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Hiroki Shibata Last modified date:2023.06.27

Associate Professor / Division of Medical Molecular Cell Biology, Graduate School of Systems Life Sciences
Medical Research Center for High Depth Omics
Medical Institute of Bioregulation


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Homepage
https://kyushu-u.pure.elsevier.com/en/persons/hiroki-shibata
 Reseacher Profiling Tool Kyushu University Pure
http://gen.kyushu-u.ac.jp/~byouin/
Introduction of research projects, publications and members of the laboratory. .
Phone
092-642-6168
Fax
092-632-2375
Academic Degree
Ph D.
Country of degree conferring institution (Overseas)
No
Field of Specialization
Genetic, Genomics, Population Genetics
Total Priod of education and research career in the foreign country
03years01months
Outline Activities
Two major research projects (both heavily using NGS) are ongoing:
1. Exome sequencing is a powerful technique to identify causative variants of genetic diseases (Figure). However, the filtration of numerous candidate variants can be very difficult especially diseases so rare that a single pedigree ascertained. We introduced SNP linkage information for the filtration (Linkage-assisted exome sequencing). We are currently applying the technique on the analyses of multiple hereditary neurological diseases, such as idiopathic hyper CK-emia, spastic paraplegia and familial myochronic epilepsy.
2. Venoms are promising seeds for pharmaceutical discovery. Japanese pit viper, Habu snake (Protobothrops flavoviridis) is well known for the dangerous bites. However most of the venomic proteins are uncharacterized. Also accelerated evolution has been reported in many of the venomic proteins. Towards the complete characterization of the venomic proteins and the understanding of the mechanism of their accelerated evolution, we are currently working on omics studies of Habu snake, including whole genome sequencing, RNA-sequencing and population genetics studies. We are also participating in international research projects of whole genome sequencing of Elapidae (cobras).
Research
Research Interests
  • Genetic divergence and diversity of venom proteins among island populations of Japanese Habu snakes, Protobothrops.
    keyword : Protobothrops flavoviridis, Protobothrops tokarensis, Protopbothrops elegans, venom protein gene, genetic divergence, genetic diversity,
    2012.04.
  • Ddhd1 KO mouse as a model of spastic paraplegia
    keyword : spastic paraplegia, Ddhd1, CRISPR, RNA-seq, metabolome analysis
    2015.04.
  • Divergence and diversity of mitogenome among island populations of Japanese Habu snakes, Protobothrops.
    keyword : Protobothrops flavoviridis, Protobothrops tokarensis, Protopbothrops elegans, mitochondria genome, microsatellite marker, genetic divergence, genetic diversity,
    2012.04.
  • Construction of the full length gene catalogue of Japanese Habu viper, Protobothrops flavoviridis.
    keyword : Protobothrops flavoviridis, next generation sequencer, RNA-sequencing, cDNA, venom gland, snake venoms
    2011.09.
  • Whole genome sequencing of Japanese Habu viper, Protobothrops flavoviridis.
    keyword : Protobothrops flavoviridis, next generation sequencer, denovo assembly, scaffolding, gene prediction, snake venoms
    2012.04.
  • Genetic analysis of spastic paraplegia
    keyword : spinocerebellar ataxia, linkage analysis, next generation sequencer, exome sequencing
    2014.04.
  • Genetic analysis of spinocerebellar ataxia
    keyword : spinocerebellar ataxia, linkage analysis, next generation sequencer, exome sequencing
    2014.04.
  • Genetic analysis of familial myoclonic epilepsy
    keyword : familial myoclonic epilepsy, linkage analysis, next generation sequencer, exome sequencing
    2011.04.
  • Exome sequencing approach to identify the responsible variant for a hereditary motor and sensory neuropathy
    keyword : Hereditary motor and sensory neuropathy, linkage analysis, next generation sequencer, exome sequencing
    2010.04.
  • Evolutionary study of schizophrenia-associated genes.
    keyword : schizophrenia, glutamate receptor, molecular evolution, genetic diversity, natural selection
    2004.04~2013.03.
Academic Activities
Books
1. Tomohisa Ogawa, Hiroki Shibata, Venomics study of Protobothrops flavoviridis snake: Habu snake venomics: How venom proteins have evolved?, IntechOpen, 2020.04, Venomics projects have been conducted to disclose the divergent profiles and evolution of various venomous animals. Here, we describe the venomics project including genome and transcriptome of Habu snake leading to drug discovery..
Papers
1. Isomoto A, Shoguchi E, Hisata K, Inoue J, Sun Y, Inaba K, Satoh N, Ogawa T, Shibata H., Active expression of genes for protein modification enzymes in the venom gland of habu snakes, Toxins (Basel), 14, 300, 2022.04, Genes encoding snake venom toxins have been studied extensively. However, genes involved in the modification and functioning of venom proteins are little known. Protobothrops is a genus of pit vipers, which are venomous and inhabit the Nansei (Southwest) islands of Japan, Taiwan China, Vietnam, Thailand, Myanmar, Nepal, Bhutan, and India. Our previous study decoded the genome of Protobothrops flavoviridis, a species endemic to the Nansei Islands, Japan, and revealed unique evolutionary processes of some venom genes. In this study, we analyzed genes that are highly expressed in venom glands to survey genes for candidate enzymes or chaperone proteins involved in toxin folding and modification. We found that, in addition to genes that encode venom proteins and ribosomal proteins, genes that encode protein disulfide isomerase (PDI) family members (orthologs of human P4HB and PDIA3), Selenoprotein M (SELENOM), and Calreticulin (CALR) are highly expressed in venom glands. Since these enzymes or chaperones are involved in protein modification and potentially possess protein folding functions, we propose that P4HB, SELENOM, CALR, and PDIA3 encode candidate enzymes or chaperones to confer toxic functions upon the venom transcriptome..
