Updated on 2025/04/14

写真a

 
KUSAKABE,Rie
 
Organization
Faculty of Chemistry, Materials and Bioengineering Associate Professor
Title
Associate Professor
Profile

広島市生まれ。関西大学化学生命工学部准教授。京都大学理学部、カリフォルニア大学バークレー校にて学部時代を過ごし、北海道大学大学院理学研究科博士課程修了、博士(理学)。専門は動物の発生生物学・進化発生学。主な実験材料はメダカで、初期胚を使った実験、遺伝子工学的な手法を使って生命の謎に迫ろうとしている。

External link

Degree

  • 博士(理学)

Research Interests

  • 脊椎動物

  • メダカ

  • Da変異体

  • 軸上筋

  • 筋分化

  • 体節

  • 軸下筋

  • ヤツメウナギ

  • microRNA

  • 形態進化

  • 脊索動物

  • 機能性RNA

Research Areas

  • Life Science / Developmental biology

  • Life Science / Evolutionary biology

Education

  • Hokkaido University

    - 2000

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    Country: Japan

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Research History

  • Kansai University   Faculty of Chemistry , Materials and Bioengineering Department of Life Science and Biotechnology   Associate Professor

    2023.4

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  • Kansai University   Faculty of Chemistry , Materials and Bioengineering

    2020.4 - 2023.3

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  • 国立研究開発法人理化学研究所   形態進化研究チーム   研究員

    2014.5 - 2023.3

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  • Kobe University   Graduate School of Science

    2013.4 - 2014.4

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  • 神戸大学 理学(系)研究科(研究院)   助教

    2010.4

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  • Kobe University   Research Scientist

    2007.11 - 2010.3

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  • ?2003年3月 日本学術振興会特別研究員(PD)、2003年4月?独立行政法人理化学研究所研究員

    2000.4

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Professional Memberships

Papers

  • 円口類ヤツメウナギの遺伝子から探る脊椎動物の進化

    日下部りえ

    遺伝子医学   47   107 - 113   2024.1

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    Publishing type:Part of collection (book)  

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  • Thyroid and endostyle development in cyclostomes provides new insights into the evolutionary history of vertebrates Reviewed

    Wataru Takagi, Fumiaki Sugahara, Shinnosuke Higuchi, Rie Kusakabe, Juan Pascual-Anaya, Iori Sato, Yasuhiro Oisi, Nobuhiro Ogawa, Hiroshi Miyanishi, Noritaka Adachi, Susumu Hyodo, Shigeru Kuratani

    BMC Biology   20 ( 1 )   2022.12

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    Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media LLC  

    Abstract

    Background

    The endostyle is an epithelial exocrine gland found in non-vertebrate chordates (amphioxi and tunicates) and the larvae of modern lampreys. It is generally considered to be an evolutionary precursor of the thyroid gland of vertebrates. Transformation of the endostyle into the thyroid gland during the metamorphosis of lampreys is thus deemed to be a recapitulation of a past event in vertebrate evolution. In 1906, Stockard reported that the thyroid gland in hagfish, the sister cyclostome group of lampreys, develops through an endostyle-like primordium, strongly supporting the plesiomorphy of the lamprey endostyle. However, the findings in hagfish thyroid development were solely based on this single study, and these have not been confirmed by modern molecular, genetic, and morphological data pertaining to hagfish thyroid development over the last century.

    Results

    Here, we showed that the thyroid gland of hagfish undergoes direct development from the ventrorostral pharyngeal endoderm, where the previously described endostyle-like primordium was not found. The developmental pattern of the hagfish thyroid, including histological features and regulatory gene expression profiles, closely resembles that found in modern jawed vertebrates (gnathostomes). Meanwhile, as opposed to gnathostomes but similar to non-vertebrate chordates, lamprey and hagfish share a broad expression domain of Nkx2-1/2-4, a key regulatory gene, in the pharyngeal epithelium during early developmental stages.

    Conclusions

    Based on the direct development of the thyroid gland both in hagfish and gnathostomes, and the shared expression profile of thyroid-related transcription factors in the cyclostomes, we challenge the plesiomorphic status of the lamprey endostyle and propose an alternative hypothesis where the lamprey endostyle could be obtained secondarily in crown lampreys.

    DOI: 10.1186/s12915-022-01282-7

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    Other Link: https://link.springer.com/article/10.1186/s12915-022-01282-7/fulltext.html

  • Developmental Evolution of Hypaxial Muscles: Insights From Cyclostomes and Chondrichthyans Reviewed

    Rie Kusakabe, Masako Tanaka, Shigeru Kuratani

    Frontiers in Cell and Developmental Biology   9   2021.9

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    Jawed vertebrates possess two distinct groups of muscles in the trunk (epaxial and hypaxial muscles) primarily defined by the pattern of motor innervation from the spinal cord. Of these, the hypaxial group includes muscles with highly differentiated morphology and function, such as the muscles associated with paired limbs, shoulder girdles and tongue/infrahyoid (hypobranchial) muscles. Here we summarize the latest findings on the evolutionary mechanisms underlying the morphological variety of hypaxial musculature, with special reference to the molecular insights obtained from several living species that diverged early in vertebrate evolution. Lampreys, extant jawless vertebrates, lack many of derived traits characteristic of the gnathostomes, such as jaws, paired fins and epaxial/hypaxial distinction of the trunk skeletal musculatures. However, these animals possess the primitive form of the hypobranchial muscle. Of the gnathostomes, the elasmobranchs exhibit developmental mode of hypaxial muscles that is not identical to that of other gnathostomes in that the muscle primordia relocate as coherent cell aggregates. Comparison of expression of developmental genes, including Lbx genes, has delineated the temporal order of differentiation of various skeletal muscles, such as the hypobranchial, posterior pharyngeal and cucullaris (trapezius) muscles. We have proposed that the sequential addition of distal muscles, associated with expression of duplicated Lbx genes, promoted the elaboration of skeletal musculature. These analyses have revealed the framework of an evolutionary pathway that gave rise to the morphological complexity and diversity of vertebrate body patterns.

    DOI: 10.3389/fcell.2021.760366

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  • Developmental fates of shark head cavities reveal mesodermal contributions to tendon progenitor cells in extraocular muscles Reviewed

    Shunya Kuroda, Noritaka Adachi, Rie Kusakabe, Shigeru Kuratani

    Zoological Letters   2021.2

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    <jats:title>Abstract</jats:title><jats:p>Vertebrate extraocular muscles (EOMs) function in eye movements. The EOMs of modern jawed vertebrates consist primarily of four recti and two oblique muscles innervated by three cranial nerves. The developmental mechanisms underlying the establishment of this complex and the evolutionarily conserved pattern of EOMs are unknown. Chondrichthyan early embryos develop three pairs of overt epithelial coeloms called head cavities (HCs) in the head mesoderm, and each HC is believed to differentiate into a discrete subset of EOMs. However, no direct evidence of these cell fates has been provided due to the technical difficulty of lineage tracing experiments in chondrichthyans. Here, we set up an <jats:italic>in ovo</jats:italic> manipulation system for embryos of the cloudy catshark <jats:italic>Scyliorhinus torazame</jats:italic> and labeled the epithelial cells of each HC with lipophilic fluorescent dyes. This experimental system allowed us to trace the cell lineage of EOMs with the highest degree of detail and reproducibility to date. We confirmed that the HCs are indeed primordia of EOMs but showed that the morphological pattern of shark EOMs is not solely dependent on the early pattern of the head mesoderm, which transiently appears as tripartite HCs along the simple anteroposterior axis. Moreover, we found that one of the HCs gives rise to tendon progenitor cells of the EOMs, which is an exceptional condition in our previous understanding of head muscles; the tendons associated with head muscles have generally been supposed to be derived from cranial neural crest (CNC) cells, another source of vertebrate head mesenchyme. Based on interspecies comparisons, the developmental environment is suggested to be significantly different between the two ends of the rectus muscles, and this difference is suggested to be evolutionarily conserved in jawed vertebrates. We propose that the mesenchymal interface (head mesoderm vs CNC) in the environment of developing EOM is required to determine the processes of the proximodistal axis of rectus components of EOMs.</jats:p>

