変異型SYCP3が相同染色体分配に与えるインパクトと習慣流産

2011 Fiscal Year Annual Research Report

• Project Area: The germline: its developmental cycle and epigenome network
• Project/Area Number: 23013019
• Research Institution: Fujita Health University
• Principal Investigator: 倉橋 浩樹 藤田保健衛生大学, 総合医科学研究所, 教授 (30243215)
• Keywords: 異数体 / 習慣流産 / SYCP3 / 対合 / 卵巣 / 器官培養 / 減数分裂 / 遺伝子導入

Research Abstract

わたしたちは、異数性による流産を繰り返していると思われる習慣流産の3名の患者で、第1減数分裂時の相同染色体の対合に関与するシナプトネマ複合体の蛋白をコードするSYCP3遺伝子の変異を同定した。これらの変異はすべてスプライシング部位の変異で、第8イントロンの残存、もしくは、第8エクソンのスキップをきたし、C末端の変異型SYCP蛋白を産生する。これまでの研究で、これらのC末端の変異蛋白が優性阻害効果によって、正常のSYCP3蛋白のファイバー形成を阻害することを、細胞株を用いた実験で証明している。しかし、実際の減数分裂時の卵細胞における変異型SYCP3蛋白の挙動、染色体分配に対する影響に関しては未解明である。そこで、本研究では、これら変異型SYCP3蛋白をマウスの卵細胞に強制発現させ、減数分裂細胞レベルでの異常の検出を試み、習慣流産の発症メカニズムに迫ることを目的としている。本年度は、マウス卵巣の器官培養の実験系を確立し、培養条件などの至適化に成功した。また、GFP遺伝子や正常SYCP3蛋白の発現ベクターを電気穿孔法により導入すること、ユビキタスプロモーターや減数分裂前期特異的プロモーターのもとに発現させること、タグの抗体を用いて発現蛋白を内因性蛋白と区別して検出することに成功し、実験系を確立することができた。強制発現させた正常SYCP3蛋白は染色体の軸上に同定され、内因性のSYCP3蛋白とともにシナプトネマ複合体を形成することが観察できた。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

当初、1年目は実験系の確立に、2年目は実際の変異蛋白の挙動の観察にあてる計画を立てていたが、本年度で卵巣の器官培養による実験系の確立、条件の至適化ができた、

Strategy for Future Research Activity

次年度は、予定通り、習慣流産患者で同定された遺伝子変異を持つ発現ベクターを卵細胞に導入して、その表現型の変化を異数体の発生という観点から観察し、習慣流産の発生メカニズムに迫る予定である。

