Biosynthesis of maturation-inducing hormone (1-methyladenine) by starfish ovarian follicle cells and its mechanism in inducing oocyte maturation

1.利用高效液相色谱反相和离子交换柱对卵泡细胞提取物进行分析，发现腺嘌呤相关化合物中含有大量ATP。GSS引起了1-MeA的产生和ATP的消耗。GSS作用下消耗的ATP量与1-MeA产生的ATP量相似。这表明ATP可能是海星卵泡细胞生物合成1-MeA的一个底物。1-甲基腺苷生物合成的调控环amp依赖性蛋白激酶（PKA）存在于斑马鱼卵泡细胞中。环状AMP以剂量依赖的方式激活PKA，使70 kda蛋白磷酸化。对70kDa蛋白作用的进一步研究将有助于进一步了解gss刺激海星卵泡细胞产生1-MeA的规律。合成了1-甲基腺嘌呤及其受体1-取代腺嘌呤与具有电子释放或电子吸收作用的官能团的相互作用，并研究了它们对海星卵母细胞成熟的影响。1- β -垂体甲基腺嘌呤和1-氨基腺嘌呤作为1-MeA的激动剂被新发现诱导卵母细胞成熟。相比之下，1-（对硝基苯）腺嘌呤和1- α -萘基甲基腺嘌呤作为1- mea的拮抗剂抑制了1- mea诱导的卵母细胞成熟。在此基础上，我们尝试合成1-MeA衍生物，用于卵母细胞表面1-MeA受体的亲和标记。

1.Sunstrate for 1-methyladenine biosynthesisAnalyzes using high-performance liquid chromatography of follicel cell extract by reversed-phase and ion-exchange columns indicated a large source of ATP among the adenine-related compounds. GSS caused both 1-MeA production and ATP consumption. The amount of ATP consumed under the influence of GSS was similar to that of 1-MeA produced. These suggest that ATP is a possiblr substrate in the biosynthesis of 1-MeA by starfish ovarian follicle cells.2.Regulation of 1-methyladenine biosynthesisCyclic AMP-dependent protein kinase (PKA) was present in stafish follicle cells. The PKA was activated by cyclic AMP in a dose-dependent manner and 70-kDa protein was phosphorylated. Further studies on the role of the 70kDa protein should provide additional insight into the regular of GSS-stimulated 1-MeA production by starfish follicle cells.3.Interaction between 1-methyladenine and its receptor1-Substituted adenines with functional groups bearing electron-releasing or electron withdrawing effect were synthesized and examined effects on starfish oocyte maturation. 1-beta-Nephtylmethyladenine and 1-aminoadenine as agonists of 1-MeA were newly found to induce oocyte maturation. In contrast, 1- (p-nitrobenzyl) adenine and 1-alpha-naphtylmethyladenine as antagonists of 1-MeA inhibited 1-MeA-induced oocyte maturation. Based on these results, we attempt to synthesize 1-MeA derivatives for affinity label to the 1-MeA receptor on oocyte surface.