 - 27277   2004年6月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    A series of observations have indicated that tau, one of the major microtubule-associated proteins, is involved in neuronal cell morphogenesis and axonal maintenance. Tau is also the major component of paired helical filaments found in brains affected by Alzheimer's disease. To explore an as yet unidentified role of tau in vivo, similar to 11,000 mRNAs were profiled from tau-deficient mouse brains and compared with those from control brains at the same ages. The expression of Gem GTPase, a small GTP-binding protein of the ras superfamily, was significantly increased in the brains of tau-deficient mice at 8 weeks of age. Because Gem GTPase is a negative regulator of the Rho-Rho kinase pathway for cytoskeletal organization, this protein was transiently overexpressed in Chinese hamster ovary cells that do not express tau. Overexpression of Gem GTPase induced a marked elongation of Chinese hamster ovary cells, and simultaneous expression of tau eliminated this effect, although tau did not bind directly to Gem GTPase. This anti-elongation activity of tau was attributed to its microtubule-binding domain, and homologous domains of microtubule-associated proteins 2 and 4 exhibited similar antagonistic activities. Taken together, the present results indicate that the level of Gem GTPase and its cell elongation activity are modulated by tau and suggest that tau may be involved in a Gem GTPase-mediated signal transduction pathway.

    DOI: 10.1074/jbc.M401634200

    Web of Science

    PubMed

    researchmap

  • Pro-apoptotic protein kinase C delta is associated with intranuclear inclusions in a transgenic model of Huntington's disease 査読

    EA Zemskov, NR Jana, M Kurosawa, H Miyazaki, N Sakamoto, M Nekooki, N Nukina

    JOURNAL OF NEUROCHEMISTRY   87 ( 2 )   395 - 406   2003年10月

     詳細を見る

    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING LTD  

    In order to investigate any effect of truncated mutant huntingtin (tNhtt) aggregation on protein kinase C (PKC) signaling in Huntington's disease (HD), we studied a possible association of PKC isoforms with the aggregates using cellular and transgenic models of HD. In this report we describe an association of mutant tNhtt with at least three PKC isoforms (alpha, delta, zeta), as revealed by co-immunoprecipitation assays and immunocytochemistry in a cellular model of HD (Neuro2a cells expressing tNhtt-150Q-EGFP), as well as a specific association of PKCdelta with intranuclear aggregates in a transgenic model (R6/2 mice). Immunoblot analysis of isolated nuclear fractions shows an elevation of nuclear PKCdelta in transgenic brain tissue. The observed elevation has a strong similarity with the apoptotic translocation of PKCdelta detected in experiments with the mouse neuroblastoma Neuro2a cells. Using a Neuro2a cell line expressing tNhtt with the nuclear localization signal, we demonstrate the association of PKCdelta with intranuclear aggregates and present evidence that accumulation of PKCdelta in cell nuclei does not depend on mutant htt nuclear translocation. Our results suggest that the association of PKCdelta with intranuclear htt-aggregates may affect its apoptotic function in a transgenic model of HD.

    DOI: 10.1046/j.1471-4159.2003.02002.x

    Web of Science

    PubMed

    researchmap

▼全件表示

MISC

  • マウス脳における脳梁離断を用いた線維状αsynucleinの伝播経路の検討

    奥住 文美, 波田野 琢, 黒澤 大, 山中 智行, 宮崎 晴子, 古川 良明, 服部 信孝, 貫名 信行

    パーキンソン病・運動障害疾患コングレスプログラム・抄録集   11回   68 - 68   2017年10月

     詳細を見る

    記述言語:日本語   出版者・発行元:Movement Disorder Society of Japan (MDSJ)  

    researchmap

  • 電位依存性ナトリウムチャネルβサブユニットとてんかん関連変異の分子構造と機能解析

    清水英明, 宮崎晴子, 庄司志咲子, 白水美香子, 貫名信行, 横山茂之

    第35回日本神経科学大会   P4-j19   2012年9月

     詳細を見る

    記述言語:日本語   掲載種別:研究発表ペーパー・要旨(全国大会,その他学術会議)  

    researchmap

  • 電位依存性ナトリウムチャネルβサブユニットの分子間相互作用の解析

    清水英明, 宮崎晴子, 庄司志咲子, 白水美香子, 貫名信行, 横山茂之

    日本分子生物学会年会プログラム・要旨集(Web)   34th   4P-0276   2011年12月

     詳細を見る

    記述言語:日本語   掲載種別:研究発表ペーパー・要旨(全国大会,その他学術会議)  