    DOI: 10.1186/s40851-021-00170-2

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  • Novel developmental bases for the evolution of hypobranchial muscles in vertebrates. Reviewed International journal

    Rie Kusakabe, Shinnosuke Higuchi, Masako Tanaka, Mitsutaka Kadota, Osamu Nishimura, Shigeru Kuratani

    BMC biology   18 ( 1 )   120 - 120   2020.9

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    BACKGROUND: Vertebrates are characterized by possession of hypobranchial muscles (HBMs). Cyclostomes, or modern jawless vertebrates, possess a rudimentary and superficial HBM lateral to the pharynx, whereas the HBM in jawed vertebrates is internalized and anteroposteriorly specified. Precursor cells of the HBM, marked by expression of Lbx1, originate from somites and undergo extensive migration before becoming innervated by the hypoglossal nerve. How the complex form of HBM arose in evolution is relevant to the establishment of the vertebrate body plan, but despite having long been assumed to be similar to that of limb muscles, modification of developmental mechanisms of HBM remains enigmatic. RESULTS: Here we characterize the expression of Lbx genes in lamprey and hagfish (cyclostomes) and catshark (gnathostome; jawed vertebrates). We show that the expression patterns of the single cyclostome Lbx homologue, Lbx-A, do not resemble the somitic expression of mammalian Lbx1. Disruption of Lbx-A revealed that LjLbx-A is required for the formation of both HBM and body wall muscles, likely due to the insufficient extension of precursor cells rather than to hindered muscle differentiation. Both homologues of Lbx in the catshark were expressed in the somitic muscle primordia, unlike in amniotes. During catshark embryogenesis, Lbx2 is expressed in the caudal HBM as well as in the abdominal rectus muscle, similar to lamprey Lbx-A, whereas Lbx1 marks the rostral HBM and pectoral fin muscle. CONCLUSIONS: We conclude that the vertebrate HBM primarily emerged as a specialized somatic muscle to cover the pharynx, and the anterior internalized HBM of the gnathostomes is likely a novelty added rostral to the cyclostome-like HBM, for which duplication and functionalization of Lbx genes would have been a prerequisite.

    DOI: 10.1186/s12915-020-00851-y

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  • Developmental mechanisms of migratory muscle precursors in medaka pectoral fin formation. Reviewed

    Tani-Matsuhana S, Kusakabe R, Inoue K

    Development genes and evolution   2018.7

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    DOI: 10.1007/s00427-018-0616-9

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  • The neural crest and evolution of the head/trunk interface in vertebrates Reviewed

    Shigeru Kuratani, Rie Kusakabe, Tatsuya Hirasawa

    Developmental Biology   2018

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Elsevier Inc.  

    The migration and distribution patterns of neural crest (NC) cells reflect the distinct embryonic environments of the head and trunk: cephalic NC cells migrate predominantly along the dorsolateral pathway to populate the craniofacial and pharyngeal regions, whereas trunk crest cells migrate along the ventrolateral pathways to form the dorsal root ganglia. These two patterns thus reflect the branchiomeric and somitomeric architecture, respectively, of the vertebrate body plan. The so-called vagal NC occupies a postotic, intermediate level between the head and trunk NC. This level of NC gives rise to both trunk- and cephalic-type (circumpharyngeal) NC cells. The anatomical pattern of the amphioxus, a basal chordate, suggests that somites and pharyngeal gills coexist along an extensive length of the body axis, indicating that the embryonic environment is similar to that of vertebrate vagal NC cells and may have been ancestral for vertebrates. The amniote-like condition in which the cephalic and trunk domains are distinctly separated would have been brought about, in part, by anteroposterior reduction of the pharyngeal domain.

    DOI: 10.1016/j.ydbio.2018.01.017

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  • Migratory appendicular muscles precursor cells in the common ancestor to all vertebrates. Reviewed International journal

    Eri Okamoto, Rie Kusakabe, Shigehiro Kuraku, Susumu Hyodo, Alexandre Robert-Moreno, Koh Onimaru, James Sharpe, Shigeru Kuratani, Mikiko Tanaka

    Nature ecology & evolution   1 ( 11 )   1731 - 1736   2017.11

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    In amniote embryos, skeletal muscles in the trunk are derived from epithelial dermomyotomes, the ventral margin of which extends ventrally to form body wall muscles. At limb levels, ventral dermomyotomes also generate limb-muscle precursors, an Lbx1-positive cell population that originates from the dermomyotome and migrates distally into the limb bud. In elasmobranchs, however, muscles in the paired fins were believed to be formed by direct somitic extension, a developmental pattern used by the amniote body wall muscles. Here we re-examined the development of pectoral fin muscles in catsharks, Scyliorhinus, and found that chondrichthyan fin muscles are indeed formed from Lbx-positive muscle precursors. Furthermore, these precursors originate from the ventral edge of the dermomyotome, the rest of which extends towards the ventral midline to form body wall muscles. Therefore, the Lbx1-positive, de-epithelialized appendicular muscle precursors appear to have been established in the body plan before the divergence of Chondrichthyes and Osteichthyes.

    DOI: 10.1038/s41559-017-0330-4

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  • Publisher Correction: Migratory appendicular muscles precursor cells in the common ancestor to all vertebrates. Reviewed International journal

    Eri Okamoto, Rie Kusakabe, Shigehiro Kuraku, Susumu Hyodo, Alexandre Robert-Moreno, Koh Onimaru, James Sharpe, Shigeru Kuratani, Mikiko Tanaka

    Nature ecology & evolution   1 ( 11 )   1784 - 1784   2017.11

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    In Fig. 2 of this Article originally published, some erroneous lines appeared on the left side of the images in panels c, e and g. The figure should have appeared as shown below. These errors have now been corrected in all versions of the Article.

    DOI: 10.1038/s41559-017-0374-5

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  • Developmental regulation and evolution of muscle-specific microRNAs Reviewed

    Rie Kusakabe, Kunio Inoue

    SEMINARS IN CELL & DEVELOPMENTAL BIOLOGY   47-48   9 - 16   2015.12

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    MicroRNAs (miRs) are a group of small RNAs that play a major role in post-transcriptional regulation of gene expression. In animals, many of the miRs are expressed in a conserved spatiotemporal manner. Muscle tissues, the major cellular systems involved in the locomotion and physiological functions of animals, have been one of the main sites for verification of miR targets and analysis of their developmental functions. During the determination and differentiation of muscle cells, numerous miRs bind to and repress target mRNAs in a highly specific but redundant manner. Interspecific comparisons of the sequences and expression of miRs have suggested that miR regulation became increasingly important during the course of vertebrate evolution. However, the detailed molecular interactions that have led to the highly complex morphological structures still await investigation. In this review, we will summarize the recent findings on the functional and developmental characteristics of miRs that have played major roles in vertebrate myogenesis, and discuss how the evolution of miRs is related to the morphological complexity of the vertebrates. (C) 2015 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.semcdb.2015.10.020

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  • Complete genome sequence of the mitochondrial DNA of the river lamprey, Lethenteron japonicum Reviewed

    Yuri L. Kawai, Kei Yura, Miyuki Shindo, Rie Kusakabe, Keiko Hayashi, Kenichiro Hata, Kazuhiko Nakabayashi, Kohji Okamura

    MITOCHONDRIAL DNA   26 ( 6 )   863 - 864   2015

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    Lampreys are eel-like jawless fishes evolutionarily positioned between invertebrates and vertebrates, and have been used as model organisms to explore vertebrate evolution. In this study we determined the complete genome sequence of the mitochondrial DNA of the Japanese river lamprey, Lethenteron japonicum, using next-generation sequencers. The sequence was 16,272 bp in length. The gene content and order were identical to those of the sea lamprey, Petromyzon marinus, which has been the reference among lamprey species. However, the sequence similarity was less than 90%, suggesting the need for the whole-genome sequencing of L. japonicum.

