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2002 Fiscal Year Final Research Report Summary

Studies on apyrene function and its application for sperm storage in lepidopteran insects

Research Project

Project/Area Number 13556009
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 蚕糸・昆虫利用学
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

KEN Sahara  Hokkaido Univ., Grad. School of Agr., Assistant Prof., 大学院・農学研究科, 助手 (30241368)

Co-Investigator(Kenkyū-buntansha) BANNO Yutaka  Kyushu Univ. Grad. School of Agr., Assoc. Prof., 大学院・農学研究科, 助教授 (50192711)
SAITO Yutaka  Hokkaido Univ., Grad. School of Agr., Prof., 大学院・農学研究科, 教授 (20142698)
BANDO Hisanori  Hokkaido Univ., Grad. School of Agr., Prof., 大学院・農学研究科, 教授 (20189731)
TAKEMURA Yoko  Inst. Silkworm Genet. Breed., Research Scientist, 蚕業技術研究所, 研究員
YAMASHIKI Naoko  Rakuno-Gakuen Univ., Biology Lab., Assoc. Prof., 環境システム学部, 助教授 (40112578)
Project Period (FY) 2001 – 2002
KeywordsBombyx mori / eypyrne sperm / apyrene spem / cryopreservation / artificial insemination / dimorphic sperm / maintenance of bioresorce / double copulation
Research Abstract

Lepidopteran insects have dimorphic sperm system, fertilizing eupyrene sperm and anucleate apyrene sperm. We found that polyploid male of Bombyx mori have only functional eupyrene. On the other hand the apyrene sperm is produced alone by heat-shock treatment to pupae of Daizo strain in the silkworm. Thus we successfully separated the eupyrene sperm from the apyrene sperm. Double copulation and artificial insemination experiments with mixing of the separated sperm proved that the apyrene sperm is inevitable for successful fertilization by the eupyrene sperm. These results are first experimental proof of the function of apyrene sperm concerning the fertilization. This experimental system enables us to observe the behavior of individual sperm populations in female reproductive organs. The eupyrene sperm cannot proceed to spermatheca without mixing with the apyrene sperm, resulting no fertilization success.
Artificial insemination to females with cryopreserved eupyrene sperm resulted normal fertility if intact apyrene sperm was supplied. On the other hand cryopreserved apyrene sperm sometimes lost their ability after freeze-and-thaw treatment. Decrease in fertility of cryopreserved sperm seems to be mainly caused by sever damage of apyrene sperm. This greatly varies among silkworm strains. Utilization of resistant strains against freeze-and-thaw for producing polyploid males may be one of the ways to increase the stability of apyrene sperm function. We improved the fertility of cryopreseved sperm stock by applying the apyrene sperm which was kept in liquid nitrogen. The combination system of artificial insemination technique, cryopreservation of sperm and application of separated apyrene sperm constructed in this study is basis to develop the new method to maintain the genetic resources of the silkworm.

  • Research Products

    (63 results)

All Other

All Publications (63 results)