2. Miura S, Shimojo T, Morikawa T, Kamada T, Uchiyama Y, Kurata S, Fujioka R, Shibata H., Familial paroxysmal kinesigenic dyskinesia with a novel missense variant (Arg2866Trp) in NBEA., J Hum Genet., 66, 8, 805-811, 2021.03, Paroxysmal kinesigenic dyskinesia (PKD) is a movement disorder characterized by episodic involuntary movement attacks triggered by sudden movements, acceleration, or intention to move. We ascertained two Japanese familial cases with PKD. The proband is a 22-year-old woman who had noted sudden brief ( C [p.Gln2992His] in FAT2 and NM_015678: c.8596C > T [p.Arg2866Trp] in NBEA). Real time quantitative PCR analysis of Nbea mRNA levels in the developing rat brain revealed peak at postnatal day 28 and decline at postnatal day 56. This result might match the most common clinical course of PKD from the point of view of the most common age at remission. NBEA has been reported to be responsible for neurodevelopmental disease accompanied by epilepsy. We concluded the variant in NBEA most likely to be responsible for our familial cases of PKD..
3. Morikawa T, Ohishi H, Kosaka K, Shimojo T, Nagano A, Taniguchi I, Fujioka R, Moriyama K, Unoki M, Takahashi M, Nakao M, Izumi Y, Bamba T, Sasaki H, Miura S, Shibata H. , Ddhd1 knockout mouse as a model for familial spastic paraplegia, Biosci Rep., 41, 2, BSR20204171, 2021.02, We have previously reported a novel homozygous 4-bp deletion in DDHD1 as the respon- sible variant for spastic paraplegia type 28 (SPG28; OMIM#609340). The variant causes a frameshift, resulting in a functionally null allele in the patient. DDHD1 encodes phospholi- pase A1 (PLA1) catalyzing phosphatidylinositol to lysophosphatidylinositol (LPI). To clarify the pathogenic mechanism of SPG28, we established Ddhd1 knockout mice (Ddhd1[−/−]) carrying a 5-bp deletion in Ddhd1, resulting in a premature termination of translation at a position similar to that of the patient. We observed a significant decrease in foot–base angle (FBA) in aged Ddhd1(−/−) (24 months of age) and a significant decrease in LPI 20:4 (sn-2) in Ddhd1(−/−) cerebra (26 months of age). These changes in FBA were not observed in 14 months of age. We also observed significant changes of expression levels of 22 genes in the Ddhd1(−/−) cerebra (26 months of age). Gene Ontology (GO) terms relating to the nervous system and cell–cell communications were significantly enriched. We conclude that the re- duced signaling of LPI 20:4 (sn-2) by PLA1 dysfunction is responsible for the locomotive abnormality in SPG28, further suggesting that the reduction of downstream signaling such as GPR55 which is agonized by LPI is involved in the pathogenesis of SPG28..
4. Miura S, Kosaka K, Shimojo T, Matsuura E, Noda K, Fujioka R, Mori S, Umehara F, Iwaki T, Yamamoto K, Saitsu H, Shibata H., Intronic variant in IQGAP3 associated with hereditary neuropathy with proximal lower dominancy, urinary disturbance, and paroxysmal dry cough., J Hum Genet., 65, 9, 717-725, 2020.09, In 2008, we reported a clinically and genetically new type of autosomal dominant disorder of motor and sensory neuropathy with proximal dominancy in the lower extremities, urinary disturbance, and paroxysmal dry cough. To identify the nucleotide variant causative of this disease, we reanalyzed the linkage of the original Japanese pedigree including seven newly ascertained subjects with updated information. We assigned the locus of the disease to 1p13.3-q23 (maximum logarithm-of-odds score = 2.71). Exome sequencing for five patients and one healthy relative from the pedigree revealed 2526 patient-specific single- nucleotide variants (SNVs). By rigorous filtering processes using public databases, our linkage results, and functional prediction, followed by Sanger sequencing of the pedigree and 520 healthy Japanese individuals, we identified an intronic SNV in IQGAP3, a gene known to be associated with neurite outgrowth. Upon pathological examination of the sural nerve, moderate, chronic, mainly axonal neuropathy was observed. By histochemical analyses, we observed a patient-specific increase of IQGAP3 expression in the sural nerve. We concluded that the variant of IQGAP3 is associated with the disease in our pedigree..
5. Ogawa T, Oda-Ueda N, Hisata K, Nakamura H, Chijiwa T, Hattori S, Isomoto A, Yugeta H, Yamasaki S, Fukumaki Y, Ohno M, Satoh N, Shibata H., Alternative mRNA splicing in three venom families underlying a possible production of divergent venom proteins of the Habu snake, Protobothrops flavoviridis., Toxins, 11, 10, 581-581, 2019.10, Snake venoms are complex mixtures of toxic proteins encoded by various gene families that function synergistically to incapacitate prey. A huge repertoire of snake venom genes and proteins have been reported, and alternative splicing is suggested to be involved in the production of divergent gene transcripts. However, a genome-wide survey of the transcript repertoire and the extent of alternative splicing still remains to be determined. In this study, the comprehensive analysis of transcriptomes in the venom gland was achieved by using PacBio sequencing. Extensive alternative splicing was observed in three venom protein gene families, metalloproteinase (MP), serine protease (SP), and vascular endothelial growth factors (VEGF). Eleven MP and SP genes and a VEGF gene are expressed as a total of 81, 61, and 8 transcript variants, respectively. In the MP gene family, individual genes are transcribed into different classes of MPs by alternative splicing. We also observed trans-splicing among the clustered SP genes. No other venom genes as well as non-venom counterpart genes exhibited alternative splicing. Our results thus indicate a potential contribution of mRNA alternative and trans-splicing in the production of highly variable transcripts of venom genes in the habu snake..