    DOI: 10.3109/19401736.2013.861432

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  • MiR-124 is Involved in Post-transcriptional Regulation of Polypyrimidine Tract Binding Protein 1 (PTBP1) During Neural Development in the Medaka, Oryzias latipes Reviewed

    Yumiko Kato, Rie Kusakabe, Kunio Inoue, Shin Tochinai

    ZOOLOGICAL SCIENCE   30 ( 11 )   891 - 900   2013.11

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    MicroRNAs (miRNAs) comprise a group of small noncoding RNA molecules thought to have contributed to the evolution of vertebrate brain homogeneity and diversity. The miRNA miR-124 is well conserved between invertebrates and vertebrates and is expressed abundantly in the central nervous system (CNS). We identified miR-124 in the medaka, Oryzias latipes, and investigated its role in neural development. The five candidate genes for medaka precursor miR-124 are unlinked on four different chromosomes and differ in nucleotide length. Their sequences suggest that they can generate functional miRNAs through conventional miRNA biogenesis by folding into stem-loop structures. Whole-mount in situ hybridization and northern blotting revealed that mature miR-124 is specifically expressed in the CNS and the eyes starting at two days post-fertilization. We also examined the sequences and expression of medaka Polypyrimidine tract binding protein 1 (Ptbp1), a possible direct target of miR-124. The 3'UTR of medaka Ptbp1 contains predicted binding motifs (target sites) for miR-124. A GFP reporter assay for the target sites or the entire 3'UTR showed that exogenous miR-124 silences PTBP1 expression in vivo. Our study suggests that medaka miR-124 is involved in post-transcriptional regulation of target genes in neural development and that medaka miR-124 homologs may have spatiotemporal roles different from those in other vertebrates.

    DOI: 10.2108/zsj.30.891

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  • Body wall development in lamprey and a new perspective on the origin of vertebrate paired fins Reviewed

    Frank J. Tulenko, David W. McCauley, Ethan L. MacKenzie, Sylvie Mazan, Shigeru Kuratani, Fumiaki Sugahara, Rie Kusakabe, Ann C. Burke

    Proceedings of the National Academy of Sciences of the United States of America   110 ( 29 )   11899 - 11904   2013.7

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    Classical hypotheses regarding the evolutionary origin of paired appendages propose transformation of precursor structures (gill arches and lateral fin folds) into paired fins. During development, gnathostome paired appendages form as outgrowths of body wall somatopleure, a tissue composed of somatic lateral plate mesoderm (LPM) and overlying ectoderm. In amniotes, LPM contributes connective tissue to abaxial musculature and forms ventrolateral dermis of the interlimb body wall. The phylogenetic distribution of this character is uncertain because lineage analyses of LPM have not been generated in anamniotes. We focus on the evolutionary history of the somatopleure to gain insight into the tissue context in which paired fins first appeared. Lampreys diverged from other vertebrates before the acquisition of paired fins and provide a model for investigating the preappendicular condition. We present vital dye fate maps that suggest the somatopleure is eliminated in lamprey as the LPM is separated from the ectoderm and sequestered to the coelomic linings during myotome extension. We also examine the distribution of postcranial mesoderm in catshark and axolotl. In contrast to lamprey, our findings support an LPM contribution to the trunk body wall of these taxa, which is similar to published data for amniotes. Collectively, these data lead us to hypothesize that a persistent somatopleure in the lateral body wall is a gnathostome synapomorphy, and the redistribution of LPM was a key step in generating the novel developmental module that ultimately produced paired fins. These embryological criteria can refocus arguments on paired fin origins and generate hypotheses testable by comparative studies on the source, sequence, and extent of genetic redeployment.

    DOI: 10.1073/pnas.1304210110

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  • Characterization of the compact bicistronic microRNA precursor, miR-1/miR-133, expressed specifically in Ciona muscle tissues Reviewed

    Rie Kusakabe, Saori Tani, Koki Nishitsuji, Miyuki Shindo, Kohji Okamura, Yuki Miyamoto, Kenta Nakai, Yutaka Suzuki, Takehiro G. Kusakabe, Kunio Inoue

    GENE EXPRESSION PATTERNS   13 ( 1-2 )   43 - 50   2013.1

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    Muscle-specific miR-1/206 and miR-133 families have been suggested to play fundamental roles in skeletal and cardiac myogenesis in vertebrates. To gain insights into the relationships between the divergence of these miRs and muscular tissue types, we investigated the expression patterns of miR-1 and miR-133 in two ascidian Ciona species and compared their genomic structures with those of other chordates. We found that Ciona intestinalis and Ciona savignyi each possess a single copy of the miR-1/miR-133 cluster, which is only 350 nucleotide long. During embryogenesis, Ciona miR-1 and miR-133 are generated as a single continuous primary transcript accumulated in the nuclei of the tail muscle cells, starting at the gastrula stage. In adults, mature miR-133 and miR-1 are differentially expressed in the heart and body wall muscle. Expression of the reporter gene linked to the 850-bp upstream region of the predicted transcription start site confirmed that this region drives the muscle-specific expression of the primary transcript of miR-1/miR-133. In many deuterostome lineages, including that of Ciona, the miR-1/133 cluster is located in the same intron of the mind bomb (mib) gene in reverse orientation. Our results suggest that the origin of genomic organization and muscle-specific regulation of miR-1/133 can be traced back to the ancestor of chordates. Duplication of this miR cluster might have led to the remarkable elaboration in the morphology and function of skeletal muscles in the vertebrate lineage. (C) 2012 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.gep.2012.11.001

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  • Developmental expression and evolution of muscle-specific microRNAs conserved in vertebrates Reviewed

    Tani, Saori, Kuraku, Shigehiro, Sakamoto, Hiroshi, Inoue, Kunio, Kusakabe, Rie

    Evolution & Development   15 ( 4 )   293 - 304   2013

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    DOI: 10.1111/ede.12039

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  • Expression and interaction of muscle-related genes in the lamprey imply the evolutionary scenario for vertebrate skeletal muscle, in association with the acquisition of the neck and fins. Reviewed International journal

    Rie Kusakabe, Shigehiro Kuraku, Shigeru Kuratani

    Developmental biology   350 ( 1 )   217 - 27   2011.2

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    Gnathostomes (jawed vertebrates) possess skeletal muscles with unique functional and developmental features that are absent from cyclostomes-i.e., lamprey and hagfish. These gnathostome-specific traits include the epaxial and hypaxial division of myotomes, paired fin/limb muscles, shoulder girdle muscles, and the muscle associated with the tongue and the neck. Many of these muscles are derived from several rostral somites, specifically from their hypaxial myotomic domains. However, it has not been clarified how the complicated morphology of these muscles was acquired in the evolution of vertebrates. Here we describe the expression of lamprey homologs of transcription factor genes, including a myogenic regulatory factor of the Myod family (MRF), Pax3/7, Lbx, and Zic, which play important roles in the development of ep-/hypaxial somitic muscles in gnathostomes, and show that the ventral portion of lamprey somites is comparable to the ventral dermomyotome in gnathostomes. The supra- and infraoptic muscles, derived from the two anterior somites in the lamprey, are molecularly specified before their extensive invasion into the head region. Of these, the infraoptic myotomes are suggested to represent the cucullaris homologue in the lamprey based on their topographical position in the embryonic pattern. Slightly caudal myotomes in the lamprey give rise to the hypobranchial muscle, the developmental homologue of the gnathostome hypobranchial musculature. The dorsal moieties of the lamprey somites express a Zic gene, which in teleosts specifies the epaxial identities of the somites. These evidences suggest that, although the myotomes in the ancestral jawless vertebrates do not exhibit ep-/hypaxial distinction at the morphological level, their dorsoventral specification would have already been present at gene regulatory levels, prior to the cyclostome-gnathostome divergence, which may have functioned as the key innovation to establish the ep-/hypaxial distinction in gnathostomes.

