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Acetylcholine/acetylcholine esterase and necroptosis in the ovary

乙酰胆碱/乙酰胆碱酯酶和卵巢坏死性凋亡

基本信息

批准号：
195528179
负责人：
Professor Dr. Artur Mayerhofer
金额：
--
依托单位：
Pathologisches Institut
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2011
资助国家：
德国
起止时间：
2010-12-31 至 2019-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/195528179?language=en
关键词：
Acetylcholine acetylcholine esterase necroptosis ovary

项目摘要

Acetylcholine (ACh) is produced in the ovary and exerts trophic actions, which are mediated by receptors. Recent studies on the cholinergic system of the ovary revealed that ACh-esterase (AChE) is also produced with the ovary, specifically by granulosa cell (GCs) and that it counteracts the trophic actions of ACh. AChE can be blocked by pharmacological intervention in cultured human GCs. Furthermore, programmed cell death (necroptosis) was identified in cultured human GCs as an unknown form of cell death, and was linked to a non-enzymatic action of an AChE splice variant, AChE-R. This variant and clear evidence for necroptotic cell death in follicles and the corpus luteum (CL) were found in vivo, in the human and non-human primate ovary. An AChE-R peptide strongly enhanced necroptosis of human GCs, and on the other side, necroptotic cell death of GCs is amenable to pharmacological blockage by necrostatin-1 and necrosulfonamide. Thus, both ACh-E-blockers and necroptosis-blockers may be novel tools to interfere with important ovarian processes (follicular growth, follicular atresia and/or luteolysis). First studies in rats, in which an AChE blocker was applied locally to the ovary, are in full support for a role of ACh in the promotion of follicular growth. It is possible that besides normal ovarian events, ovarian pathologies, or the outcome of follicular cultures could be targeted. We now propose a series of studies to examine these possibilities. We will analyze sites of necrotic/necroptotic events and their frequency in human and non-human primate ovary by immunohistochemistry, using a novel marker (p-MLKL). Apoptosis markers will allow us to distinguish between necroptosis and apoptosis. Since AChE-R can induce necroptosis in GCs, we will study, whether AChE-R expression correlates with p-MLKL in ovarian sections. Studies in human GC-tumors will be performed as well. To explore regulation and actions of AChE-R, we will use human IVF-derived GCs. We will among others examine a role of miRNA132 and its hormonal regulation and will attempt to elucidate the mechanisms of action of ARP-induced necroptosis. Finally, through international collaboration we will examine, whether pharmacological intervention with necroptosis improves the survival of cultured monkey follicles and may influence follicular fate in rat. We expect important translational insights into an unexplored form of cell death and its role in ovarian physiology and pathology, insighty which may open novel doors to interfere with ovarian events.

乙酰胆碱（ACh）在卵巢中产生，并通过受体介导发挥营养作用。最近对卵巢胆碱能系统的研究表明，乙酰胆碱酯酶（AChE）也与卵巢，特别是颗粒细胞（GCs），它抵消了乙酰胆碱的营养作用。在培养的人GC中，AChE可通过药物干预被阻断。此外，程序性细胞死亡（坏死性凋亡）在培养的人GC中被鉴定为未知形式的细胞死亡，并与AChE剪接变体AChE-R的非酶促作用有关。在人类和非人类灵长类动物卵巢中，在体内发现了卵泡和黄体（CL）中坏死性细胞死亡的这种变体和明确证据。 AChE-R肽强烈增强人GC的坏死性凋亡，另一方面，GC的坏死性凋亡细胞死亡可通过necrostatin-1和necrosulfonamide进行药理学阻断。因此，乙酰胆碱酯酶阻断剂和坏死性凋亡阻断剂都可能是干扰重要卵巢过程（卵泡生长、卵泡闭锁和/或黄体溶解）的新工具。在大鼠中进行的第一项研究中，将AChE阻滞剂局部应用于卵巢，完全支持ACh在促进卵泡生长中的作用。除了正常的卵巢事件外，卵巢病理或卵泡培养的结果也可能成为目标。我们现在提出一系列研究来检验这些可能性。我们将使用一种新的标记物（p-MLKL），通过免疫组化分析人类和非人类灵长类动物卵巢中坏死/坏死性凋亡事件的发生部位及其频率。凋亡标志物将使我们能够区分坏死性凋亡和凋亡。由于AChE-R可诱导GC的坏死性凋亡，我们将研究AChE-R表达是否与卵巢切片中的p-MLKL相关。还将进行人GC肿瘤研究。为了探索AChE-R的调节和作用，我们将使用人IVF衍生的GC。我们将研究miRNA 132及其激素调节的作用，并试图阐明ARP诱导的坏死性凋亡的作用机制。最后，通过国际合作，我们将研究药物干预坏死性凋亡是否能提高培养猴卵泡的存活率，并可能影响大鼠卵泡的命运。我们期待着对一种未探索的细胞死亡形式及其在卵巢生理学和病理学中的作用的重要翻译见解，这可能会为干扰卵巢事件打开新的大门。