    J-GLOBAL

    researchmap

  • Correlation between nuclear accumulation and dysregulation of sodium channel beta 4 subunit in HD transgenic mice

    Haruko Miyazaki, Fumitaka Oyama, Masaru Kurosawa, Mizuki Yamada, Nobuyuki Nukina

    NEUROSCIENCE RESEARCH   65   S247 - S247   2009年

     詳細を見る

    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    DOI: 10.1016/j.neures.2009.09.1400

    Web of Science

    researchmap

  • Scn4b Is A Genetic Modifier Of Cardiac Conduction Disease In Mice

    Brandon P. Scicluna, Carol A. Remme, Arie O. Verkerk, Ahmad S. Amin, Michael W. Tanck, Leander Beekman, Vera H. Deneer, Catherine Chevaller, Fumitaka Oyama, Haruko Miyazaki, Nobuyuki Nukina, Denis Escande, Remi Houlgatte, Jacques M. de Bakker, Marieke Veldkamp, Hanno L. Tan, Arthur A. Wilde, Connie R. Bezzina

    CIRCULATION   118 ( 18 )   S339 - S339   2008年10月

     詳細を見る

    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:LIPPINCOTT WILLIAMS & WILKINS  

    Web of Science

    researchmap

  • Mutant Huntingtin reduces HSP70 expression through the sequestration of NF-Y transcription factor

    Tomoyuki Yamanaka, Haruko Miyazaki, Fumitaka Oyama, Masaru Kurosawa, Chika Washizu, Hiroshi Doi, Nobuyuki Nukina

    NEUROSCIENCE RESEARCH   61   S45 - S45   2008年

     詳細を見る

    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    Web of Science

    researchmap

  • Downregulation of sodium channel beta 4 in Huntington disease transgenic mice

    Fumitaka Oyama, Haruko Miyazaki, Masaru Kurosavva, Mizuki Yamada, Nobuyuki Nukina

    NEUROSCIENCE RESEARCH   61   S204 - S204   2008年

     詳細を見る

    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    Web of Science

    researchmap

  • beta subunits of voltage-gated sodium channels are novel substrates of beta-site amyloid precursor protein-cleaving enzyme (BACE1) and gamma-secretase

    T Sakurai, HK Wong, F Oyama, K Kaneko, K Wada, H Miyazaki, M Kurosawa, B De Strooper, P Saftig, N Nukina

    JOURNAL OF PHARMACOLOGICAL SCIENCES   100   244P - 244P   2006年

     詳細を見る

    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:JAPANESE PHARMACOLOGICAL SOC  

    Web of Science

    researchmap

  • Dysregulation of sodium channel beta 4 subunit by expanded polyglutamine in Huntington disease transgenic mice

    Fumitaka Oyama, Haruko Miyazaki, Kazumasa Okamura, Yoko Machida, Kurosawa Masaru, Takashi Sakurai, Nobuyuki Nukina

    NEUROSCIENCE RESEARCH   55   S257 - S257   2006年

     詳細を見る

    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    Web of Science

    researchmap

  • Region specific changes of gene expression in a transgenic mouse model for Huntington disease.

