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顕微授精法を用いた、昆虫における精細胞の受精能および人工生殖に関する研究

利用显微授精方法研究昆虫精子细胞的受精能力和人工繁殖

基本信息

批准号：
09780684
负责人：
畠山正統
金额：
$ 1.34万
依托单位：
Kobe University
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：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-09780684/
关键词：
顕微授精 ICSI 受精精細胞キメラ膜翅目昆虫

项目摘要

1カブラハバチ(Athalia rosae、広腰亜目)は昆虫で唯一、顕微授精法(ICSI 法)により、体外人工授精が行える実験系である。この特長を利用して、精巣中の精子形成過程のさまざまなステージにある生殖細胞(精細胞)を成熟未受精卵に注入し、これらの生殖細胞の授精能と発生参加能について調べた。円形精細胞および尾部伸長中の精細胞(elongating spermatid)を注入した場合、これらの精細胞は卵核とは融合せず、単独で独立に発生に参加して、半数体雄キメラ個体が生じた。マーカー突然変異を用いて子孫検定を行うことにより、これらのキメラ雄個体の生殖細胞は卵核由来、精細胞核由来の両方からなっていることがわかった。一方、精巣内の未成熟精子(elonsated spermatid)を用いると受精個体が得られたが、半数体雄キメラ個体は得られなかった。これらの結果から、昆虫(カブラハバチ)の精細胞には発生参加能があり、卵核と独立に発生に参加するか、受精するかは、精子形成過程の段階に依存していることが示された。2精子あるいは精細胞の発生参加に関与する卵細胞質内因子の昆虫間での共通性を調べるために、近縁異種の精細胞を注入した。用いた異種の成熟精子は顕微受精によって胚発生初期まで生存できる雑種を形成する。異種の精細胞を注入すると種間キメラ個体が得られ、雑種個体は得られなかった。これらのキメラ個体は胚発生は完了するが析化には至らなかった。3カブラハバチの未成熟卵巣を近縁異種に移植し、別種の卵黄蛋白質のみで成熟した卵を得て、これらに凍結保存精子を顕微注入した。その結果、通常の顕微授精の場合と同様に受精個体が得られ、昆虫における系統保存に応用できる人工生殖の可能性が示された。

The main results are as follows: (1) the insect is the only one in the order of Athalia rosae, microinsemination (ICSI) and artificial insemination in vitro. In the process of spermatogenesis, the mature unfertilized eggs (spermatocytes) are injected into the mature unfertilized eggs, and the fertilisation of sperm cells can participate in the process of spermatogenesis. The sperm cells (elongating spermatid) in the tail elongation of the spermatozoa were injected into the sperm cells, the egg nuclei of the sperm cells were fused, the sperm cells were born independently, and half of the male embryos were born. All of a sudden, you have to use the baby to determine the origin of the egg nucleus and the sperm nucleus of the male reproductive cell of the mother, the mother and the baby. Immature spermatozoa (elonsated spermatid) of one side and semen were fertilized with semen, and half of the males were fertilized with sperm. The results showed that the sperm cells of insects were able to participate in the process of fertilization, the sperm cells of insects participated in the process of fertilization, the sperm cells of insects participated in the process of spermatogenesis, and the sperm formation in the process of spermatogenesis was detected in the process of sperm formation. (2) spermatozoa were injected into sperm cells to participate in the commonality between spermatozoa and oocyte intracellular factors in insects, and recent sperm cell injection. Several kinds of mature spermatozoa were used to microfertilize the embryos in the early stage of embryo development. Several kinds of fine cells are injected into each other, and several kinds of cells are successfully injected into each other. After the birth of the embryo, we have finished the analysis of the mother to the mother. (3) the immature eggs were harvested in recent years, the mature eggs were harvested in other kinds of yolk protein culture, and the spermatozoa were preserved by microinjection. The results showed that the possibility of artificial reproduction was demonstrated by the possibility of microinsemination and co-fertilization in general, and the possibility of artificial reproduction in insect breeding system.