6. Hiroki Shibata, Takahito Chijiwa, Naoko Oda-Ueda, Hitomi Nakamura, Kazuaki Yamaguchi, Shousaku Hattori, Kazumi Matsubara, Yoichi Matsuda, Akifumi Yamashita, Akiko Isomoto, Kazuki Mori, Kosuke Tashiro, Satoru Kuhara, Shinichi Yamasaki, Manabu Fujie, Hiroki Goto, Ryo Koyanagi, Takeshi Takeuchi, Yasuyuki Fukumaki, Motonori Ohno, Eiichi Shoguchi, Kanako Hisata, Noriyuki Satoh, Tomohisa Ogawa, The habu genome reveals an evolutionary background of venom protein genes., Sceintific Reports, 8: 11300., 2018.07, Evolution of novel traits is a challenging subject in biological research. Several snake lineages developed elaborate venom systems to deliver complex protein mixtures for prey capture. To understand mechanisms involved in snake venom evolution, we decoded here the ~1.4-Gb genome of a habu, Protobothrops flavoviridis. We identified 60 snake venom protein genes (SV) and 224 nonvenom paralogs (NV), belonging to 18 gene families. Molecular phylogeny reveals early divergence of SV and NV genes, suggesting that one of the four copies generated through two rounds of whole genome duplication was modified for use as a toxin. Among them, both SV and NV genes in four major components were extensively duplicated after their diversification, but accelerated evolution is evident exclusively in the SV genes. Both venom-related SV and NV genes are significantly enriched in microchromosomes. The present study thus provides a genetic background for evolution of snake venom composition..
7. Hiroki Shibata, Shuichi Sakata, Yuzo Hirano, Eiji Nitasaka, Ai Sakabe. , Facultative parthenogenesis validated by DNA analyses in the green anaconda (Eunectes murinus)., Plos ONE, 12, 12, e0189654, 2017.12, In reptiles, the mode of reproduction is typically sexual. However, facultative parthenogenesis occurs in some Squamata, such as Komodo dragon (Varanus komodoensis) and Burmese python (Python bivittatus). Here, we report facultative parthenogenesis in the green anaconda (Eunectes murinus). We found two fully developed female neonates and 17 undeveloped eggs in the oviduct of a female anaconda isolated from other individuals for eight years and two months at Ueno Zoo, Japan. To clarify the zygosity of the neonates, we analyzed 18 microsatellite markers of which 16 were informative. We observed only maternal alleles and no paternal alleles for all 16 markers. To examine the possibility of the long-term sperm storage, we estimated allele frequencies in a putative parental stock by genotyping five unrelated founders. If all founders, including the mother, are originated from a single Mendelian population, then the probability that the neonates were produced by sexual reproduction with an unrelated male via long-term sperm storage was infinitesimally small (2.31E-32 per clutch). We also examined samples from two additional offspring that the mother delivered eight years before her death. We consistently observed paternal alleles in these elder offspring, indicating that the mother had switched from sexual reproduction to asexual reproduction during the eight years of isolation. This is the first case of parthenogenesis in Eunectes to be validated by DNA analysis, and suggests that facultative parthenogenesis is widespread in the Boidae..
8. Shiroh Miura, Takuya Morikawa, Ryuta Fujioka, Kazuhito Noda, Kengo Kosaka, Takayuki Taniwaki, Hiroki Shibata, A novel missense variation (Q220R) of GNB4 encoding a guanine nucleotide-binding protein, beta-4 in a Japanese autosomal dominant motor and sensory neuropathy family., European Journal of Medical Genetics, 60, 9, 474-478, 2016.06, Dominant intermediate CharcoteMarieeTooth disease F (CMTDIF) is an autosomal dominant hereditary form of CharcoteMarieeTooth disease (CMT) caused by variations in the guanine nucleotide-binding protein, subunit beta-4 gene (GNB4). We examined two Japanese familial cases with CMT. Case 1 was a 49-year-old male whose chief complaint was slowly progressive gait disturbance and limb dysesthesia that appeared at the age of 47. On neurological examination, he showed hyporeflexia or areflexia, distal limb muscle weakness, and distal sensory impairment with lower dominancy. Nerve conduction studies demonstrated demyelinating sensorimotor neuropathy with reduced action potentials in the lower limbs. Case 2 was an 80-year-old man, Case 1's father, who reported difficulty in riding a bicycle at the age of 76. On neurological examination, he showed areflexia in the upper and lower limbs. Distal sensory
impairment in the lower limbs was also observed. Nerve conduction studies revealed mainly axonal involvement. Exome sequencing identified a novel heterozygous nonsynonymous variant (NM_021629.3:c.659T > C [p.Gln220Arg]) in GNB4 exon 8, which is known to be responsible for CMT. Sanger sequencing confirmed that both patients are heterozygous for the variation, which causes an amino acid substitution, Gln220Arg, in the highly conserved region of the WD40 domain of GNB4. The frequency of this variant in the Exome Aggregation Consortium Database was 0.000008247, and we confirmed its absence in 502 Japanese control subjects. We conclude that this novel GNB4 variant is causative for CMTDIF in these patients, who represent the first record of the disease in the Japanese
population..