    F Oyama, H Miyazaki, N Sakamoto, NR Jana, SE Kotliarova, N Nukina

    AMERICAN JOURNAL OF HUMAN GENETICS   73 ( 5 )   544 - 544   2003年11月

     詳細を見る

    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:UNIV CHICAGO PRESS  

    Web of Science

    researchmap

▼全件表示

講演・口頭発表等

  • 大脳皮質-黒質神経回路のトレーシングおよび免疫組織化学的解析

    宮﨑 晴子, 立川 哲也, 平井 康治, 苅部 冬紀, 宮坂 知宏, 藤山 文乃, 山川 和弘, 貫名 信行

    第44回 日本神経科学大会  2021年7月31日 

     詳細を見る

    開催年月日: 2021年7月28日 - 2021年7月31日

    記述言語:英語   会議種別:ポスター発表  

    添付ファイル: 4P-161_宮﨑.pdf

    researchmap

  • 黒質に投射する新規中枢無髄神経回路の探索

    宮崎 晴子, 立川 哲也, 山川 和弘, 貫名 信行

    第43回 日本神経科学大会  2020年7月31日 

     詳細を見る

    開催年月日: 2020年7月29日 - 2020年8月1日

    記述言語:英語   会議種別:ポスター発表  

    researchmap

受賞

  • 大藤内分泌医学賞

    2021年8月   令和3年度 大藤内分泌医学賞   ハンチントン病モデルマウスにおける神経ペプチド遺伝子群発現低下メカニズムの解析

     詳細を見る

  • 学術奨励賞

    2007年8月   病態と治療におけるプロテアーゼとインヒビター学会  

    宮崎 晴子

     詳細を見る

共同研究・競争的資金等の研究

  • 中枢無髄神経における自閉症スペクトラム障害関連蛋白質Nav1.2の分布の検討

    2020年10月 - 2021年10月

    公益財団法人金原一郎記念医学医療振興財団  第35回基礎医学医療研究助成金 

    宮﨑 晴子

      詳細を見る

    担当区分:研究代表者 

    researchmap

  • 中枢無髄神経特異的に分布する蛋白質の探索

    2018年10月 - 2021年05月

    公益財団法人 武田科学振興財団  ライフサイエンス研究助成 

    宮崎 晴子

      詳細を見る

    担当区分:研究代表者  資金種別:競争的資金

    researchmap

  • 髄鞘化における有髄・無髄の決定にかかわるメカニズムの解析

    2018年04月 - 2019年03月

    同志社大学赤ちゃん学研究センター  計画共同研究  計画共同研究

      詳細を見る

    担当区分:研究代表者 

    researchmap

  • 中枢神経系無髄神経の包括的分子機能解析

    研究課題/領域番号:17H01564  2017年04月 - 2022年03月

    日本学術振興会  科学研究費助成事業 基盤研究(A)  基盤研究(A)

    貫名 信行, 山川 和弘, 宮崎 晴子

      詳細を見る

    配分額:41860000円 ( 直接経費:32200000円 、 間接経費:9660000円 )

    中枢神経系の無髄神経の分布に関してNav1.2をマーカーにして検討を行っている。脳梁・分界条の線維に無髄が存在することを免疫電顕を含めて確認した。現在その起始細胞を同定しようと検討している。
    多系統萎縮症において線条体のNav1.2の染色性の脱落を見出した。この起始核である中型有棘細胞の脱落との関係を検討中である。無髄関連タンパク質と考えているGolliについてはconditional knock outマウス作成のためのFloxマウスの作製に成功した。現在その確認を行っている。Nav1.2のconditional knockoutマウスについては線条体、小脳顆粒細胞におけるknockoutを、作製中である。
    無髄神経関連タンパク質のプロテオーム解析は有髄線維の混入のため、難航している。線条体中型有棘細胞を蛍光ラベルしたマウスの使用なども考慮中である。ハンチントン病中型有棘細胞のtranscriptome解析を行い、データを投稿中である。この投射線維は無髄であり、プロテオーム解析と合わせて、解析を進められる可能性が出てきた。
    線条体の回路は皮質線条体とサーキットを作っているが、皮質線条体がStxbp1やScn2aのヘテロ欠損マウスにおけるてんかんの発症に関与していることを示した(Miyamoto et al Nat Commun2019,Tatsukawa et al Mol Autism, Ogiwara et al Commun Biol2018)。

    researchmap

  • 中枢無髄神経の機能解析に関する研究基盤の確立

    研究課題/領域番号:16K07005  2016年04月 - 2020年03月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    宮崎 晴子, 下郡 智美