  • [Publications] Sahara K: "Application of artificial insemination technique to separated eupyrene and apyrene sperm in Bombvx mori"Journal of Experimental Zoology. (in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sahara K: "Roles of actin networks in peristaltic squeezing of sperm bundles in Bombyx mori"Journal of Morphology. (in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawamura N: "Glucose and ecdysteroid increase apyrene sperm production in in vitro cultivation of spermatocysts of Bombyx mori"Journal of Insect Physiology. (in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sahara K: "Moth sex chromatin probed by comparative genomic hybridization (CGH)"Genome. (in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Matsuyama T: "Functional analysis of an immediate early gene, iel, of Bombyx mori nucleopolyhedrovirus in mammalian cells"Journal of Insect Biotechnology and Sericology. (in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamagishi J: "DNA Microarrays of baculovirus genomes ; Differential expression of viral genes in two susceptible insect cell lines"Archives of Virology. (in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 小島 桂: "トランスジェニックカイコの家蚕核多角体病ウイルスに対する抵抗性"北海道大学北方圏フィールド科学センター研究農場報告. (印刷中). (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Saito Y: "Differences in diapause attributes between two forms distinguished by male-to-male aggression in a subsocial spider mite, Schizotetranychus miscanthi"Ecological Research. 17・6. 645-653 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawamura N: "In vitro cultivation of spermatocysts to matured sperm in the silkworm, Bombyx mori"Development, Growth and Differentiation. 44・4. 273-280 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sahara K: "Double copulation of a female with sterile diploid and polyploid males recovers fertility in Bombyx mori"Zygote. 10・1. 23-29 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Isobe R: "Antisense and double-strand RAN interferenece in a silkworm ovarian cell line"Journal of Insect Biotechnology and Sericology. 71・1. 43-47 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ito T: "Cloning and expression of novel crystal protein genes cry39A and 39orf from Bacillus thuringiensis subsp. aizawai BUN1-14 encoding mosquitocidal proteins"Journal of Insect Biotechnology and Sericology. 71・3. 123-128 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamada Y: "Use of an N-terminal half truncated IE1 as an antagonist of IE1, an essential regulatory protein in baculovirus"Virus Research. 90・1-2. 253-261 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamashiki N: "Presence of Phenol DP-glucuronosyltransferase in bovine alveolar macrophages and bronchial epithelial cells"Toxicology. 176・3. 221-227 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yukuhiro K: "Significant Levels of Sequence Divergence and Gene Rearrangements have Occurred Between the Mitochondrial Genomes of the Wild Mulberry Silkmoth, Bombyx mandarina, and its Close Relative, the Domesticated Silkmoth, Bombyx mori"Molecular Biology and Evolution. 19・8. 1385-1389 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Lin J-Z: "A new species of Tarsonemus (Acari : Tarsonemidae),prey on eggs of Schizotetranychus longus Saito. J. Acarol. Soc. Jpn. 11, 85-89"Journal of Acarology Society Japan. 11・1. 85-89 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Marec F: "Meiotic pairing of sex chromosome fragments and its relation to atypical transmission of a sex-linked marker in Ephestia kuehniella (Insecta, Lepidoptera)"Heredity. 87・12. 659-671 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 小島 桂: "家蚕濃核病ウイルス(BmDNV-2)特異的DNAポリメラーゼ活性と末端配列認識タンパク質の解析"日本蚕糸学雑誌. 70・2. 103-108 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 佐原 健: "天蚕全齢人工飼料育に用いる飼料樹葉特性の検討"日本応用動物昆虫学雑誌. 45・2. 94-98 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawamura N: "Meiotic pairing of sex chromosome fragments and its relation to atypical Significance of peristaltic squeezing of sperm bundles in the silkworm, Bombyx mori : Elimination of irregular eupyrene sperm nuclei of the triploid"Zygote. 9・2. 159-166 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Banno Y: "Inheritance of one cocoon shape mutant named "Flossycocoon" of the silkworm, Bombyx mori"Journal of Insect Biotechnology and Sericology. 70・3. 189-192 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakamura T: "Genetics of the "wild silkworm translucent" mutant (ows), discovered in the progenies after the cross between the domesticated silkworm, Bombyx mori and the wild mulberry silkworm, Bombyxmandarina"International Journal of Wild Silkmoth & Silk. 6. 7-10 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Xia QY: "Identification of three annexin IX isoforms generated by alternative splicing of the carboxyl-terminal exon in silkworm, Bombyx mori"Insect Biochemistry and Molecular Biology. 