    DOI: 10.1016/j.ydbio.2010.10.029

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  • Genomic organization and embryonic expression of miR-430 in medaka (Oryzias latipes): Insights into the post-transcriptional gene regulation in early development (vol 449, pg 41, 2010) Reviewed

    Saori Tani, Rie Kusakabe, Kiyoshi Naruse, Hiroshi Sakamoto, Kunio Inoue

    GENE   457 ( 1-2 )   50 - 51   2010.6

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    DOI: 10.1016/j.gene.2010.02.010

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  • Genomic organization and embryonic expression of miR-430 in medaka (Oryzias latipes): Insights into the post-transcriptional gene regulation in early development Reviewed

    Saori Tani, Rie Kusakabe, Kiyoshi Naruse, Hiroshi Sakamoto, Kunio Inoue

    GENE   449 ( 1-2 )   41 - 49   2010.1

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    MicroRNAs (miRNAs, miRs) are short noncoding RNA molecules that negatively control the target mRNAs by binding to the 3&apos; untranslated region (UTR). Previous studies have demonstrated that miR-430 is encoded by a clustered multigene family and is abundantly expressed in early development. In zebrafish, miR-430 is needed to suppress primordial germ cell (PGC)-specific genes, such as nanos1, in somatic cells. However, the molecular characteristics of the miR-430 family in other teleost species have not been reported, and it is unclear whether such a function of miR-430 in PGC specification is a conserved feature of animals or not. In medaka (Oryzias latipes), a distantly related teleost, it has been suggested that PGC might be established in a different mode of specification from that of zebrafish. We characterized 16 miR-430 precursors in the medaka genomic sequence. These miR-430 genes form clusters on chromosome 4, which might share its evolutionary origin with that of the very large miR-430 clusters in zebrafish chromosome 4. However, none of the medaka miR-430 genes are identical to the zebrafish miR-430 paralogs. Medaka miR-430 expression starts during epiboly and decreases after axis formation. Functional analysis using reporter gene constructs showed that miR-430 repressed protein expression by binding to the 3&apos;UTR of zebrafish TDRD7. Consistently, the 3&apos;UTR of medaka TDRD7 contains at least two significant candidates for the putative miR-430 binding site. The ubiquitous and early expression of medaka miR-430 and its ability to downregulate GFP:TDRD7 reporter mRNA imply that miR-430 has a conserved role in early embryogenesis. Smaller copy numbers of miR-430 genes and relatively brief expression in medaka might represent the characteristics of this miRNA family in the common ancestor of teleosts. Changes in the relationships between miR-430 and the target mRNA might be related to differences in the localization patterns of PGC-related genes in medaka and zebrafish. (C) 2009 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.gene.2009.09.005

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  • Evolution of vertebrate skeletal myogenesis: Insights from the cyclostome lamprey Reviewed

    Kusakabe, Rie, Kuraku, Shigehiro, Kuratani, Shigeru

    Developmental Biology   344 ( 1 )   2010

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    DOI: 10.1016/j.ydbio.2010.05.401

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  • Evolution of vertebrate myogenesis, with special reference to the morphological variety of skeletal muscles Reviewed

    Kusakabe, Rie, Kuraku, Shigehiro, Kuratani, Shigeru, Inoue, Kunio

    Mechanisms of Development   126   2009

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    DOI: 10.1016/j.mod.2009.06.651

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  • Expression of Sox and Fibrillar Collagen Genes in Lamprey Larval Chondrogenesis With Implications for the Evolution of Vertebrate Cartilage Reviewed

    Kaoru Ohtanii, Tuoya Yao, Mari Kobayashi, Rie Kusakabe, Shigeru Kuratani, Hiroshi Wada

    JOURNAL OF EXPERIMENTAL ZOOLOGY PART B-MOLECULAR AND DEVELOPMENTAL EVOLUTION   310B ( 7 )   596 - 607   2008.11

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    Lampreys possess unique types of cartilage in which elastin-like proteins are the dominant matrix component, whereas gnathostome cartilage is mainly composed of fibrillar collagen. Despite the differences in protein composition, the Sox-col2a1 genetic cascade was suggested to be conserved between lamprey pharyngeal cartilage and gnathostome cartilage. We examined whether the cascade is conserved in another type of lamprey cartilage, the trabecular cartilage. We found that SoxD and SoxE are expressed in both trabecular and pharyngeal cartilages. However, trabecular cartilage shows no clade A fibrillar collagen gene expression, including genes expressed in pharyngeal cartilage of this animal. On the basis of these observations, we propose that lampreys possess an ancestral type of cartilage that is similar to amphioxus gill cartilage, and in this respect, gnathostome cartilage can be regarded as derived for the loss of elastin-like protein as a cartilage component and recruitment of fibrillar collagen, which is included as a minor component in the ancestral cartilage, as the main component. J. Exp. Zool. (Mol. Dev. Evol.) 310B:596-607, 2008. (C) 2008 Wiley-Liss, Inc.

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  • Identification of four Engrailed genes in the Japanese lamprey, Lethenteron japonicum. Reviewed International journal

    Manami Matsuura, Hidenori Nishihara, Koh Onimaru, Nobuhiro Kokubo, Shigehiro Kuraku, Rie Kusakabe, Norihiro Okada, Shigeru Kuratani, Mikiko Tanaka

    Developmental dynamics : an official publication of the American Association of Anatomists   237 ( 6 )   1581 - 9   2008.6

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    We have isolated four homologs of Engrailed genes from the Japanese lamprey, Lethenteron japonicum, an agnathan that occupies a critical phylogenic position between cephalochordates and gnathostomes. We named these four genes LjEngrailedA, LjEngrailedB, LjEngrailedC, and LjEngrailedD. LjEngrailedA, LjEngrailedB, and LjEngrailedD share a major expression domain in the presumptive midbrain-hindbrain boundary region of the central nervous system, although their levels and timing of expression differed. On the other hand, LjEngrailedC transcripts were in the pharyngeal ectoderm and the ventral ectoderm of the body wall. In addition, LjEngrailedA was expressed in the ventral side of the epibranchial muscle precursors. LjEngrailedD transcripts were seen in the mesodermal cells of the mandibular arch and later in a group of cells responsible for the formation of the upper lip, lower lip, and velum. Our results provide clues to the evolution of these structures as well as a possible scenario for duplication events of Engrailed genes.

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  • Identification and developmental expression of two Tbx1/10-related genes in the agnathan Lethenteron japonicum. Reviewed International journal

    Eva Tiecke, Manami Matsuura, Nobuhiro Kokubo, Shigehiro Kuraku, Rie Kusakabe, Shigeru Kuratani, Mikiko Tanaka

    Development genes and evolution   217 ( 10 )   691 - 7   2007.10

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    We have identified two Tbx1/10-related genes, LjTbx1/10A and LjTbx1/10B, from the Japanese river lamprey Lethenteron japonicum. We used in situ hybridization to characterize their expression pattern during embryonic development. LjTbx1/10A and LjTbx1/10B shared common expression in the pharyngeal arches and otic vesicle, although their levels and timing of expression differed markedly. LjTbx1/10A was highly expressed in the mesodermal core of pharyngeal arches and the adjacent endoderm throughout pharyngeal arch development, whereas LjTbx1/10B expression was only transiently upregulated in forming pharyngeal pouches. LjTbx1/10A transcripts first appeared at stage 25 in otic vesicles, whereas LjTbx1/10B transcripts could already be detected in the developing otic placode at stage 20. These results suggest that lamprey LjTbx1/10A and LjTbx1/10B may play distinct roles in the patterning and development of the pharyngeal apparatus. It appears that lamprey Tbx1/10 genes have undergone subfunctionalization independent from gnathostomes, with regard to both regulation and function.

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  • Evolutionary perspectives from development of mesodermal components in the lamprey Reviewed

    Rie Kusakabe, Shigeru Kuratani

    DEVELOPMENTAL DYNAMICS   236 ( 9 )   2410 - 2420   2007.9

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    Lampreys, a jawless vertebrate species, lack not only jaws but also several other organs, including ventral migratory muscles shared by gnathostomes. In the lamprey embryo, the mesoderm consists primarily of unsegmented head mesoderm, segmented somites, and yet uncharacterized lateral plate mesoderm, as in gnathostomes. Although the adult lamprey possesses segmented myotomes in the head, the head mesoderm of this animal is primarily unsegmented, similar to that in gnathostomes. In the trunk, the large part of lamprey somites is destined to form myotomes, and the Pax317 gene expression domain in the lateral part of somites is suggested to represent a dermomyotome homologue. Lamprey myotomes are not segregated by a horizontal myoseptum, which has been regarded as consistent with the apparent absence of a migratory population of hypaxial muscles shared by gnathostomes. However, recent analysis suggests that lampreys have established the gene regulatory cascade necessary for the ventrally migrating myoblasts, which functions in part during the development of the primordial hypobranchial muscle. There have also been new insights on the developmental cascade of lamprey cartilages, in which the Sox family of transcription factors plays major roles, as in gnathostomes. Thus, mesoderm development in lampreys may represent the ancestral state of gene regulatory mechanisms required for the evolution of the complex and diverse body plan of gnathostomes.

