Department of Experimental Medicine, University of Campania "Luigi Vanvitelli", 80138 Naples, Italy.
Department of Environmental, Biological, Pharmaceutical Sciences and Technologies, University of Campania "Luigi Vanvitelli", 81100 Caserta, Italy.
Int J Biol Sci. 2022 Aug 8;18(13):5136-5153. doi: 10.7150/ijbs.76261. eCollection 2022.
CircRNA cargo in spermatozoa (SPZ) participates in setting cell quality, in terms of morphology and motility. Cannabinoid receptor CB1 activity is correlated with a proper spermatogenesis and epididymal sperm maturation. Despite CB1 promotes endogenous skill to circularize mRNAs in SPZ, few notions are reported regarding the functional link between endocannabinoids and spermatic circRNA cargo. In CB1 knock-out male mice, we performed a complete dataset of spermatic circRNA content by microarray strategy. Differentially expressed (DE)-circRNAs, as a function of genotype, were identified. Within DE-circRNAs, we focused the attention on circLIMA1, as putative actin-cytoskeleton architecture regulator. The validation of circLIMA1 dependent-competitive endogenous RNA (ceRNA) network (ceRNET) in cell line confirmed its activity in the regulation of the cytoskeletal actin. Interestingly, a dynamic actin regulation in SPZ nuclei was found during their epididymal maturation. In this scenario, we showed for the first time an intriguing sperm nuclear actin remodeling, regulated a ceRNET-independent pathway, consisting in the nuclear shuttling of circLIMA1-QKI interactome and downstream in Gelsolin regulation. In particular, the increased levels of circLIMA1 in CB1 knock-out SPZ, associated with an inefficient depolymerization of nuclear actin, specifically illustrate how endocannabinoids, by regulating circRNA cargo, may contribute to sperm morpho-cellular maturation.
精子(SPZ)中的环状 RNA 货物参与设定细胞质量,包括形态和运动性。大麻素受体 CB1 的活性与适当的精子发生和附睾精子成熟有关。尽管 CB1 促进 SPZ 中内源性 mRNA 环化,但关于内源性大麻素和精子环状 RNA 货物之间的功能联系的报道很少。在 CB1 敲除雄性小鼠中,我们通过微阵列策略对精子环状 RNA 含量进行了全面数据集分析。鉴定了基因型相关的差异表达(DE)-环状 RNA。在 DE-环状 RNA 中,我们将注意力集中在环状 LIMA1 上,因为它是一种潜在的肌动蛋白细胞骨架结构调节剂。在细胞系中对环状 LIMA1 依赖性竞争性内源 RNA(ceRNA)网络(ceRNET)的验证证实了其在调节细胞骨架肌动蛋白中的活性。有趣的是,在它们的附睾成熟过程中,在 SPZ 核中发现了一种动态的肌动蛋白调节。在这种情况下,我们首次展示了一种有趣的精子核肌动蛋白重塑,受 ceRNET 独立途径调节,该途径由环状 LIMA1-QKI 相互作用体的核穿梭和下游的 Gelsolin 调节组成。特别是,在 CB1 敲除 SPZ 中,circLIMA1 水平增加与核肌动蛋白解聚效率降低有关,这特别说明了内源性大麻素如何通过调节环状 RNA 货物来促进精子形态细胞成熟。