Research Products

• [Journal Article] Mechanism of complex gross chromosomal rearrangements : A commentary on Concomitant microduplications of MECP2 and ATRX in male patients with severe mental retardation (2012)
  • Author(s): Kurahashi H, Ohye T, Inagaki H, Kogo H, Tsutsumi M
  • Journal Title: J Hum Genet Volume: 57 Pages: 81-83
  • DOI: 10.1038/jhg.2011.143
• [Journal Article] Molecular basis of maternal age-related increase in oocyte aneuploidy (2012)
  • Author(s): Kurahashi H, Tsutsumi M, Nishiyama S, Kogo H, Inagaki H, Ohye T
  • Journal Title: Congenit Anom (Kyoto) Volume: 52 Pages: 8-15
  • DOI: 10.1111/j.1741-4520.2011.00350.x
  • Peer Reviewed
• [Journal Article] Characterization of a novel mouse gene encoding an SYCP3-like protein that re-localizes from the XY body to the nucleolus during prophase of male meiosis I (2011)
  • Author(s): Tsutsumi M, Kogo H, Kowa-Sugiyama H, Inagaki H, Ohye T, Kurahashi H
  • Journal Title: Biol Reprod Volume: 85 Pages: 165-171
  • DOI: 10.1095/biolreprod.110.087270
  • Peer Reviewed
• [Journal Article] Pathogenic exon-trapping by SVA retrotransposon and rescue in Fukuyama muscular dystrophy (2011)
  • Author(s): Taniguchi-Ikeda M, Kobayashi K, Kanagawa M, Yu CC, Mori K, Oda T, Kuga A, Kurahashi H, Akman HO, DiMauro S, Kaji R, Yokota T, Takeda S, Toda T
  • Journal Title: Nature Volume: 478 Pages: 127-131
  • DOI: 10.1038/nature10456
  • Peer Reviewed
• [Journal Article] A rare synaptonemal complex protein 3 gene variant in unexplained female infertility (2011)
  • Author(s): Nishiyama S, Kishi T, Kato T, Suzuki M, Bolor H, Udagawa Y, Kurahashi H
  • Journal Title: Mol Hum Reprod Volume: 17 Pages: 266-271
  • DOI: 10.1093/molehr/gaq098
  • Peer Reviewed
• [Journal Article] Polymorphism in annexin A5 gene promoter in Japanese women with recurrent pregnancy loss (2011)
  • Author(s): Miyamura H, Nishizawa H, Ota S, Suzuki M, Inagaki A, Egusa H, Nishiyama S, Kato T, Pryor-Koishi K, Nakanishi I, Fujita T, Imayoshi Y, Markoff A, Yanagihara I, Udagawa Y, Kurahashi H
  • Journal Title: Mol Hum Reprod Volume: 17 Pages: 447-452
  • DOI: 10.1093/molehr/gar008
  • Peer Reviewed
• [Journal Article] DNA secondary structure is influenced by genetic variation and alters susceptibility to de novo translocation (2011)
  • Author(s): Kato T, Inagaki H, Tong M, Kogo H, Ohye T, Yamada K, Tsutsumi M, Emanuel BS, Kurahashi H
  • Journal Title: Mol Cytogenet Volume: 4 Pages: 18
  • DOI: 10.1186/1755-8166-4-18
  • Peer Reviewed
• [Journal Article] Comparative gene expression profiling of placentas from patients with severe pre-eclampsia and unexplained fetal growth restriction (2011)
  • Author(s): Nishizawa H, Ota S, Suzuki M, Kato T, Sekiya T, Kurahashi H, Udagawa Y
  • Journal Title: Reprod Biol Endocrinol Volume: 9 Pages: 107
  • DOI: 10.1186/1477-7827-9-107
  • Peer Reviewed
• [Journal Article] Impact of indoleamine 2,3-dioxygenase on the antioxidant system in the placentas of severely pre-eclamptic patients (2011)
  • Author(s): Nishizawa H, Suzuki M, Pryor-Koishi K, Sekiya T, Tada S, Kurahashi H, Udagawa Y
  • Journal Title: Syst Biol Reprod Med Volume: 57 Pages: 174-178
  • DOI: 10.3109/19396368.2011.587590
  • Peer Reviewed
• [Journal Article] Global gene expression profiling in early-stage polycystic kidney disease in the Han : SPRD Cy rat identifies a role for RXR signaling (2011)
  • Author(s): Kugita M, Nishii K, Morita M, Yoshihara D, Kowa-Sugiyama H, Yamada K, Yamaguchi T, Wallace DP, Calvet JP, Kurahashi H, Nagao S
  • Journal Title: Am J Physiol Renal Physiol Volume: 300 Pages: F177-188
  • DOI: 10.1152/ajprenal.00470.2010
  • Peer Reviewed
• [Journal Article] PPAR-{gamma} agonist ameliorates kidney and liver disease in an orthologous rat model of human autosomal recessive polycystic kidney disease (2011)
  • Author(s): Yoshihara D, Kurahashi H, Morita M, Kugita M, Hiki Y, Aukema HM, Yamaguchi T, Calvet JP, Wallace DP, Nagao S
  • Journal Title: Am J Physiol Renal Physiol Volume: 300 Pages: F465-474
  • DOI: 10.1152/ajprenal.00460.2010
  • Peer Reviewed
• [Journal Article] Lack of Association between Orexin Receptor Genes Polymorphisms and Obstructive Sleep Apnea Syndrome in Japanese (2011)
  • Author(s): Ahmed WA, Mori T, Nishimura Y, Kitanaka T, Kato T, Bhardwaj KA, Kurahashi H, Suzuki K
  • Journal Title: Sleep Biol Rhythms Volume: 9 Pages: 73-77
  • Peer Reviewed
• [Journal Article] Chromosomal translocations and palindromic AT-rich repeats
  • Author(s): Kato T, Kurahashi H, Emanuel BS
  • Journal Title: Curr Opin Genet Dev Volume: (In press)
  • Peer Reviewed
• [Journal Article] HORMAD1-dependent checkpoint/surveillance mechanism eliminates asynaptic oocytes
  • Author(s): Kogo H, Tsutsumi M, Ohye T, Inagaki H, Abe T, Kurahashi H
  • Journal Title: Genes Cells Volume: (In press)
  • Peer Reviewed
• [Journal Article] A Functional variation in the hypocretin neuropeptide precursor gene may be associated with obstructive sleep apnea syndrome in Japan
  • Author(s): Ahmed WA, Tsutsumi M, Nakata S, Mori T, Nishimura Y, Fujisawa T, Kato I, Nakashima M, Kurahashi H, Suzuki K