32・1. 9-14 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Xia QY: "Identification of three annexin IX isoforms generated by alternative splicing of the carboxyl-terminal exon in silkworm, Bombyx mori"Insect Biochemistry and Molecular Biology. 32・1. 9-14 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kojima K: "Tandem repetition of baculovirus iel promoter results in upregulation of transcription"Archives of Virology. 146・1. 1-8 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masaki K: "Thermal stability and enzymatic activity of a smaller lysozyme originated from silkworm moth (Bombyx mori)"Journal of Protein Chemistry. 20・2. 107-113 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Koganezawa N: "Construction of an expression system of insect lysozyme lacking thermal stability : the effect of selection of signal sequence on level of expression in the Pichia pastoris expression system"Protein Engineering. 14・9. 705-710 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Li Y: "Genome organization of the densovirus from Bombyx mori (BmDNV-1) and enzyme activity of its capsid"Journal of General Virology. 82・11. 2821-2825 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yanagida H: "Egg depositing behavior as apredetor avoidance tactic of Yezonychus sapporoensis Ehara (Acari, Tetranychidae)"Journal of Ethology. 19・1. 55-62 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Chittenden AR: "Why are there feeding and non-feeding larvae in phytoseiid mites (Acari : Phytoseiidae)?"Journal of Ethology. 19・1. 63-66 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Marec F: "Sex chromosomes in Ephestia kuehniella (Insecta, Lepidoptera) tolerate a telomere loss"Annals de Genetique. 44・Suppl. S131 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawamura N: "A cause of male sterility in polyploid silkworms"Development, Growth and Differentiation. 43・Suppl. S122 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sahara K, Takemura Y: "Application of artificial insemination technique to separated eupyrene and apyrene sperm in Bombyx mori"J Exp Zool.. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sahara K, Marec F, Eickhoff U, Traut W: "Moth sex chromatin probed by comparative genomic hybridization (CGH)"Genome.. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sahara K, Kawamura N: "Roles of actin networks in peristaltic squeezing of sperm bundles in Bombyx mori"J. Morphol.. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawamura N, Sahara K, Fugo H: "Glucose and ecdysteroid increase apyrene sperm production in in vitro cultivation of spermatocysts of Bombyx mori"J. Insect Physiol.. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Matsuyama T, Asano S, Sahara K, Bando H: "Functional analysis of an immediate early gene, ie1, of Bombyx mori nucleopolyhedrovirus in mammalian cells"Journal of Insect Biotechnology and sericology. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamagishi J, Isobe R, Takebuchi T, Bando H: "DNA Microarrays of baculovirus genomes : Differential expression of viral genes in two susceptible insect cell lines"Arch Virol.. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kojima K, Saitoh H, Yamada Y, Tamura T, Isobe R, Asano S, Sahara K, Bando H: "Resistance of transgenic slikworm against Bombyx mori nuclear polyhedrosis virus (BmNPV)"Res Bull Univ Farm Hokkaido Univ.. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Saito Y, Sakagami T, Sahara K: "Differences in diapause attributes between two forms distinguished by male-to-male aggression in a subsocial spider mite, Schizotetranychus miscanthi Saito"Ecol. Res.. 17(6). 645-653 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawamura N, Sahara K: "In vitro cultivation of spermatocysts to matured sperm in the silkworm, Bombyx mori"Dev. Growth Differ.. 44(4). 273-280 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sahara K, Kawamura N: "Double copulation of a female with sterile diploid and polyploid males recovers fertility in Bombyx mori"Zygote. 10(1). 23-29 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Isobe R, Kojima K, Sahara K, Asano S, Bando H: "Antisense and double-strand RAN interference in a silkworm ovarian cell line"J. Insect Biotech. Seric.. 71(1). 43-47 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ito T, Sahara K, Bando H, Asano S: "Cloning and expression of novel crystal protein genes cry39A and 39orf2 from Bacillus thuringiensis subsp.aizawai Bun1-14 encoding mosquitocidal proteins"J Insect Biotech. Seric.. 71. 123-128 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamada Y, Matsuyama T, Quan GX, Kanda T, Tamura T, Sahara K, Asano S, Bando H: "Use of an N-terminal half truncated IE1 as an antagonist of IE1, and essential regulatory protein in baculovirus"Virus Res.. 90(1-2). 