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卵子成熟時に獲得する哺乳類初期胚でのアポトーシス回避機構の解析

卵母细胞成熟过程中获得的早期哺乳动物胚胎的凋亡避免机制分析

基本信息

批准号：
12770936
负责人：
鈴木治
金额：
$ 1.79万
依托单位：
National Institute of Infectious Diseases
依托单位国家：
日本
项目类别：
Grant-in-Aid for Encouragement of Young Scientists (A)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

项目摘要

卵子形成時には胚発生能が獲得されるが、卵子成熟培養系の向上にはその獲得機構の解明は非常に重要な課題である。既にBDF1雌マウス17日齢由来卵子と24日齢由来卵子を体外成熟させ、体外受精、体外培養を行なうと胚発生能(具体的には胚盤胞形成能)に有意な差があることを見いだしている。そこで、この発生能の差異を利用して、卵子形成時における胚発生能関連遺伝子の検索を行なった。17および24日齢のBDF1雌マウス16匹ずつより得られた体外成熟卵子(それぞれ、538個と531個)より得たRNAからcDNAライブラリを作成し、遺伝子発現パターンの違いをDifferential display法により調べた。17日齢由来卵子と24日齢由来卵子との間でPCRによる増殖量に差があるPCR産物58個を単離し、サンガー法によってPCR産物のDNA配列を直接決定出来たものは15個であった(現在も配列決定を継続中)。その内、既知の配列は8個で、残りは未報告、もしくは未同定配列(配列は決定されているが遺伝子の性質は未知)であった。卵子で発現することが知られているGDF-9のほか、増殖因子のHDGF、細胞分裂関連遺伝子のEg5等が見られた。ただし、現時点では候補を見いだしたに過ぎず、これらの遺伝子が真の差異発現遺伝子かどうかは、今後ノーザン解析やReal-Time PCR等により、正確に定量する必要がある。前年度報告したようにBDF1においてはB6SJLF1に比べ胚発生時のアポトーシスの関与は現れにくいと思われるが、BDF1卵子においてアポトーシス時に働くcaspaseの基質となりうるdynein light chain遺伝子の差異発現が検出され、卵胞成熟にアポトーシス関連遺伝子の発現が関与する可能性が示された点は非常に興味深いと思われる。

When the egg is formed, the embryo is able to generate energy, and the egg maturation system is able to understand that it is very important. Since the BDF1 female was born on the 17th, the egg on the 24th, the egg on the 24th day, the egg on the 24th, in vitro fertilization, in vitro fertilization, in vitro culture, embryo formation (specific embryo cell formation energy). During the formation of eggs, the embryos were used to generate energy, and the embryos were used to generate energy. Sixteen oocytes matured in vitro were successfully harvested in BDF1 oocytes on the 24th day of gestation, and the in vitro matured oocytes were successfully obtained in RNA cycles, cDNA cycles were performed, and the embryos were tested by Differential display method. On the 17th and 24th, the oocytes were collected from the oocytes on the 24th day. 58 samples were isolated from the oocytes, 58 samples were isolated from the PCR, and the 15 samples were directly determined by the DNA column of the PCR products. (it is now in the queue decision). There are 8 unreported, disabled, and unassigned columns in the internal, known, and unassigned columns. (the column determines that the attribute is unknown). The eggs were divided into four groups: GDF-9, HDGF, mitosis, Eg5, and so on. In the future, we will analyze the Real-Time PCR and other information in the future, and make sure that we can make sure that the necessary information is available. In the previous year's report, there was a significant difference between BDF1 and dynein light chain in the previous year's report. the results showed that there was a difference between pregnancy and pregnancy in the first year of the year, and that in the previous year's report, in the previous year's report, the results of the previous year's report were compared with those in the previous year. The oocyte matured, the possibility, the possibility.