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  • Hox gene expression patterns in Lethenteron japonicum embryos--insights into the evolution of the vertebrate Hox code. Reviewed International journal

    Yoko Takio, Shigehiro Kuraku, Yasunori Murakami, Massimo Pasqualetti, Filippo M Rijli, Yuichi Narita, Shigeru Kuratani, Rie Kusakabe

    Developmental biology   308 ( 2 )   606 - 20   2007.8

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    The Hox code of jawed vertebrates is characterized by the colinear and rostrocaudally nested expression of Hox genes in pharyngeal arches, hindbrain, somites, and limb/fin buds. To gain insights into the evolutionary path leading to the gnathostome Hox code, we have systematically analyzed the expression pattern of the Hox gene complement in an agnathan species, Lethenteron japonicum (Lj). We have isolated 15 LjHox genes and assigned them to paralogue groups (PG) 1-11, based on their deduced amino acid sequences. LjHox expression during development displayed gnathostome-like spatial patterns with respect to the PG numbers. Specifically, lamprey PG1-3 showed homologous expression patterns in the rostral hindbrain and pharyngeal arches to their gnathostome counterparts. Moreover, PG9-11 genes were expressed specifically in the tailbud, implying its posteriorizing activity as those in gnathostomes. We conclude that these gnathostome-like colinear spatial patterns of LjHox gene expression can be regarded as one of the features already established in the common ancestor of living vertebrates. In contrast, we did not find evidence for temporal colinearity in the onset of LjHox expression. The genomic and developmental characteristics of Hox genes from different chordate species are also compared, focusing on evolution of the complex body plan of vertebrates.

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  • Insights into the sequential evolutionary events toward the acquisition of paired fins

    Manami Matsuura, Eva Tiecke, Rie Kusakabe, Shigeru Kuratani, Mikiko Tanaka

    ZOOLOGICAL SCIENCE   23 ( 12 )   1184 - 1185   2006.12

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  • Evolutionary embryology resurrected in Japan with a new molecular basis: Nori Satoh and the history of ascidian studies originating in Kyoto during the 20th century Reviewed

    S Kuratani, H Wada, R Kusakabe, K Agata

    INTERNATIONAL JOURNAL OF DEVELOPMENTAL BIOLOGY   50 ( 5 )   451 - 454   2006

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    This article briefly summarizes the scientific contributions of Nori Satoh, the winner of the 2005 edition of the Kowalevsky Medal, to Developmental Biology and especially to Evo-Devo with his 30 years of research on tunicates - a primitive chordate species. His research began with his pure developmental interest in the clock mechanism of cell differentiation and later expanded into various aspects of evolutionary and developmental phenomena. He is not only known as a founder of molecular biology-based tunicate studies, but also for his world-wide service to education and his prestigious publications in international scientific journals.

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  • Evolutionary embryology revived in Japan with a new molecular basis--Nori Satoh and history of ascidian studies in Kyoto in the 20th century Reviewed

    Kuratani, S., Kusakabe, R., Wada, H., Agata, K.

    Ontogenez   37 ( 6 )   2006

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  • Evolution and developmental patterning of the vertebrate skeletal muscles: Perspectives from the lamprey Reviewed

    R Kusakabe, S Kuratani

    DEVELOPMENTAL DYNAMICS   234 ( 4 )   824 - 834   2005.12

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    The myotome in gnathostome vertebrates, which gives rise to the trunk skeletal muscles, consists of epaxial (dorsal) and hypaxial (ventral) portions, separated by the horizontal myoseptum. The hypaxial portion contains some highly derived musculature that is functionally as well as morphologically well differentiated in all the gnathostome species. In contrast, the trunk muscles of agnathan lampreys lack these distinctions and any semblance of limb muscles. Therefore, the lamprey myotomes probably represent a primitive condition compared with gnathostomes. In this review, we compare the patterns of expression of some muscle-specific genes between the lamprey and gnathostomes. Although the cellular and tissue morphology of lamprey myotomes seems uniform and undifferentiated, some of the muscle-specific genes are expressed in a spatially restricted manner. The lamprey Pax3/7 gene, a cognate of gnathostome Pax3, is expressed only at the lateral edge of the myotomes and in the hypobranchial muscle, which we presume is homologous to the gnathostome hypobranchial muscle. Thus, the emergence of some part of a hypaxial-specific gene regulatory cascade might have evolved before the agnathan/gnathostome divergence, or before the evolutionary separation of epaxial and hypaxial muscles.

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  • Developmental fate of the mandibular mesoderm in the lamprey, Lethenteron japonicum: Comparative morphology and development of the gnathostome jaw with special reference to the nature of the trabecula cranii. Reviewed

    Kuratani S, Murakami Y, Nobusada Y, Kusakabe R, Hirano S

    Journal of experimental zoology. Part B, Molecular and developmental evolution   302 ( 5 )   458 - 468   2004.9

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  • Developmental fate of the mandibular mesoderm in the lamprey, Lethenteron japonicum: Comparative morphology and development of the gnathostome jaw with special reference to the nature of the trabecula cranii Reviewed

    S Kuratani, Y Murakami, Y Nobusada, R Kusakabe, S Hirano

    JOURNAL OF EXPERIMENTAL ZOOLOGY PART B-MOLECULAR AND DEVELOPMENTAL EVOLUTION   302B ( 5 )   458 - 468   2004.9

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    The vertebrate jaw is a mandibular-arch derivative, and is regarded as the synapomorphy that defines the gnathostomes. Previous studies (Kuratani et al., Phil. Trans. Roy. Soc. 356:15, 2001; Shigetani et al., Science 296:1319, 2002) have suggested that the oral apparatus of the lamprey is derived from both the mandibular and premandibular regions, and that the jaw has arisen as a secondary narrowing of the oral patterning mechanism into the mandibular-arch domain. The heterotopy theory of jaw evolution states that the lamprey upper lip is a premandibular element, leaving further questions unanswered as to the homology of the trabecula in the lamprey and gnathostomes, and to the morphological nature of the muscles in the upper lip. Using focal injection of vital dyes into the cheek process core of lamprey embryos, we found that the upper lip muscle and trabecula are both derived from mandibular mesoderm. Secondary movement of the muscle primordium is also evident when the expression of the early muscle marker gene, LjMA2, is visualized. A nerve-fiber labeling study revealed that the upper lip muscle-innervating neurons are located in the rostral part of the brain stem, where the trigeminal motor nuclei are not found in gnathostomes. We conclude that the lamprey upper lip is composed of premandibular ectomesenchyme and a lamprey-specific muscle component derived from the mandibular mesoderm innervated by lamprey-specific motoneurons. Furthermore, the lamprey trabecula is most likely equivalent to a mesodermally derived neurocranial element, similar to the parachordal element in gnathostomes, rather than to the neural-crest-derived prechordal element. (C) 2004 Wiley-Liss, Inc.

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  • Lamprey contractile protein genes mark different populations of skeletal muscles during development

    R Kusakabe, M Takechi, S Tochinai, S Kuratani

    JOURNAL OF EXPERIMENTAL ZOOLOGY PART B-MOLECULAR AND DEVELOPMENTAL EVOLUTION   302B ( 2 )   121 - 133   2004.3

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    Agnathan lampreys retain ancestral characteristics of vertebrates in the morphology of skeletal muscles derived from two mesodermal regions: trunk myotomes and unsegmented head mesoderm. During lamprey development, some populations of myoblasts migrate via pathways that differ from those of gnathostomes. To investigate the evolution of skeletal muscle differentiation in vertebrates, we characterize multiple contractile protein genes expressed in the muscle cells of the Japanese lamprey, Lethenteron japonicum. Lamprey actin gene LjMA2, and myosin heavy chain (MyHC) genes LjMyHC1 and LjMyHC2 are all expressed in the developing skeletal muscle cells of early embryos. However, LjMyHC1 and LjMyHC2 are expressed only in cells originating from myotomes, while LjMA2 is expressed in both myotomal and head musculature. Thus, in lampreys, myotomes and head mesoderm differ in the use of genes encoding contractile protein isoforms. Phylogenetic tree analyses including lamprey MyHCs suggest that the variety of muscle MyHC isoforms in different skeletal muscles may correspond to the morphological complexity of skeletal muscles of different vertebrate species. Another lamprey actin gene LjMA1 is likely to be the first smooth muscle actin gene isolated from non-tetrapods. We conclude that, in vertebrate evolution, the different regulatory systems for striated and smooth muscle-specific genes may have been established before the agnathan/gnathostome divergence. J. Exp. Zool. (Mol. Dev. Evol.) 302B:121-133, 2004. (C) 2004 Wiley-Liss, Inc.

