Molecular mechanisms of oviposition control

产卵控制的分子机制

基本信息

批准号：
04044079
负责人：
SHIMADA Kiyoshi
金额：
$ 5.89万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1993
项目状态：
已结题

项目摘要

We have proposed a hypothesis for oviposition mechanism as follows ; ovarian prostaglandin is released into peripheral plasma to stimulating uterine contractions, which in turn, triggers AVT release from the neurohypophysis. The present project was conducted to prove the above hypothesis by providing following evidence on the molecular basis.1. Prostaglandins were produced in the theca layers of the largest pre- and postovulatory follicles and tissue PG level was highest with concurrent increase in plasma PGF concentration immediately before oviposition. Ovarian-oviposition inducing factor was identified as PG by HPLC analysis. These results indicate that ovarian PG plays a pivotal role in the regulation of oviposition.2. Plasma AVT concentration also increased at the time of oviposition. AVT was released in response to uterine contractions but this increase was suppressed by indomethacin treatment which delayed oviposition. Immunocytochemical studies revealed that AVT-containing neuro … More ns were detected at the preoptic region and superficial area of the third ventricle. AVT cDNA was cloned and its base sequence was determined. Using the 562-base cDNA as a probe Northern hybridization analysis detected about 700 bases of AVT mRNA in the hypothalamus. The mRNA levels increased 4-fold after 4-day of water deprivation. Analysis of in situ hybridization revealed AVT mRNA localized in the periventricular nuclei of the hypothalamus.3. Finally, the project of cloning of AVT receptor cDNA is stil in progress but solubilized fraction of AVT receptor of the chicken uterus was purified, electophoresed and this fraction was isolated. At present, amino acid sequencing is under an investigation and will be used for primers in PCR cloning to isolate the gene. On the basis of amino acid sequences of homologous region of human oxytocin receptor cDNA and rat arginine vasoporessin receptor, primers were synthesized for RT-PCR of chicken uterus RNA. The 452-bases RT-PCR cDNA product was used as a probe for Northern hybridization of the chicken uterus. Hybridizable mRNA species of 5.6, 4.6 and 2.1 kd were identified as AVT receptor mRNA. Less

我们提出了一个关于产卵机制的假设，如下所示。卵巢前列腺素被释放到外周血浆中，以刺激子宫收缩，进而触发了AVT从神经hohypophysy中释放出来。本项目是通过根据分子提供以下证据的上述假设1。前列腺素是在卵巢前和卵巢前和组织后最大的theca层中产生的，而组织PG水平最高，同时同时增加血浆PGF浓度，直到产卵前。通过HPLC分析将卵巢卵巢诱导的因子鉴定为PG。这些结果表明，卵巢PG在产卵的调节中起关键作用。2。血浆AVT浓度在排卵时也会增加。 AVT是为了响应子宫收缩而释放的，但由于抑制产卵的吲哚美辛治疗而抑制了这种增加。免疫细胞化学研究表明，在第三次通气的前区域和浅表区域中检测到含AVT的神经……更多的NS。克隆AVT cDNA，并确定其碱基序列。使用562碱cDNA作为探针北部杂交分析，检测到下丘脑中约有700个AVT mRNA。缺水4天后，mRNA水平升高了4倍。对原位杂交的分析表明，AVT mRNA位于下丘脑的周期核中。3。最后，AVT受体cDNA克隆的项目正在进行中，但是纯化了鸡子宫的AVT受体的可溶性部分，纯化了，电载体并分离了该部分。目前，氨基酸测序正在研究中，将用于PCR克隆中的底漆以分离基因。根据人氧受体cDNA的同源区域和大鼠精氨酸血管雷素受体的氨基酸序列，将底漆用于鸡子宫RNA的RT-PCR。 452-Bases RT-PCR cDNA产物用作鸡子宫北部杂交的探针。可鉴定为5.6、4.6和2.1 kd的杂交mRNA物种为AVT受体mRNA。较少的