9. Shiroh Miura, Takuya Morikawa, Ryuta Fujioka, Kengo Kosaka, Kohei Yamada, Kazuhito Noda, Gohsuke Hattori, Manabu Motomura, Takayuki Taniwaki, Hiroki Shibata, A novel frameshift mutation of DDHD1 in a Japanese patient with autosomal recessive spastic paraplegia., European Journal of Medical Genetics, 10.1016/j.ejmg.2016.05.010, 59, 8, 413-416, 2016.05, Spastic paraplegia (SPG) type 28 is an autosomal recessive SPG caused by mutations in the DDHD1 gene. We examined a Japanese 54-years-old male patient with autosomal recessive SPG. His parents were consanguineous. He needed a wheelchair for transfer due to spastic paraplegia. There was a history of operations for bilateral hallux valgus, thoracic ossification of the yellow ligament, bilateral carpal tunnel syndrome, bilateral ankle contracture, and lumbar spinal canal stenosis. He noticed gait disturbance at age 14. He used a cane for walking in his 40s. On neurological examination, he showed hyperreflexia, spasticity, and weakness in the lower extremities and bilateral Babinski reflexes. Urinary dysfunctions and impaired vibration sense in the lower limbs were observed. By exome sequencing analysis using Agilent SureSelect and Illumina MiSeq, we identified 17,248 homozygous nucleotide variants in the patient. Through the examination of 48 candidate genes known to be responsible for autosomal recessive SPG, we identified a novel homozygous 4-bp deletion, c.914_917delGTAA, p.Ser305Ilefs*2 in exon2 of the DDHD1 gene encoding phosphatidic acid-preferring phospholipase A1 (PA-PLA1). The mutation is expected to cause a frameshift generating a premature stop codon 3-bp downstream from the deletion. In consequence, the DDHD domain that is known to be critical for PLA1 activity is completely depleted in the mutated DDHD1 protein, predicted to be a functionally null mutation of the DDHD1 gene. By Sanger sequencing, we confirmed that both parents are heterozygous for the mutation. This variation was not detected in 474 Japanese control subjects as well as the data of the 1,000G Project. We conclude that the novel mutation in DDHD1 is the causative variant for the SPG28 patient that is the first record of the disease in Japanese population..
10. Hiroki Shibata, Takahito Chijiwa, Shosaku Hattori, Koki Terada, Motonori Ohno, Yasuyuki Fukumaki, The taxonomic position and the unexpected divergence of the Habu viper, Protobothrops among Japanese subtropical islands., Molecular Phylogenetics and Evolution, 10.1016/j.ympev.2016.04.027, 101, 91-100, 2016.04, There are four Habu species currently recognized in Japan: Protobothrops flavoviridis from the Amami Islands and the Okinawa Islands, P. tokarensis from the Tokara Islands, P. elegans from the Yaeyama Islands and Ovophis okinabvensis from the Amami Islands and the Okinawa Islands. To clarify their taxonomic positions, we determined the complete mitochondria genome sequence (approx. 17 kb) from two specimens from two different islands each for P. flavoviridis, P. tokarensis and P. elegans as well as one specimen of O. okinavensis and reconstructed the molecular phylogeny of Protobothrops using the published sequences of related species. The maximum likelihood tree showed four major species groups within Protbothrops: Group I consisting of P. cornutus, P. dabieshanensis, P. jerdonii and P. xiangchengensis; Group II consisting of P. flavoviridis and P. tokarensis; Group III consisting of P. maolensis, P. mucrosquamatus and P. elegans; Group IV consisting of P. himalayanus and P. kaubacki. Since we observed an unexpected divergence and the paraphyly of the two samples of P. flavoviridis collected from different islands, Amami-Oshima and Okinawajima within the Group II, we expanded the analysis by increasing the number of P. flavoviridis and P. tokarensis collected from 10 islands: Amami-Oshima (5 specimens), Kakeromajima (4) and Tokunoshima (4) from the Amami Islands, Okinawajima (4), Iheyajima (4), Iejima (4), Tokashikijima (4) and Kumejima (4) from the Okinawa Islands, Kodakarajima (P. tokarensis) (4) and Takarajima (P. tokarensis) (4) from the Tokara Islands. The maximum likelihood tree of the 44 samples replicated the significant divergence of P. flavoviridis between the Amami Clade including Amami-Oshima, Kakeromajima and Tokunoshima and the Okinawa Clade including Okinawajima, Iheyajima, Iejima, Tokashikijima and Kumejima. The Amami Clade also include all specimens from the Tokara Islands currently known as an independent species, P. tokarensis, suggesting the paraphyly of the taxon, P. flavoviridis. In contrast, we observed a distinct lineage of the two specimens from the Yaeyama Islands, supporting the validity of the taxon, P. elegans as an independent species. By MCMC method, we estimated the divergence time between the Amami Clade and the Okinawa Clade to be 6.51 MYA, suggesting that the vicariance of the two clades preceded the geological separation of the Amami Islands and the Okinawa Islands (~1.5 MYA). As expected from the limited mobility of terrestrial reptiles including snakes, we observed high genetic divergence in Habu mtDNA among Japanese subtropical island populations..