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  • Lamprey contractile protein genes mark different populations of skeletal muscles during development. Reviewed

    Kusakabe R, Takechi M, Tochinai S, Kuratani S

    Journal of experimental zoology. Part B, Molecular and developmental evolution   302 ( 2 )   121 - 133   2004.3

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  • Origin of the vertebrate visual cycle: Genes encoding retinal photoisomerase and two putative visual cycle proteins are expressed in whole brain of a primitive chordate Reviewed

    Y Nakashima, T Kusakabe, R Kusakabe, A Terakita, Y Shichida, M Tsuda

    JOURNAL OF COMPARATIVE NEUROLOGY   460 ( 2 )   180 - 190   2003.5

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    The absorption of light by rhodopsin leads to the cis-to-trans isomerization of the chromophore to generate all-trans-retinal. In the visual cycle, the resultant all-trans-retinal is converted back into the 11-cis-retinal. In the mammalian eye, the retinal pigment epithelium (RPE) plays an essential role in the visual cycle. We have identified cDNA clones encoding three putative visual cycle proteins, homologs of mammalian retinal G-protein-coupled receptor (RGR), cellular retinaldehyde-binding protein (CRALBP) and beta-carotene 15,15'-monooxygenase (BCO)/ RPE65 in a primitive chordate, ascidian Ciona intestinalis. The mRNAs for these proteins are specifically expressed in the central nervous system during embryonic development. In the larva, the transcripts were widely distributed in the brain vesicle and visceral ganglion. Since visual pigment, Ci-opsin1, is solely expressed in photoreceptor cells, the visual cycle in this primitive chordate may take place in two compartments, which are coupled into a cycle by the direct flow of retinoids though the intercellular matrix. The Ci-opsin3, an ascidian homolog of mammalian RGR, was expressed in HEK 293S cells and purified after binding of retinal. The chromophore of Ci-opsin3 is in an all-trans-retinal and it is isomerized to an 11-cis-form upon absorption of light. Mammalian CRALBP and BCO/RPE65 are believed to play critical roles in the process of reisomerization of all-trans-retinoid to 11-cis-retinoid in RPE. The present data suggest that isomerization of all-trans-retinoid to 11-cis-retinoid occurs in the brain vesicle and visceral ganglion of a primitive chordate. J. Comp. Neurol. 460:180-190, 2003. (C) 2003 Wiley-Liss, Inc.

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  • Expression of foreign genes in lamprey embryos: an approach to study evolutionary changes in gene regulation. Reviewed

    Kusakabe R, Tochinai S, Kuratani S

    Journal of experimental zoology. Part B, Molecular and developmental evolution   296 ( 1 )   87 - 97   2003.4

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  • Expression of foreign genes in lamprey embryos: An approach to study evolutionary changes in gene regulation Reviewed

    R Kusakabe, S Tochinai, S Kuratani

    JOURNAL OF EXPERIMENTAL ZOOLOGY PART B-MOLECULAR AND DEVELOPMENTAL EVOLUTION   296B ( 1 )   87 - 97   2003.4

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    Evolution in development can be viewed as a sequence of changes in gene regulation. To investigate the cross-species compatibility of 5' upstream regulatory regions, we introduced exogenous gene constructs derived from a gnathostome genome into fertilized eggs of the Japanese lamprey, Lampetra japonica, a sister group of the gnathostomes. Eggs were injected with gene constructs in which a sequence encoding the green fluorescent protein (GFP) had been located downstream of either a virus promoter or 5' regulatory regions of medaka actin genes. Reporter gene expression was recorded for more than a month starting two days after injection. Although the expression patterns were highly mosaic and differed among individuals, GFP was expressed predominantly in the striated muscles of lamprey embryos when driven by the 5' upstream regions of the medaka muscle actin genes. This implies that a pan-vertebrate muscle-specific gene regulatory mechanism may have evolved before the agnathan/gnathostome divergence. This gene-transfer technique potentially facilitates the visualization of cells in various differentiating tissues throughout development. The introduction of developmental genes of the lamprey or other animals into lamprey embryos is another potentially important application, one that could provide us with information on the evolutionary changes in functions of genes or gene cascades. (C) 2003 Wiley-Liss, Inc.

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  • SYSTEMATIC ANALYSIS OF CIS-REGULATORY REGIONS OF CNS-SPECIFIC GENES IN THE ASCIDIAN LARVA : II. IN VIVO FUNCTIONAL ANALYSIS(Developmental Biology,Abstracts of papers presented at the 74^<th> Annual Meeting of the Zoological Society of Japan) :

    Yoshida Reiko, Niimi Takahiro, Sakurai Daisuke, Kusakabe Rie, Horie Takeo, Tsuda Motoyuki, Kusakabe Takehiro

    Zoological science   20 ( 12 )   1561 - 1561   2003

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  • Gene expression profiles in tadpole larvae of Ciona intestinalis Reviewed

    T Kusakabe, R Yoshida, Kawakami, I, R Kusakabe, Y Mochizuki, L Yamada, T Shin-i, Y Kohara, N Satoh, M Tsuda, Y Satou

    DEVELOPMENTAL BIOLOGY   242 ( 2 )   188 - 203   2002.2

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    A set of 12,779 expressed sequence tags (ESTs), both the 5'-most and T-most ends, derived from Ciona intestinalis tadpole larvae was categorized into 3521 independent clusters, from which 1013 clusters corresponding to 9424 clones were randomly selected to analyze genetic information and gene expression profiles. When compared with sequences in databases, 545 of the clusters showed significant matches (P &lt; E-15) with reported proteins, while 153 showed matches with putative proteins for which there is not enough information to categorize their function, and 315 had no significant sequence similarities to known proteins. Sequence-similarity analyses of the 545 clusters in relation to the biological functions demonstrated that 407 of them have functions that many kinds of cells use, 104 are associated with cell-cell communication, and 34 are transcription factors or other gene-regulatory proteins. Sequence prevalence distribution analysis demonstrated that more than one-half of the mRNAs are rate mRNAs. All of the 1013 clusters were subjected to whole-mount in situ hybridization to analyze the gene expression profile in the tadpole larva. A total of 361 clusters showed expression specific to a certain tissue or organ: 96 showed epidermis-specific expression, 60 were specific to the nervous system, 108 to endoderm, 34 to mesenchyme, 5 to trunk lateral cells, 4 to trunk ventral cells, 23 to notochord, 28 to muscle, and 3 to siphon rudiments. In addition, 190 clusters showed expression in multiple tissues. Moreover, nervous system-specific genes showed intriguing expression patterns dependent on the cluster. The present study highlights a broad spectrum of genes that are used in the formation of one of the most primitive chordate body plans as well as for the function of various types of tissue and organ and also provides molecular markers for individual tissues and organs constituting the Ciona larva. (C) 2002 Elsevier Science (USA).

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  • 1D0915 Visual cycle genes in ascidians : insights into the origin of the vertebrate visual cycle

    Nakashima Y., Kusakabe T., Kusakabe R., Horie T., Iwamoto H., Terakita A., Shichida Y., Tsuda M.