253-261 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamashiki N, Yokota H, Sakamoto M, Yuasa A: "Presence of phenol DP-glucuronosy Itransferase in bovine alveolar macrophages and bronchial epithelial cells"Toxicology. 176(3). 221-227 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yukuhiro K, Sezutsu H, Itoh M, Shimizu K, Banno Y: "Significant Levels of Sequence Divergence and Gene Rearrangements have Occurred Between the Mitochondrial Genomes of the Wild Mulberry Silkmoth, Bombyx mandarina, and its Close Relative, the Domesticated Silkmoth, Bombyx mori"Mol. Biol. Evol.. 19(8). 1385-1389 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Lin J-Z., H. Nakao, Y. Saito: "A new species of Tarsonemus (Acari : Tarsonemidase), prey on eggs of Schizotetranychus longus Saito"J. Acarol. Soc. Jpn.. 11. 85-89 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Marec F, Tothov A, Sahara K, Traut W: "Meiotic pairing of sex chromosome fragments and its relation to atypical transmission of a sex-linked marker in Ephestia kuehniella (Insecta, Lepidoptera)"Heredity. 87(12). 659-671 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kojima K, Hirano A, Asano S, Sahara K, Bando H: "Analysis of the BmDNV-2 specific DNA polymerase and the common terminal sequences biding proteins"J Seric Sci Jpn. 70(2). 103-108 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sahara K, Saitoh H, Kikuch K, Yamada Y, Asano S, Bando H: "Comparison of dried-leaf powder of host plants as a component of an artificial diet for the wild silkworm, Antheraea yamamai Guerin-Meneville (Lepidoptera : Saturniieda)"Jpn J Appl Entomol Zool. 45(2). 94-98 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawamura N, Yamashiki N, Saitoh H, Sahara K: "Significance of peristaltic squeezing of sperm bundles in the silkworm, Bombyx mori : Elimination of irregular eupyrene sperm nuclei of the triploid"Zygote. 9(2). 159-166 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Bannno Y, Fujii H, Doira H: "Inheritance of one cocoon shape mutant named "Flossycocoon" of the silkworm, Bombyx mori"Journal of insect biotechnology and sericology. 70. 189-192 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakamura T, Banno Y, Fujii H: "Genetics of the "wild slkworm translucent" mutant(ows), discovered in the progenies after cross between the domesticated silkworm, Bombyx mori and the wild mulberry silkworm, Bombyxmandarina"Int. J. Wild silkmoth & silk. 6. 7-10 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Xia QY, Fujii H, Kusakabe T, Banno Y: "Identification of three annexim IX isoforms generated by alternative splicing of the carboxyl-terminal exon in silkworm, Bombyx mori"Insect Biochem. Mol. Biol.. 32. 9-14 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kojima, K., T. Hayakawa, S. Asano, H. Bando: "Tandem repetition of baculovirus iel promoter results in upregulation of transcription"Archives of Virology. 146. 1-8 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masaki, K., T. Aizawa, N. Koganesawa, T. Nimori, H. Bando, K. Kawano, K. Nitta: "Thermal stability and enzymatic activity of a smaller lysozyme originated from silkworm moth (Bombyx mori)"Journal of Protein Chemistry. 20. 107-113 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Koganezawa N, Aizawa T, Masaki K, Matuura A, Nimori T, Bando H, Kawano K, Nitta K: "Construction of an expression system of insect lysozyme lacking thermal stability : the effect of selection of signal sequence on level of expression in the Pichia pastoris expression system"Protein engineering. 14. 705-710 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Li, Y., Z. Zadori, H. Bando, R. Dubuc, G. Fediere, J. Szeilei, P. Tijssen: "Genome organization of the densovirus from Bombyx mori (BmDNV-1) and enzyme activity of its capsid"J. Gen. Virol.. 82. 2821-2825 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Chittenden AR, Saito Y: "Why are there feeding and non-feeding larvae in phytoseiid mites (A cari : Phytoseiidae)?"J Ethol. 19. 63-66 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yanagida H, Saito Y, Mori K, Chittenden AR: "Egg depositing behavior as apredetor avoidance tactic of Yezonychus sapporoensis Ehara (Acari, Tetranychidae)"J Ethol. 19. 55-62 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawamura N, Yamashiki N, Sahara K: "A cause of male sterility in polyploid silkworms"Dev Growth Differ.. 43(Suppl). S122 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Marec F, Sahara K, Traut W: "Sex chromosomes in Ephestia kuehniella (Insecta, Lepidoptera) tolerate a telomere loss"Ann Genet (Paris). 44(Suppl). S131 (2001)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2004-04-14  

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