11. Ken Sano, Shiroh Miura, Toshiya Fujiwara, Ryuta Fujioka, Akiko Yorita, Kazuhito Noda, Hiroshi Kida, Koichi Azuma, Shinjiro Kaieda, Ken Yamamoto, Takayuki Taniwaki, Yasuyuki Fukumaki, Hiroki Shibata, A novel missense mutation of RYR1 in familial idiopathic hyper CK-emia, Journal of Neurological Sciences, 10.1016/j.jns.2015.06.035, 356, 1-2, 142-147, 2015.09, Persistent elevation of serum creatine kinase (CK) without any symptoms has been called idiopathic hyper CKemia
(IHCK).We examined a four-generation Japanese pedigree of familial IHCK. Themultipoint linkage analysis
of the pedigree showed seven clear peaks of logarithm of odds (LOD) scores (>1.4). By the exome sequencing
followed bymultiple filtering processes,we identified one novel heterozygous nonsynonymous single nucleotide
variant (SNV), c.7034GNC, p.S2345T in the ryanodine receptor 1 gene, RYR1 cosegregated with IHCK in the pedigree.
Mutation Taster predicted this substitution as “disease causing” (p = 0.999). The PolyPhen-2 and PANTHER
subPSEC scores for the substitution are 0.911 (possibly damaging) and−3.56 (probably damaging), respectively.
We confirmed the absence of the SNV in 511 healthy Japanese individuals excluding the possibility of a normal
variant with a very low frequency. Immunohistochemistry andWestern blotting of biopsy samples consistently
showed the expression level of RYR1 reduced in the patient. In real-time RT-PCR, the mRNA expression level of
RYR1 was also significantly reduced in the patient (p = 0.009). These results suggest that the novel
nonsynonymous SNV contribute to the vulnerability of the RYR1 protein through the dominant negative effect.
We conclude that the SNV in the RYR1 gene is one of the responsible genes of IHCK..
12. Naoki Kubo, Hidehiro Toh, Kenjiro Shirane, Takayuki Shirakawa, Hisato Kobayashi, Sato Tetsuya, Hidetoshi Sone, Yasuyuki Sato, Shin-ichi Tomizawa, Yoshinori Tsurusaki, Hiroki Shibata, Hirotomo Saitsu, Yutaka Suzuki, Naomichi Matsumoto, Tomohiro Kono, Kazuyuki Ohbo, Hiroyuki Sasaki, DNA methylation and gene expression dynamics during spermatogonial stem cell differentiation in the early postnatal mouse testis., BMC Genomics, 10.1186/s12864-015-1833-5, 16, 1, 624-624, 2015.08, Background: In the male germline, neonatal prospermatogonia give rise to spermatogonia, which include stem cell
population (undifferentiated spermatogonia) that supports continuous spermatogenesis in adults. Although the levels of
DNA methyltransferases change dynamically in the neonatal and early postnatal male germ cells, detailed genome-wide
DNA methylation profiles of these cells during the stem cell formation and differentiation have not been reported.
Results: To understand the regulation of spermatogonial stem cell formation and differentiation, we examined the DNA
methylation and gene expression dynamics of male mouse germ cells at the critical stages: neonatal prospermatogonia,
and early postntal (day 7) undifferentiated and differentiating spermatogonia. We found large partially methylated
domains similar to those found in cancer cells and placenta in all these germ cells, and high levels of non-CG
methylation and 5-hydroxymethylcytosines in neonatal prospermatogonia. Although the global CG methylation levels
were stable in early postnatal male germ cells, and despite the reported scarcity of differential methylation in the adult
spermatogonial stem cells, we identified many regions showing stage-specific differential methylation in and around
genes important for stem cell function and spermatogenesis. These regions contained binding sites for specific
transcription factors including the SOX family members.
Conclusions: Our findings show a distinctive and dynamic regulation of DNA methylation during spermatogonial stem
cell formation and differentiation in the neonatal and early postnatal testes. Furthermore, we revealed a unique
accumulation and distribution of non-CG methylation and 5hmC marks in neonatal prospermatogonia. These findings
contrast with the reported scarcity of differential methylation in adult spermatogonial stem cell differentiation and
represent a unique phase of male germ cell development..
13. Hiroki Shibata, Ken Yamamoto, Zhu Sun, Akira Oka, Hidetoshi Inoko, Tadao Arinami, Toshiya Inada, Hiroshi Ujike, Masanari Itokawa, Mamoru Tochigi, Yuichiro Watanabe, Toshiyuki Someya, Hiroshi Kunugi, Tatsuyo Suzuki, Nakao Iwata, Norio Ozaki, Yasuyuki Fukumaki, Genome-wide association study of schizophrenia using microsatellite markers in the Japanese population., Psychiatr Genetics, 23, 3, 117-123, 2013.06, OBJECTIVES: To search for schizophrenia susceptibility loci, we carried out a case-control study using 28601 microsatellite markers distributed across the entire genome.
MATERIALS AND METHODS: To control the highly multiple testing, we designed three sequential steps of screening using three independent sets of pooled samples, followed by the confirmatory step using an independent sample set (>2200 case-control pairs).
RESULTS: The first screening using pooled samples of 157 case-control pairs showed 2966 markers to be significantly associated with the disorder (PCONCLUSION: Genome-wide association study using microsatellite markers suggested SLC23A3, CNPPD1, and FAM134A genes as candidates for schizophrenia susceptibility in the Japanese population..