    Seibutsu Butsuri   42 ( 2 )   S22   2002

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  • Ci-opsin1, a vertebrate-type opsin gene, expressed in the larval ocellus of the ascidian Ciona intestinalis

    T Kusakabe, R Kusakabe, Kawakami, I, Y Satou, N Satoh, M Tsuda

    FEBS LETTERS   506 ( 1 )   69 - 72   2001.9

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    A novel gene encoding visual pigment, Ci-opsin1, was identified in a primitive chordate, the ascidian, Ciona intestinalis. Molecular phylogenetic analysis and the exon-intron organization suggest that Ci-opsin1 is closely related to the retinal and pineal opsins of vertebrates. During embryogenesis, Ci-opsin1 transcripts were first detected in part of the brain of mid tailbud embryos; its expression was confined to photoreceptor cells of the ocellus (eye spot) in the larval brain as development proceeded. These results suggest a common descent of the ascidian ocellus and the vertebrate eyes. The ocellus of ascidian larvae may represent an ancestral state of the vertebrate eye. (C) 2001 Federation of European Biochemical Societies, Published by Elsevier Science B.V. All rights reserved.

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  • Ci-opsin1, a vertebrate-type opsin gene, expressed in the larval ocellus of the ascidian Ciona intestinalis Reviewed

    T Kusakabe, R Kusakabe, Kawakami, I, Y Satou, N Satoh, M Tsuda

    FEBS LETTERS   506 ( 1 )   69 - 72   2001.9

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    A novel gene encoding visual pigment, Ci-opsin1, was identified in a primitive chordate, the ascidian, Ciona intestinalis. Molecular phylogenetic analysis and the exon-intron organization suggest that Ci-opsin1 is closely related to the retinal and pineal opsins of vertebrates. During embryogenesis, Ci-opsin1 transcripts were first detected in part of the brain of mid tailbud embryos; its expression was confined to photoreceptor cells of the ocellus (eye spot) in the larval brain as development proceeded. These results suggest a common descent of the ascidian ocellus and the vertebrate eyes. The ocellus of ascidian larvae may represent an ancestral state of the vertebrate eye. (C) 2001 Federation of European Biochemical Societies, Published by Elsevier Science B.V. All rights reserved.

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  • O-Glycosylation of G-protein-coupled receptor, octopus rhodopsin - Direct analysis by FAB mass spectrometry Reviewed

    M Nakagawa, T Miyamoto, R Kusakabe, S Takasaki, T Takao, Y Shichida, M Tsuda

    FEBS LETTERS   496 ( 1 )   19 - 24   2001.5

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    In addition to the N-glycan that is evidently conserved in G-protein-coupled receptors (GPCRs), O-glycans in the N-terminus of GPCRs have been suggested. Using a combination of enzymatic and manual Edman degradation in conjunction with G-protein coupled receptor mass spectrometry, the structure and sites of O-glycans in octopus rhodopsin are determined. Two N-acetylgalactosamine residues are O-linked to Thr4 and Thr5 in the N-terminus of octopus rhodopsin, Further, we found chicken iodopsin, but not bovine rhodopsin, contains N-acetylgalactosamine. This is the first direct evidence to determine the structure and sites of O-glycans in GPCRs, (C) 2001 Published by Elsevier Science B,V, on behalf of the Federation of European Biochemical Societies.

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  • Why vertebrate visual cell is hyperpolarized?

    Tsuda M., Kawakami I., Kusakabe R., Kusakabe T.

    Seibutsu Butsuri   41   S67   2001

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  • Primary structure and expression patterns of Ci-RGR1, a novel GPCR of the ascidian Ciona intestinalis

    Nakashima Y., Kusakabe R., Kusakabe T., Tuda M.

    Seibutsu Butsuri   41   S69   2001

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  • In vivo analysis of two striated muscle actin promoters reveals combinations of multiple regulatory modules required for skeletal and cardiac muscle-specific gene expression Reviewed

    R Kusakabe, T Kusakabe, N Suzuki

    INTERNATIONAL JOURNAL OF DEVELOPMENTAL BIOLOGY   43 ( 6 )   541 - 554   1999.9

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    We isolated two striated muscle actin genes from medaka Oryzias latipes. OIMA1 is a skeletal muscle actin gene expressed in semitic muscle and head muscle and OIMA2 is probably a cardiac muscle actin gene expressed in both semitic and cardiac muscle. The differential transcription mechanisms for these two genes were examined in embryos by introducing fusion genes in which the OIMA1 or OIMA2 upstream region was connected to the green fluorescent protein gene. Embryos were injected with these fusion genes at the 2-cell stage. A fusion gene containing the region up to -949 of OIMA1 exhibited strong expression in semitic muscle. The coexistence of two regions, -949/-662 and -421/-201, is necessary for skeletal muscle specific expression of OIMA1. Two E boxes and other unidentified sequences cooperatively function to achieve the full activity of the enhancer -949/-662. As for OIMA2 the region up to -520 is sufficient for strong muscle-specific expression. The region between -520 and -174 of OIMA2 is necessary for specific expression in both skeletal and cardiac muscles. In addition to the CArG box located at -140, an E-box at -430 is important for the expression in cardiac muscle as well as skeletal muscle, When the enhancers for the two muscle actin genes were switched and combined with each other's promoter, they were able to upregulate tissue-specific expression according to their origin. These results suggest that distinct expression patterns of OIMA1 and OIMA2 are regulated by combination of regulatory modules, each of which contains multiple regulatory elements.

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  • Genomic organization and evolution of actin genes in the amphioxus Branchiostoma belcheri and Branchiostoma floridae Reviewed

    R Kusakabe, N Satoh, LZ Holland, T Kusakabe

    GENE   227 ( 1 )   1 - 10   1999.2

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    We previously described the cDNA. cloning and expression patterns of actin genes from amphioxus Branchiostoma floridae (Kusakabe, R., Kusakabe, T., Satoh, N., Holland, N.D., Holland, L.Z., 1997. Differential gene expression and intracellular mRNA localization of amphioxus actin isoforms throughout development: implications for conserved mechanisms of chordate development. Dev. Genes Evol. 207, 203-215). In the present paper, we report the characterization of cDNA clones for actin genes from a closely related species, Branchiostoma belcheri, and the exon-intron organization of B. floridae actin genes. Each of these two amphioxus species has two types of actin genes, muscle and cytoplasmic. The coding and non-coding regions of each type are well-conserved between the two species. A comparison of nucleotide sequences of muscle actin genes between the two species suggests that a gene conversion may have occurred between two B. floridae muscle actin genes BfMA1 and BfMA2. From the conserved positions of introns between actin genes of amphioxus and those of other deuterostomes, the evolution of deuterostome actin genes can be inferred. Thus, the presence of an intron at codon 328/329 in Vertebrate muscle and cytoplasmic actin genes but not in any known actin gene in other deuterostomes suggests that a gene conversion may have occurred between muscle and cytoplasmic actin genes during the early evolution of the vertebrates after separation from other deuterostomes. A Southern blot analysis of genomic DNA revealed that the amphioxus genome contains multiple muscle and cytoplasmic actin genes. Some of these actin genes seem to have arisen from recent duplication and gene conversion. Our findings suggest that the multiple genes encoding muscle and cytoplasmic actin isoforms arose independently in each of the three chordate lineages and that gene duplications and gene conversions established the extant actin multigene family during the evolution of chordates. (C) 1999 Elsevier Science B.V. All rights reserved.

    DOI: 10.1016/S0378-1119(98)00608-8

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Books

  • Evolutionary Developmental Biology - A Reference Guide

    ( Role: Contributor)

    Springer  2020 

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  • 動物学の百科事典

    公益社団法人日本動物学会

    丸善出版  2018.9  ( ISBN:4621303090

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    Total pages:800  

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  • 岩波 生物学辞典 第5版

    巌佐 庸, 倉谷 滋, 斎藤 成也, 塚谷 裕一

    岩波書店  2013.2  ( ISBN:9784000803144

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    Total pages:2192  

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  • ウォルパート発生生物学

    武田洋幸, 田村宏治

    メディカルサイエンスインターナショナル  2012.10  ( ISBN:4895927164

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    Total pages:672  

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  • Medaka : biology, management, and experimental protocols

    木下 政人

    Wiley-Blackwell  2009  ( ISBN:9780813808710

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    Language:English  

    CiNii Books

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  • 動物の形態進化のメカニズム (シリーズ 21世紀の動物科学)

    倉谷 滋, 佐藤 矩行

    培風館  2007.7  ( ISBN:456308283X

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    Total pages:244  

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MISC

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Research Projects

  • 魚類ゲノムの比較解析による運動器形成機構の解明

    2025.4 - 2026.3

    公益財団法人三菱財団  自然科学研究助成  一般助成

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  • 運動器の形成・維持・老化における間葉系間質細胞の機能