  • Journal Title: Laryngoscope Volume: (In press)
  • Peer Reviewed
• [Journal Article] Serum neutrophil gelatinase-associated lipocalin during the early postoperative period predicts the recovery of graft function after kidney transplantation from donors after cardiac death
  • Author(s): Kusaka M, Iwamatsu F, Kuroyanagi Y, Nakaya M, Ichino M, Marubashi S, Nagano H, Shiroki R, Kurahashi H, Hoshinaga K
  • Journal Title: J Urol Volume: (In press)
  • Peer Reviewed
• [Presentation] Global gene expression profiling in kidneys of PPAR-gamma agonist-treated PCK rats, an orthologous model of human ARPKD (2011)
  • Author(s): Yoshihara D, Kugita M, Kurahashi H, Morita M, Hiki Y, Yamaguchi T, Aukema HM, Wallace DP, Calvet JP, Toyohara T, Abe T, Nagao S
  • Organizer: American Society of Nephrology 44th Annual Meeting & Scientific Exposition
  • Place of Presentation: Philadelphia, PA USA
  • Year and Date: 20111100
• [Presentation] Mechanism of recurrent translocation t(11;22) initiated by cruciform conformation of palindromic sequences (2011)
  • Author(s): Inagaki H, Ohye T, Kogo H, Tsutsumi M, Kato T, Tong M, Emanuel BS, Kurahashi H
  • Organizer: The 12th International Congress of Human Genetics
  • Place of Presentation: Montreal, Canada
  • Year and Date: 20111011-20111015
• [Presentation] Identification of a recombination hotspot sequence at the breakpoint region of the 22q11 deletion using a yeast model (2011)
  • Author(s): Ohye T, Inagaki H, Kogo H, Tsutsumi M, Emanuel BS, Kurahashi H
  • Organizer: The 12th International Congress of Human Genetics
  • Place of Presentation: Montreal, Canada
  • Year and Date: 20111011-20111015
• [Presentation] HORMAD1-deficiency causes azoospermia in males and pregnancy loss in females (2011)
  • Author(s): Kogo H, Tsutsumi M, Ohye T, Inagaki H, Kurahashi H
  • Organizer: The 12th International Congress of Human Genetics
  • Place of Presentation: Montreal, Canada
  • Year and Date: 20111011-20111015
• [Presentation] Identification of a novel palindrome mediated translocation associated with the t(3;8) of hereditary renal cancer (2011)
  • Author(s): Kato T, Sheridan MB, Hacker AM, Inagaki H, Glover TW, Plon SE, Drabkin HA, Gemmill RM, Kurahashi H, Emanuel BS
  • Organizer: The 12th International Congress of Human Genetics
  • Place of Presentation: Montreal, Canada
  • Year and Date: 20111011-20111015
• [Presentation] Mutation in the SYCP3 gene identified in a woman with recurrent pregnancy loss affect the synaptonemal complex conformation at meiotic prophase I (2011)
  • Author(s): Tsutsumi M, Kogo H, Inagaki H, Ohye T, Kurahashi H
  • Organizer: The 12th International Congress of Human Genetics
  • Place of Presentation: Montreal, Canada
  • Year and Date: 20111011-20111015
• [Presentation] HORMAD1-dependent synapsis checkpoint in mammalian meiosis (2011)
  • Author(s): Kogo H, Tsutsumi M, Ohye T, Inagaki H, Kurahashi H
  • Organizer: 2011 FASEB Summer Research Conferences, Genetic Recombination & Genome Rearrangements
  • Place of Presentation: Steamboat Springs, USA
  • Year and Date: 20110724-20110729
• [Presentation] Polymorphisms in the annexin A5 gene promoter in Japanese women with recurrent pregnancy loss (2011)
  • Author(s): Miyamura H, Nishizawa H, Ota S, Suzuki M, Inagaki A, Egusa H, Nishiyama S, Kato T, Pryor-Koishi K, Nakanishi I, Fujita T, Imayoshi Y, Markoff A, Yanagihara I, Udagawa Y, Kurahashi H
  • Organizer: 27th Annual Meeting of ESHRE-Stockholm 2011
  • Place of Presentation: Stockholm, Sweden
  • Year and Date: 20110703-20110706
• [Presentation] Polymorphisms in the annexin A5 gene promoter in Japanese women with recurrent pregnancy loss (2011)
  • Author(s): Kurahashi H, Miyamura H, Nishizawa H, Ota S, Suzuki M, Inagaki A, Egusa H, Nishiyama S, Kato T, Pryor-Koishi K, Nakanishi I, Fujita T, Imayoshi Y, Negi R, Markoff A, Yanagihara I, Udagawa, Y
  • Organizer: European Society of Human Genetics Conference 2011.
  • Place of Presentation: Amsterdam, Netherlands
  • Year and Date: 20110528-20110531
• [Presentation] Polymorphisms in the annexin A5 gene promoter in Japanese women with recurrent pregnancy loss (2011)
  • Author(s): Miyamura H, Nishizawa H, Ota S, Suzuki M, Inagaki A, Egusa H, Nishiyama S, Kato T, Pryor-Koishi K, Nakanishi I, Fujita T, Imayoshi Y, Markoff A, Yanagihara I, Udagawa Y, Kurahashi H
  • Organizer: 59th Annual Clinical Meeting, ACOG
  • Place of Presentation: Washington DC, USA
  • Year and Date: 20110430-20110504
• [Presentation] PPAR-gamma agonist ameliorates kidney and liver disease in an orthologous rat model of human autosomal recessive polycystic kidney disease (2011)
  • Author(s): Yoshihara D, Kurahashi H, Morita M, Kugita M, Hiki Y, Aukema HM, Yamaguchi T, Calvet JP, Wallace DP, Nagao S
  • Organizer: World Congress of Nephrology 2011
  • Place of Presentation: Vancouver, Canada
  • Year and Date: 20110400
• [Remarks]
  • URL: http://www.fujita-hu.ac.jp/ICMS/
• [Remarks] 分子遺伝学研究部門
  • URL: http://www.fujita-hu.ac.jp/~genome/mg/