14. Hiroki Goto, Kazunori Watanabe, Naozumi Araragi, Rui Kageyama, Kunika Tanaka, Yoko Kuroki, Atsushi Toyoda, Masahira Hattori, Yoshiyuki Sakaki, Asao Fujiyama, Yasuyuki Fukumaki, Hiroki Shibata., The identification and functional implications of human-specific "fixed" amino acid substitutions in the glutamate receptor family., BMC Evolutionary Biology., 9: 224, 2009.09, [URL], Background: The glutamate receptors (GluRs) play a vital role in the mediation of excitatory synaptic transmission in the central nervous system. To clarify the evolutionary dynamics and mechanisms of the GluR genes in the lineage leading to humans, we determined the complete sequences of the coding regions and splice sites of 26 chimpanzee GluR genes.
Results: We found that all of the reading frames and splice sites of these genes reported in humans were completely conserved in chimpanzees, suggesting that there were no gross structural changes in humans after their divergence from the human-chimpanzee common ancestor. We observed low KA/KS ratios in both humans and chimpanzees, and we found no evidence of accelerated evolution. We identified 30 human-specific "fixed" amino acid substitutions in the GluR genes by analyzing 80 human samples of seven different populations worldwide. Grantham's distance analysis showed that GRIN2C and GRIN3A are the most and the second most diverged GluR genes between humans and chimpanzees. However, most of the substitutions are non-radical and are not clustered in any particular region. Protein motif analysis assigned 11 out of these 30 substitutions to functional regions. Two out of these 11 substitutions, D71G in GRIN3A and R727H in GRIN3B, caused differences in the functional assignments of these genes between humans and other apes.
Conclusion: We conclude that the GluR genes did not undergo drastic changes such as accelerated evolution in the human lineage after the divergence of chimpanzees. However, there remains a possibility that two human-specific "fixed" amino acid substitutions, D71G in GRIN3A and R727H in GRIN3B, are related to human-specific brain function.
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15. Hiroki Shibata, Ayako Tani, Tomoyuki Chikuhara, Rumiko Kikuta, Mayumi Sakai, Hideaki Ninomiya, Nobutada Tashiro, Nakao Iwata, Norio Ozaki and Yasuyuki Fukumaki. , Association study of polymorphisms in the group III metabotropic glutamate receptor genes, GRM4 and GRM7, with schizophrenia. , Psychiatric Res. , 167:88-96., 2009.04, Based on the hypothesis that a glutamatergic dysfunction is involved in the pathophysiology of schizophrenia, we have been conducting systematic studies on the association between glutamate receptor genes and schizophrenia. Here we report association studies of schizophrenia with polymorphisms in group III metabotropic glutamate receptor genes, GRM4 and GRM7. We selected 8 and 43 common SNPs distributed in the entire gene regions of GRM4 (> 111 kb) and GRM7 (> 900 kb), respectively. We scanned significant associations with schizophrenia using 100 case-control pairs of Japanese. We identified two neighboring SNPs (rs12491620 and rs1450099) in GRM7 showing highly significant haplotype association with schizophrenia surviving the FDR correction. We then performed additional typing of the two SNPs using the expanded sample set (404 cases and 420 controls) and confirmed the significant association with the disease. We conclude that at least one susceptibility locus for schizophrenia is located within or nearby GRM7, whereas GRM4 is unlikely to be a major susceptibility gene for schizophrenia in the Japanese population..
16. Shibata H, Aramaki T, Sakai M, Ninomiya H, Tashiro N, Iwata N, Ozaki N and Fukumaki Y., Association study of polymorphisms in the GluR7, KA1 and KA2 kainate receptor genes (GRIK3, GRIK4, GRIK5) with schizophrenia., Psychiatry Res., 141: 39-51, 2006.01.
17. Makino C, Shibata H, Ninomiya H, Tashiro N, Fukumaki Y., Identification of single-nucleotide polymorphisms in the human N-methyl-D-aspartate receptor subunit NR2D gene, GRIN2D, and association study with schizophrenia., Psychiatr Genet., 10.1097/00041444-200509000-00014, 15, 3, 215-221, 15 (3): 215-221., 2005.09.
18. Deng XD, Shibata H, Ninomiya H, Tashiro N, Iwata N, Ozaki N and Fukumaki Y., Association study of polymorphisms in the excitatory amino acid transporter 2 gene (SLC1A2) with schizophrenia., BMC Psychiatry, 10.1186/1471-244X-4-21, 4, 4(1): 21, 2004.08.
19. Takaki H, Kikuta R, Shibata H, Ninomiya H, Tashiro N and Fukumaki Y., Positive associations of polymorphisms in the metabotropic glutamate receptor type 8 gene (GRM8) with schizophrenia., Am J Med Genet., 10.1002/ajmg.b.20108, 128B, 1, 6-14, 128B: 6-14., 2004.07.
20. The Japanese Schizophrenia Sib-pair Linkage Group., Initial genome-wide scan for linkage with schizophrenia in the Japanese Schizophrenia Sib-pair Linkage Group (JSSLG) families., Am J Med Genet., 10.1002/ajmg.b.20022, 120B, 1, 22-28, 120B(1): 22-8., 2003.07.
21. Fujii Y, Shibata H, Kikuta R, Makino C, Tani A, Hirata N, Shibata A, Ninomiya H, Tashiro N, Fukumaki Y., Positive associations of polymorphisms in the metabotropic glutamate receptor type 3 gene (GRM3) with schizophrenia., Psychiatric Genetics., 10.1097/01.ypg.0000056682.82896.b0, 13, 2, 71-76, 13(2):71-6., 2003.06.