    2024.4 - 2025.3

    公益財団法人ノバルティス科学振興財団  第37回ノバルティス研究奨励金 

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    Authorship:Principal investigator 

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  • 脊椎動物の「首」をつくりだす遺伝子機能の解明

    Grant number:22K06243  2022.4 - 2025.3

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    日下部 りえ

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    Grant amount:\4160000 ( Direct Cost: \3200000 、 Indirect Cost:\960000 )

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  • 鰓下筋群の複雑化をもたらした新規の遺伝子制御

    2019.4 - 2021.3

    文部科学省  科学研究費補助金(基盤研究(C)) 

    日下部 りえ

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    Authorship:Principal investigator  Grant type:Competitive

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  • 第二心臓予定領域の進化

    2016.4 - 2018.3

    文部科学省  科学研究費補助金(基盤研究(C)) 

    日下部 りえ

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\4940000 ( Direct Cost: \3800000 、 Indirect Cost:\1140000 )

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  • Evolutionary developmental analysis for the origin and migration of primordial germ cells

    Grant number:25440108  2013 - 2015

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    Kusakabe Rie, INOUE Kunio

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    Grant amount:\5330000 ( Direct Cost: \4100000 、 Indirect Cost:\1230000 )

    The cyclostome lampreys, a group of vertebrates that diverged early in evolution, retain various ancestral morphological features. In order to clarify the evolutionary pathway of origin and developmental behavior of undifferentiated precursor cells, including primordial germ cells, we conducted examination of expression patterns of developmental genes and histological observation. We also attempted the new experimental techniques for lineage tracing after introduction of artificial gene constructs. Comparison with insights from other vertebrate species implied the evolutionary relationships of the migratory undifferentiated precursor cells and the establishment of complex and diverse morphology of vertebrates.

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  • Developmental mechanisms of primordial germ cells in the cyclostome lamprey

    Grant number:23770250  2011 - 2012

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Young Scientists (B)

    KUSAKABE Rie

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    Grant amount:\4550000 ( Direct Cost: \3500000 、 Indirect Cost:\1050000 )

    The cyclostome lampreys are a group of vertebrates that diverged early in evolution and retain various ancestral morphological traits. In order to explore the developmental mechanisms underlying the establishment of germ cell lineage, we examined the emergence of primordial germ cells (PGCs) and the gonads in the embryonic lampreys. Genes involved in the specification of PGCs, migration of PGCs to the gonads and in the establishment of gonadal primordium were newly identified by RT-PCR. Expression of each gene was analyzed by whole mount in situ hybridization. The results showed that maternal mRNAs of several germ cell-related genes were localized in fertilized eggs of the lamprey. We also established the experimental system in which migratory cells in the early embryos can be traced with the expression of the reporter fluorescent protein driven by upstream regulatory sequence of CXCR4 gene.

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  • microRNAの多様性と脊椎動物の系統進化

    Grant number:21770257  2009 - 2010

    文部科学省  科学研究費補助金(若手研究(B))  若手研究(B)

    日下部 りえ

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    Grant amount:\4550000 ( Direct Cost: \3500000 、 Indirect Cost:\1050000 )

    脊椎動物の各系統での小分子機能性RNAの重複の度合いや機能の変遷を解明するため、複数の動物でmicroRNAの同定と機能解析を行った。平成22年度はまず、尾索動物ホヤで筋肉細胞特異的に発現するmiR-1, miR-133について、発現パターンと転写制御機構を解析した。ノザンブロット実験により各成熟型miRの発現を検出したところ、1)miR-133は変態前の胚発生過程において発現するが、miR-1は発現しないこと、2)成体では、miR-133は体壁筋のみに、miR-1は体壁筋と心筋の両方に発現することが明らかになった。さらに、miR-1/miR-133は単一の一次産物として、尾芽胚の筋肉細胞の核に局在し、その発現はmiR-1の上流領域に制御されることが見出された。現在・真骨魚類メダカについても前駆体型から成熟型へのプロセシングについて解析中である。またヤツメウナギに関し、一個のmiR-1と二個のmiR-133の相同遺伝子を同定し、前駆体の二次構造を予測した。成熟型miR-133についてノザンブロット解析を行ったところ、筋肉の発生過程で発現していた。以上の知見から、1)miR-1/miR-133は脊索動物の共通祖先においてすでに、遺伝子対としてゲノム上の近傍に位置するようになり、一続きの前駆体として筋肉特異的に産生されるようになったこと、2)脊索動物の共通祖先においてこの遺伝子対...

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  • メダカ突然変異体を用いた骨格筋パターン形成機構の進化に関する研究

    Grant number:18770212  2006 - 2008

    文部科学省  科学研究費補助金(若手研究(B))  若手研究(B)

    日下部 りえ

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    Grant amount:\3700000 ( Direct Cost: \3700000 )

    平成19年度は、骨格筋パターニング機構をメダカとヤツメ ウナギで比較する目的で、生きたヤツメウナギ胚を用いた実験を行った。まず、ヤツメウナギの側板中胚葉であると考えられる領墟をDiIでラベルし、アンモシーテス幼生期まで飼育し固定した。次に胚を凍結切片にし、筋肉をMF20抗体で染色して観察した。その結果、DiIセラベルされた側板中胚葉と体節由来の筋肉は細胞が混じり合わないことが分かった。顎口類の移動性軸下筋では体節由来の筋芽細胞が側板中胚葉の環境下で分化するが、今回の実験からはヤツメウナギではそのような細胞間相互作用が起こらないという可能性が示唆された。また、皮筋節の発生に必須であると予想されるPax3/7遺伝子に対するモリフォリノオリゴヌレオチドを合成し、受精卵に顕微注入して発生させた。モリフォリノを注入された胚の多くに、頭部が前方に伸長できないという表現型がみられた。メダカでは、筋肉特異的microRNAであるmiR-1,miR-206,miR-133に対しLNA(locked-nucleic acid-modified oligonucleotide)プローブを作製し、in situハイブリダイゼーションを行って発現領域を同定した。その結果、miR-206はメダカ胚においてすべての骨格筋に発現するのに対し、miR-1は体幹部の筋肉には発現するが、移動性軸下筋である対鰭の...

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  • メダカ突然変異体Daを用いた脊椎動物筋肉発生様式に関する研究

    2004 - 2007

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  • 脊椎動物骨格筋の背腹パターン形成機構とその進化に関する研究

    Grant number:16770174  2004 - 2005

    文部科学省  科学研究費補助金(若手研究(B))  若手研究(B)

    日下部 りえ

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\3100000 ( Direct Cost: \3100000 )

    平成17年度は、前年度に確立した骨格筋可視化メダカ系統(野生型、Da変異体)を用いて、Daの原因遺伝子であるZic1の強制発現実験を行うとともに、in situハイブリダイゼーションを用いて初期発生に働く遺伝子の発現パターンを調べた。硬節(sclerotome)のマーカーであるtwistは、野生型では体節の腹側で発現が始まり背側へと発現領域を広げるが、Da変異体では次第に発現が弱まってしまい、背側で発現することはなかった。また、Shh(ソニックヘッジホッグ)カスケードの阻害剤であるシクロパミンで野生型あるいはDa胚を処理すると、筋節の発生が異常になり、水平筋中隔が欠失、すなわち軸上・軸下の境界が失われた。しかしどちらの系統においても、対鰭には正常な筋芽細胞の移動が見られた。すなわち、対鰭に移動する特殊化した軸下筋は、軸上・軸下の区別がない状態でもアイデンティティを獲得できることがわかった。軸上・軸下の区別を持たない無顎類ヤツメウナギからZic遺伝子群を含む複数の筋肉発生関連遺伝子を単離し発現パターンを解析した。ヤツメウナギZic遺伝子は前脳などの中枢神経系に強く発現しているほか、体節の背側にも発現が見られた。また顎口類において軸下筋の発生に重要なPax3遺伝子、およびPax3に制御され四肢などに移動する軸下筋のグループのマーカーとして知られるLbx1遺伝子は、ヤツメウナギの体...

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  • 円口類ヤツメウナギを用いた脊椎動物骨格筋形態の進化に関する研究

    2000 - 2007

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