      詳細を見る

    配分額:4940000円 ( 直接経費:3800000円 、 間接経費:1140000円 )

    中枢神経系には海馬の苔状線維、小脳の平行線維、線条体投射神経線維など意外にも多くの無髄神経が保存されているが、それらの役割についてはほとんどわかっていない。本研究では中枢無髄神経の役割を解明するための研究基盤を確立すべく、無髄神経特異的に分布する分子の同定、および無髄神経特異的な蛋白質複合体を同定することを目的としている。今年度は中枢の無髄神経のひとつである線条体投射神経を用いて、無髄線維に存在する蛋白質の網羅的解析を行った。
    8-12週齢のマウス脳から線条体投射神経線維を多く含む領域を採取し、超遠心を行って膜画分を分離した後スクロース密度勾配遠心によりミエリン(+)、ミエリン(-)画分に分離した。続いてwestern blotを行い、ミエリン(-)画分ではミエリンの主要蛋白質であるMBP(myelin basic protein)がほとんど除かれていることを確認した。これらの画分に存在する蛋白質をSDS-PAGEで比較したところ、差のあるバンドが多数得られたことから、ミエリン(-)画分には無髄神経特有の蛋白質が含まれている可能性が考えられた。続いてそれぞれの画分についてLC-MS/MS解析を行い、MascotScoreのratioがミエリン(-)/ミエリン(+)>1.5となるような蛋白質を「ミエリン(-)画分に濃縮して存在する蛋白質」としてデータから抽出した。このようにして得られた蛋白質にはミエリンインヒビターとして知られているMAG (myelin associated glycoprotein)のレセプターが含まれていたことから、下流のmTORを介したシグナルカスケードが中枢無髄線維の「非髄鞘化」メカニズムに関与している可能性が示された。

    researchmap

  • ポリグルタミン病における長鎖ncRNA異常の解析

    研究課題/領域番号:25253066  2013年04月 - 2016年03月

    日本学術振興会  科学研究費助成事業 基盤研究(A)  基盤研究(A)

    貫名 信行, 宮﨑 晴子, 下郡 智美

      詳細を見る

    配分額:45110000円 ( 直接経費:34700000円 、 間接経費:10410000円 )

    ハンチントン病(HD)における遺伝子発現異常と病態との関連を明らかにするために線条体中型有棘細胞(MSN)特異的な遺伝子発現を検討する方法を確立した。セルソーターを用いて蛍光蛋白質を発現したMSNを対照とHDモデルマウスとから精製し、この遺伝子発現をマイクロアレーを用いて検討した。MSN特異的遺伝子発現が 検出されているかどうかをRT-PCRによって検討し、発現量がある程度以上であれば信頼性のある結果がマイクロアレーで得られていることを確認した。long non-coding RNAの異常もこの方法によって同定できたが、一般発現量が少ないことが多いため、確認には新たな方法の確立が必要である。

    researchmap

  • 選択的ニューロン病態解析法の開発・展開

    研究課題/領域番号:22110004  2010年04月 - 2015年03月

    日本学術振興会  科学研究費助成事業 新学術領域研究(研究領域提案型)  新学術領域研究(研究領域提案型)

    貫名 信行, 宮﨑 晴子, 山中 智行, 松本 弦

      詳細を見る

    配分額:111670000円 ( 直接経費:85900000円 、 間接経費:25770000円 )

    ハンチントン病における選択的神経細胞変性の機序を明らかにするため、凝集体結合タンパク質FUS/TLS,NF-YA,p62のノックアウトマウスも含めた解析を行い、それぞれのin vivoの機能を明らかにするとともに、ポリグルタミン病病態との関連を検討した。遺伝子発現異常を呈するsodium channel beta4 subunitについて同様にノックアウトマウスの解析から線条体中型有棘神経細胞でリサージェントカレントを制御していることを示した。またその分布から大脳基底核投射線維が無髄であるという新しい解剖学的事実を提示した。またセルソーターを用いた新しい細胞機能解析手法を確立した。

    researchmap

▼全件表示