Molecular mechanism of evolution and morphological diversity in Cladocera.

枝角类进化和形态多样性的分子机制。

• 批准号: 09839034
• 负责人: YAMAGATA Hideo
• 金额: $ 1.73万
• 依托单位: Tokyo University of Pharmacy and Life Science
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09839034/
• 关键词: Dephnia; Hemoglobin; Hypoxia response; Morphogenic genes; Embryo development; 分子進化; 形態形成; ホメオボックス; 遺伝子解析; 形態多様性

(1) Hemoglobin (Hb) purified from the water flea, Daphnia magna, was analyzed by two-dimensional gel electrophoresis. The Hb was shown to be composed of 6 major subunit chain species. The NHィイD22ィエD2-terminal amino acid sequences of the 6 Hb chains showed that at least 4 Hb genes are present in D. magna. The Hb genes were differentially regulated by the ambient oxygen concentration. Four Hb genes constituted a cluster on the chromosome of D. magna. The complete nucleotide sequences of these genes and their cDNAs showed that the genes have a seven-exon, six-intron structre. A cDNA homologous to that for Arnt, a subunit of transcriptional factor HIF1α which plays a major role in response to hypoxia in mammalian cells, was isolated from D. magna and then its complete nucleotide sequence was determined.(2) cDNAs homologous to those of Drosophila Hox genes, Antp, Ubx, Abd-A, and Scr, and other developmental genes, dll, wnt-1, and hedgehog, were isolated from D. magna and then their complete uncleotide sequences were determined. Expression patterns of these cDNAs during development of D. magna embryos were analyzed by whole mount in situ hybridization. Daphnia Antp cDNA was expressed in Drosophila embryos using the Gal4/UAS system. While it retained the same functions as those of Drosophila Antp, such as the ectopical induction of teashirt and decapentaplegic genes, it showed some functions different from those of Drosophila Antp.

(1)采用双向凝胶电泳技术对从大型蚤（Daphnia magna）中纯化的血红蛋白（Hb）进行了分析。血红蛋白由6种主要亚基链组成。6条血红蛋白链的NH → D22 → D2端氨基酸序列分析表明，D.麦格纳Hb基因的差异调节的环境氧浓度。4个Hb基因在D.麦格纳这些基因的核苷酸序列和cDNA序列分析表明，它们具有7个外显子、6个内含子的结构。从D. magna，然后测定其完整的核苷酸序列。(2)从果蝇中分离到与果蝇Hox基因Antp、Ubx、Abd-A和Scr以及其它发育基因dll、wnt-1和hedgehog同源的cDNA。magna，并测定其核苷酸全序列。这些cDNA在D.用全胚原位杂交技术分析麦格纳胚。利用Gal 4/UAS系统在果蝇胚胎中表达水蚤Antp cDNA。虽然它保留了与果蝇Antp相同的功能，如对teashirt和decapentaplegic基因的异位诱导，但它显示出与果蝇Antp不同的功能。

项目成果

Shoko Kimura: "Heterogeneity and differential expression under hypoxia of two-domain hemoglobin chains in the water flea,Daphnia magna"Journal of Biological Chemistry. 274・15. 10649-10653 (1999)

Shoko Kimura：“水蚤中两域血红蛋白链的异质性和差异表达，大型水蚤”生物化学杂志 274・15（1999）。

Toshihiro Ohta: "An assay system detecting environmental toxicants with cultured Cladoceran eggs in vitro: Malformations induced by ethylene-thiourea." Environmental Research. 77. 43-48 (1998)

Toshihiro Ohta：“用体外培养的枝角类卵检测环境毒物的检测系统：乙烯硫脲引起的畸形。”

Kobayashi, M., and Yamagata, H.: "Structure and function of Hemoglobin in Daphnia magna."Trends in Comparative Biochemistry & Physiology. (in press).

Kobayashi, M. 和 Yamagata, H.：“大型水蚤中血红蛋白的结构和功能。”比较生物化学的趋势

山形 秀夫: "愛嬌ある容姿と抜群の環境適応能力 : ミジンコ." 実験医学. 16. 927-927 (1998)

山形秀夫：“迷人的外表和出色的环境适应能力：实验医学。”16. 927-927 (1998)

志賀 靖弘: "ミジンコの可能性" 生命誌. 6・2. 20-21 (1998)

志贺泰弘：“水蚤的可能性”生物学杂志 6・2（1998）。