22. Makino C, Fujii Y, Kikuta R, Hirata N, Tani A, Shibata A, Ninomiya H, Tashiro N, Shibata H, Fukumaki Y., Positive association of the AMPA receptor subunit GluR4 gene (GRIA4) haplotype with schizophrenia: linkage disequilibrium mapping using SNPs evenly distributed across the gene region., Am J Med Genet., 10.1002/ajmg.b.10041, 116B, 1, 17-22, 116B (1): 17-22., 2003.01.
23. Shibata H, Shibata A, Ninomiya H, Tashiro N, Fukumaki Y., Association study of polymorphisms in the GluR6 kainate receptor gene (GRIK2) with schizophrenia., Psychiatry Res., 10.1016/S0165-1781(02)00231-7, 113, 1-2, 59-67, 113(1-2): 59-67., 2002.12.
24. Tani A, Kikuta R, Itoh K, Joo A, Shibata H, Ninomiya H, Tashiro N and Fukumaki Y., Polymorphism analysis of upstream regions of the human N-methyl-D-aspartate receptor subunit NR1 gene (GRIN1): implications for schizophrenia., Schizophr Research, 10.1016/S0920-9964(02)00161-5, 58, 1, 83-86, 58 (1):83-86., 2002.11.
25. Shibata H, Joo A, Fujii Y, Tani A, Makino C, Hirata N, Kikuta R, Ninomiya H, Tashiro N, Fukumaki Y., Association study of polymorphisms in the coding region of the GluR5 kainate receptor gene (GRIK1) with schizophrenia., Psychiatric Genetics, 10.1097/00041444-200109000-00005, 11, 3, 139-144, 11 (3): 139-144., 2001.09.
26. Kiehl TR, Shibata H, Huynh DP, Vo T, Pulst SM., Identification and expression of a mouse ortholog of A2BP1., Mammalian Genome., 12 (8): 595-601., 2001.08.
27. Shibata H, Huynh DP, Pulst SM., A novel protein with RNA binding motif binds to C-terminal ataxin-2., Human Molecular Genetics, 9 (9): 1303-1313., 2000.05.
28. Shibata H, Tahira T, Hayashi K., RNA-primed PCR., Genome Research, 10.1101/gr.5.4.400, 5, 4, 400-403, 5 (4): 400-403., 1995.11.
29. Shibata H, Yamazaki T., Molecular evolution of the duplicated Amy locus in the Drosophila melanogaster species subgroup: concerted evolution only in coding region and an excess of nonsynonymous substitutions in speciation., Genetics, 141, 1, 223-236, 141 (1): 223-236., 1995.09.
Presentations
1. Takuya Morikawa, Hiroaki Ohishi, Kengo Kosaka, Tomofumi Shimojo, Akihiro Nagano, Itsuki Taniguchi, Ryuta Fujioka, Kosei Moriyama, Motoko Unoki, Masatomo Takahashi, Motonao Nakao, Yoshihiro Izumi, Takeshi Bamba, Hiroyuki Sasaki, Shiroh Miura, Hiroki Shibata, Ddhd1 knockout mouse as a model for familial spastic paraplegia., 70th Annual Meeting of The American Society of Human Genetics, 2020.10.
2. Shiroh Miura, Tomofumi Shimojo, Ryuta Fujioka, Yusuke Uchiyama, Hiroki Shibata, A nonsense variant in the ARSD gene associated with young-onset Parkinson’s disease., The European Human Genetics Conference 2020, 2020.06.
3. Hiroki Shibata, Diversity and evolution of venom protein genes of a Japanese endemic pit viper, habu, Protobothrops flavoviridis., The 1st NUS-FU-KU Joint Symposium on Biochemistry in FUKUOKA, 2019.10.
4. Hiroki Shibata H, Naoko Oda-Ueda, Takahito Chijiwa, Hitomi Nakamura, Kazuaki Yamaguchi, Shosaku Hattori, Kazumi Matsubara, Yoichi Matsuda, Ryo Koyanagi, Yasuyuki Fukumaki, Motonori Ohno, Eiichi Shoguchi, Noriyuki Satoh, Tomohisa Ogawa, Habu snake (Protobothrops flavoviridis) genome study: Genomic architecture implicated in the multiplication and accelerated evolution of venom protein.v, EMBO/EMBL Symposium: Systems Genetics: From Genomes to Complex Traits, 2019.09.
5. Akiko Isomoto, Kanako Hisata, Jun Inoue, Eiichi Shoguchi, Noriyuki Satoh, Tomohisa Ogawa, Hiroki Shibata, Identification of non-venom protein genes highly expressed in the venom gland of habu (Protobothrops flavoviridis) and their potential contribution to the quality control of venom proteins., 20th World Congress of the International Society of Toxinology, 2019.09.
6. Hiroki Shibata, Whole genome sequencing of a Japanese endemic pit viper, habu, Protobothrops flavoviridis reveals accelerated evolution of venom protein genes enriched in microchromosomal regions., Integrative Organismal Biology Symposium: Integrating Structural Biology with ‘Omics in Asia-Pacific’, 2019.06.
7. Shiroh Miura, Kengo Kosaka, Ryuta Fujioka, Takayuki Taniwaki, Hiroki Shibata, Genetic analysis of autosomal dominant motor and sensory neuropathy with proximal dominancy in the lower extremities, urinary disturbance, and paroxysmal dry cough., The European Human Genetics Conference 2019, 2019.06.
