Characterization of non-olfactory GPCRs in human sperm with a focus on GPR18
Characterization of non-olfactory GPCRs in human sperm with a focus on GPR18
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DOI：
10.1038/srep32255
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发表时间：
2016-08-30
影响因子：
4.6
通讯作者：
Gisselmann, Gunter
中科院分区：
文献类型：
--
作者：
Flegel, Caroline;Vogel, Felix;Gisselmann, Gunter
G蛋白偶联受体（gpcr）将外部化学信号转导为细胞内信号，并参与了大量的生理过程，但关于这些受体在精子中的功能的知识有限。在本研究中，我们进行了RNA-Seq，分析了除嗅觉受体外的所有gpcr在人类精子中的表达。我们在人类精子中发现了多达223种不同的GPCR转录本（FPKM > 0.1），并鉴定出GPR18（一种新发现的大麻素受体）与GPR137和GPR135一起是三种高表达的GPCR。迄今为止，GPR18在人类精子中的表达完全未知。我们通过RT-PCR和免疫细胞化学实验证实了GPR18的表达，并将GPR18蛋白定位在人精子的中间部分。用GPR18配体n -花生四烯酮酰甘氨酸刺激人类精子可诱导12种蛋白激酶的磷酸化，其中一些已知参与顶体反应。与此相一致，n-花生四烯酰基甘氨酸通过改变f -肌动蛋白水平并诱导人精子顶体反应，以浓度依赖性的方式影响细胞骨架。我们的研究结果表明，GPR18可能参与了人类精子的生理过程，提示GPR18可能是精子生理的潜在参与者。
G protein-coupled receptors (GPCRs) transduce external chemical cues into intracellular signals and are involved in a plethora of physiological processes, but knowledge regarding the function of these receptors in spermatozoa is limited. In the present study, we performed RNA-Seq and analyzed the expression of the all GPCRs except olfactory receptors in human spermatozoa. We revealed the expression of up to 223 different GPCR transcripts in human spermatozoa (FPKM > 0.1) and identified GPR18, a newly described cannabinoid receptor, together with GPR137 and GPR135, as one of the three most highly expressed GPCRs. To date, the expression of GPR18 was completely unknown in human spermatozoa. We confirmed GPR18 expression using RT-PCR and immuncytochemistry experiments and localized the GPR18 protein in the midpiece of human spermatozoa. Stimulation of human spermatozoa with the GPR18 ligand N-arachidonoylglycine induced the phosphorylation of 12 protein kinases, some of them are for example known to be involved in the acrosome reaction. In line with this, N-arachidonoylglycine affected the cytoskeleton by changing levels of F-actin and inducing the acrosome reaction in human spermatozoa in a concentration-dependent manner. Our results indicate that GPR18 might be involved in physiological processes of human spermatozoa, suggesting GPR18 to be a potential player in sperm physiology.