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

1. 1-甲基腺嘌呤生物合成的底物通过反相和离子交换柱对卵泡细胞提取物进行高效液相色谱分析，表明腺嘌呤相关化合物中有大量ATP来源。GSS引起1-MeA的产生和ATP的消耗。在GSS的影响下消耗的ATP的量与产生的1-MeA的量相似。表明ATP可能是海星卵泡细胞合成1-MeA的底物。2. 1-甲基腺嘌呤生物合成的调控海星卵泡细胞中存在环腺苷酸依赖性蛋白激酶（cyclicamp dependent protein kinase，PKA）。PKA被环AMP以剂量依赖性方式激活，70-kDa蛋白被磷酸化。进一步研究70 kDa蛋白的作用，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。3. 1-甲基腺嘌呤与其受体的相互作用合成了具有放电子或吸电子功能的1-取代腺嘌呤，并研究了其对海星卵母细胞成熟的影响。1-β-氨甲酰基腺嘌呤和1-氨基腺嘌呤作为1-MeA的激动剂是新近发现的诱导卵母细胞成熟的药物。相反，1-（对硝基苄基）腺嘌呤和1-α-萘甲基腺嘌呤作为1-MeA的拮抗剂抑制1-MeA诱导的卵母细胞成熟。基于这些结果，我们尝试合成1-MeA衍生物，用于亲和标记卵母细胞表面的1-MeA受体。

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.