8. Takuya Morikawa, Shiroh Miura, Hiroaki Ohishi, Kengo Kosaka, Tomofumi Shimojo, Akihiro Nagano, Ryuta Fujioka, Kosei Moriyama, Motoko Unoki, Masatomo Takahashi, Motonao Nakao, Yoshihiro Izumi, Takeshi Bamba, Hiroyuki Sasaki, Hiroki Shibata, Ddhd1 knockout mouse as a model for familial spastic paraplegia., The European Human Genetics Conference 2019, 2019.06.
9. Miura S, Shimojo T, Kosaka K, Fujioka R, Taniwaki T, Shibata H., Exome sequencing in sporadic patients with young-onset parkinsonism reveals novel candidates., 2018 American Neurological Association Annual Meeting, 2018.11.
10. Shibata H, Chijiwa T, Oda-Ueda N, Yamaguchi K, Hattori S, Matsubara K, Matsuda Y, Isomoto A, Koyanagi R, Hisata K, Fukumaki Y, Ohno M, Shoguchi E, Satoh N, Ogawa T. , Whole genome sequencing of a Japanese endemic pit viper, habu, Protobothrops flavoviridis reveals accelerated evolution of venom protein genes enriched in microchromosomal regions., The International Society of Toxinology 2018 European Section Congress, 2018.09.
11. Shibata H, Chijiwa T, Oda-Ueda N, Yamaguchi K, Hattori S, Matsubara K, Matsuda Y, Koyanagi R, Hisata K, Fukumaki Y, Ohno M, Shoguchi E, Satoh N, Ogawa T. , Whole genome sequencing of a Japanese endemic pit viper, habu, Protobothrops flavoviridis reveals accelerated evolution of venom protein genes enriched in microchromosomal regions., SMBE 2018, 2018.07.
12. Miura S, Kosaka K, Fujioka R, Uchiyama Y, Taniwaki T, Shimojo T, Shibata H., A novel nonsense variant (Lys177X) of FGF14 in a Japanese patient with autosomal dominant spinocerebellar ataxia., The European Human Genetics Conference 2018, 2018.06.
13. Shiroh Miura, Takuya Morikawa, Ryuta Fujioka, Kazuhito Noda, Kengo Kosaka, Takayuki Taniwaki, Hiroki Shibata, A novel missense variation (Q220R) of GNB4 encoding a guanine nucleotide-binding protein, beta-4 in a Japanese neuropathy family., The European Human Genetics Conference 2017, 2017.05.
14. Hiroki Shibata, Accelerated evolution of venom protein genes in the habu genome., Global Symposium GEM , 2017.03.
15. Tomohisa Ogawa, Takahito Chijiwa, Naoko Oda-Ueda, Hitomi Nakamura, Shousaku Hattori, Kazumi Matsubara, Yoichi Matsuda, Kazuki Mori, Kosuke Tashiro, Shinichi Yamasaki, Manabu Fujie, Hiroki Goto, Ryo Koyanagi, Yasuyuki Fukumaki, Motonori Ohn, Eiichi Shoguchi, Kanako Hisata, Noriyuki Satoh, Hiroki Shibata, Habu snake genome reveals the evolutionary strategy for generating the venom-related genes., Global Symposium GEM , 2017.03.
16. Takuya Morikawa, Shiroh Miura, Kengo Kosaka, Ryuta Fujioka, Ken Sano, Akiko Yorita, Kosuke Aoki, Yusuke Uchiyama, Takayuki Taniwaki, Hiroki Shibata, Heterozygous missense mutation in SEC24A encoding a coat protein complex II vesicle associated with autosomal dominant spinocerebellar ataxia., 66th Annual Meeting of The American Society of Human Genetics, 2016.10.
17. Ryuta Fujioka, Shiroh Miura, Kohei Yamada, Takuya Morikawa, Hiromichi Motooka, Yasuhiro Aso, Noriyuki Kimura, Ken Sano, Ken Yamamoto, Takayuki Taniwaki, Hiroki Shibata, A missense mutation in the PPIG gene encoding the peptidyl-prolyl isomerase G in patients with autosomal dominant benign adult familial myoclonic epilepsy., 66th Annual Meeting of The American Society of Human Genetics, 2016.10.
18. Whole genome sequencing of a Japanese endemic pit viper, habu, Protobothrops flavoviridis reveals venom-related genes enriched in microchromosomal regions..
19. 柴田 弘紀, 山本真由美, 千々岩崇仁, 服部正策, 上田直子, 大野素徳, 服巻 保幸, Genetic divergence of mitochondria genome sequence among island populations of Japanese Habu snakes, Protobothrops., 第35回日本分子生物学会年会, 2012.12.
20. 藤原敏弥, 柴田 弘紀, 三浦史郎, 山本真由美, 貴田浩志, 野田和人, 加來庸一郎, 岩城 明子, 綾部光芳, 谷脇考恭, 服巻 保幸, Linkage-Exome approach to identify the responsible variant for a novel type of hereditary neuropathy, 第35回日本分子生物学会年会, 2012.12.
21. Molecular Evolutionary Study Of The Ionotropic Glutamate-Receptor Gene Family As Schizophrenia Susceptibility Genes: Human-Specific Balancing Selection In GRIN2B Upstream Region.
Membership in Academic Society
  • The Herpetological Society of Japan
  • Society of Evolutionary Studies, Japan
  • The Genetics Society of Japan
  • The Molecular Biology Society of Japan
  • The American Society of Human Genetics
  • The Japan Society of Human Genetics