Soga Tomoko, Lim Wei Ling, Khoo Alan Soo-Beng, Parhar Ishwar S
Brain Research Institute, School of Medicine and Health Sciences, Monash University Malaysia , Bandar Sunway , Malaysia.
Molecular Pathology Unit, Cancer Research Centre, Institute for Medical Research , Kuala Lumpur , Malaysia.
Front Endocrinol (Lausanne). 2016 Mar 1;7:15. doi: 10.3389/fendo.2016.00015. eCollection 2016.
Kisspeptin, a newly discovered neuropeptide, regulates gonadotropin-releasing hormone (GnRH). Kisspeptins are a large RF-amide family of peptides. The kisspeptin coded by KiSS-1 gene is a 145-amino acid protein that is cleaved to C-terminal peptide kisspeptin-10. G-protein-coupled receptor 54 (GPR54) has been identified as a kisspeptin receptor, and it is expressed in GnRH neurons and in a variety of cancer cells. In this study, enhanced green fluorescent protein (EGFP) labeled GnRH cells with migratory properties, which express GPR54, served as a model to study the effects of kisspeptin on cell migration. We monitored EGFP-GnRH neuronal migration in brain slide culture of embryonic day 14 transgenic rat by live cell imaging system and studied the effects of kisspeptin-10 (1 nM) treatment for 36 h on GnRH migration. Furthermore, to determine kisspeptin-induced molecular pathways related with apoptosis and cytoskeletal changes during neuronal migration, we studied the expression levels of candidate genes in laser-captured EGFP-GnRH neurons by real-time PCR. We found that there was no change in the expression level of genes related to cell proliferation and apoptosis. The expression of ankyrin repeat domain-containing protein (ankrd) 26 in EGFP-GnRH neurons was upregulated by the exposure to kisspeptin. These studies suggest that ankrd 26 gene plays an unidentified role in regulating neuronal movement mediated by kisspeptin-GPR54 signaling, which could be a potential pathway to suppress cell migration.
kisspeptin是一种新发现的神经肽,可调节促性腺激素释放激素(GnRH)。Kisspeptins是一个大型的RF-酰胺肽家族。由KiSS-1基因编码的kisspeptin是一种145个氨基酸的蛋白质,可被切割成C末端肽kisspeptin-10。G蛋白偶联受体54(GPR54)已被鉴定为kisspeptin受体,它在GnRH神经元和多种癌细胞中表达。在本研究中,用增强型绿色荧光蛋白(EGFP)标记具有迁移特性且表达GPR54的GnRH细胞,作为研究kisspeptin对细胞迁移影响的模型。我们通过活细胞成像系统监测了胚胎第14天转基因大鼠脑片培养中EGFP-GnRH神经元的迁移,并研究了kisspeptin-10(1 nM)处理36小时对GnRH迁移的影响。此外,为了确定kisspeptin诱导的与神经元迁移过程中细胞凋亡和细胞骨架变化相关的分子途径,我们通过实时PCR研究了激光捕获的EGFP-GnRH神经元中候选基因的表达水平。我们发现与细胞增殖和凋亡相关的基因表达水平没有变化。暴露于kisspeptin后,EGFP-GnRH神经元中含锚蛋白重复结构域蛋白(ankrd)26的表达上调。这些研究表明,ankrd 26基因在调节由kisspeptin-GPR54信号介导的神经元运动中发挥未知作用,这可能是抑制细胞迁移的潜在途径。