Development of techniques for in vitro production of good quality eggs by elucidation of mechanisms of oocyte cytoplasmic maturation in teleost

通过阐明硬骨鱼卵母细胞胞质成熟机制，开发体外生产优质卵子的技术

基本信息

批准号：
18580187
负责人：
KAGAWA Hirohiko
金额：
$ 2.49万
依托单位：
University of Miyazaki
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

To elucidate mechanisms of oocyte cytoplasmic maturation (such as a hydration, liquid coalescence and clearing of the ooplasm) during oocyte maturation and to develop techniques for in vitro production of good quality eggs, the present study examined the following studies in the Japanese eel spawn buoyant eggs.1. The present study shows the process of oocyte cytoplasmic maturation during de novo oocyte maturation in the Japanese eel. Moreover, we also developed the incubation techniques for mimic the cytoplasmic maturation occurred in vivo. Using the techniques we developed, we found that cytoplasmic maturation was induced by the maturation-inducing steroid that was synthesized in the ovarian follicles by the stimulation of gonadotropin. Cytoplasmic maturation induced by maturation-inducing steroid may be mediated by protein synthesis in the oocyte. IGF-I, but not GAP junction may be involved in the cytoplasmic maturation of eel oocytes.2. The present study also indicates that oocyte h … More ydration is mediated by yolk proteolysis which creates the osmotic driving force and aquaporin possibly facilitates water uptake into oocytes. Moreover, the study indicates that the ovarian follicles in the Japanese eel may play an essential role in oocyte hydration, although the precise role must be identified in the further research.3. The present study also examined the Japanese eel for the presence and cellular distribution of eel AQP-1o in the ovarian follicles during oocyte growth, oocyte maturation and ovulation by the immunocytochemistry. Anti-AQP-1o antibody was raised in a rabbit against a synthetic peptide corresponding to a part of the C-terminal region of eel AQP-1o. Immunoreactivity was first observed in the cytoplasm around the yolk globules which were located in the peripheral region of the oocytes during the primary yolk globule stage; thereafter, the immunoreactivity was observed throughout the cytoplasm of the oocyte as vitellogenesis progressed. The immunoreactivity became localized around the large membrane-limited yolk masses which were formed by the fusion of yolk globules. It also localized below the oocyte plasma membrane in the migratory nucleus stage oocytes, mature oocytes and ovulated eggs. These results are different from those observed in the oocytes of gilthead sea bream and suggest the existence of a distinct and characteristic mechanism of oocyte hydration in the eel.4. We tried to develop the techniques for in vitro production of good quality eggs in the Japanese eel. Although we found the oocyte maturation and ovulation occurred at 12 and 18 hours after incubation with maturation-inducing steroid respectively, we could not obtained fertilized eggs. Further studies are necessary to develop the techniques for obtaining good quality eggs in vitro. Less

为了阐明在卵母细胞成熟过程中卵母细胞细胞质成熟的机制（例如水合，液体结合和卵质的清除），并开发了用于体外生产高质量鸡蛋的技术，本研究检查了日本eEL产生的生物产生卵子的以下研究。本研究显示了日本鳗鱼中从头卵母细胞成熟期间卵母细胞胞质成熟的过程。此外，我们还开发了模仿细胞质成熟的孵育技术。使用我们开发的技术，我们发现细胞质成熟是由成熟诱导的类固醇诱导的，该类固醇是通过促促性腺激素刺激在卵巢诱导中合成的。成熟诱导的类固醇诱导的细胞质成熟可能是由卵母细胞中的蛋白质合成介导的。 IGF-I（而不是间隙连接）可能参与鳗鱼卵母细胞的细胞质成熟。2。本研究还表明，卵母细胞h…更多的y蛋白蛋白水解介导，从而产生渗透驱动力和水通道蛋白可能会促进水吸入卵母细胞。此外，该研究表明，日本鳗鱼中的卵巢手续可能在卵母细胞水合中起着至关重要的作用，尽管必须在进一步的研究中确定精确作用。3。本研究还检查了日本鳗鱼在卵母细胞生长，卵母细胞成熟和通过免疫细胞化学的排卵期间在卵巢福利中的存在和细胞分布。在兔子中升高了抗AQP-1O抗体，针对与EEL AQP-1O的C末端区域相对应的合成肽。首先在原发性蛋黄球期间位于卵母细胞周围的蛋黄球周围的细胞质中观察到免疫反应性。此后，随着植物发生的发展，在整个卵母细胞的整个细胞质中都观察到免疫反应性。免疫反应性在大型膜覆有蛋黄的肿块周围定位，这些蛋黄是由蛋黄球的融合形成的。它也位于卵母细胞质膜下方，迁移核细胞，成熟的卵母细胞和排卵卵。这些结果与在吉尔特黑德海花的卵母细胞中观察到的结果不同，并表明存在卵母细胞水合的独特而特征的机理。4。我们试图开发日本鳗鱼体外生产优质鸡蛋的技术。尽管我们发现卵母细胞成熟和排卵分别在与成熟诱导的类固醇孵育后的12和18小时发生，但我们无法获得受精卵。需要进一步的研究来开发体外获得高质量鸡蛋的技术。较少的

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

オスモティックポンプを用いたサケ脳下垂体抽出液投与が雌ウナギの成熟に及ぼす影響

使用渗透泵施用鲑鱼垂体提取物对雌性鳗鱼成熟的影响

DOI：
发表时间：
2007
期刊：
影响因子：
0
作者：
Fujie;N.;Adachi;J.;Nishi;A.;Hashimoto;H.;Kaji;S.;Kishi;T.;Kagawa;H.;堀内泰宏;香川浩彦
通讯作者：
香川浩彦

Molecular characterization of three forms of vitellogenin and their yolk protein products during oocyte growth and maturation in red seabream (Pagrus majar). a marine teleost spawning pelagic eggs

红鲷（Pagrus majar）卵母细胞生长和成熟过程中三种形式卵黄蛋白原及其卵黄蛋白产物的分子特征。

DOI：
发表时间：
2006
期刊：
Molecular Reproduction and Development 73
影响因子：
0
作者：
Sawaguchi;S.;Kagawa;H.;Ohkubo;N.;Hiramatsu;N.;Sullivan;C.V.;Matsubara;T.;香川浩彦;Ozaki Y.;Sawaguchi S.
通讯作者：
Sawaguchi S.

Expression of growth hormone family and growth hormone receptor during early development in the Japanese eel (Anguilla japonica)

日本鳗鱼（Anguilla japonica）早期发育过程中生长激素家族和生长激素受体的表达