Paul Susana N, De Visser Anna, Motta Federica, Rivers Caroline A, Pooley John R, Lightman Stafford L, Meijer Onno C
Department of Medicine, Division of Endocrinology, Leiden University Medical Center, Leiden, Netherlands; Einthoven Laboratory for Experimental Vascular Medicine, Leiden University Medical Center, Leiden, Netherlands; Translational Health Sciences, Bristol Medical School, University of Bristol, Bristol, United Kingdom.
Department of Medicine, Division of Endocrinology, Leiden University Medical Center, Leiden, Netherlands; Einthoven Laboratory for Experimental Vascular Medicine, Leiden University Medical Center, Leiden, Netherlands.
Steroids. 2025 Feb;214:109524. doi: 10.1016/j.steroids.2024.109524. Epub 2024 Oct 28.
Mineralocorticoid (MR) and glucocorticoid receptors (GR) act as transcription factors and major mediators of glucocorticoid signalling, with pivotal roles in regulating the stress response and hormonal signalling, mood, cognition and memory. The MR and GR share many target genes, have a high degree of homology in their DNA binding (DBD) and ligand binding domain (LBD) but differ considerably in the N-terminal domain (NTD). Using Proximity Ligation Assay (PLA) we quantitatively assessed MR-GR complex subcellular localisation and transcriptional regulation in murine neuroblastoma (N2A) cells stimulated by constant or pulsatile corticosterone (CORT) patterns. We observe that continuous receptor activation by CORT caused localisation at the periphery of the cell nucleus. Truncation of the receptor Ligand Binding Domain (LBD) led to a stronger localisation of MR-GR complexes at the periphery of the cell nuclei. This was also observed for GR immunofluorescence (IF), while in cells expressing only MR or GR the mRNA response to pulsatile hormone treatment was substantially attenuated. However, there was no clearcut correlation between the spatial distribution of MR-GR complexes and the mRNA levels of target genes. Overall, our findings suggest that longer presence in the cell nucleus favors more peripheral nuclear localisation.
盐皮质激素（MR）和糖皮质激素受体（GR）作为转录因子和糖皮质激素信号传导的主要介质，在调节应激反应、激素信号传导、情绪、认知和记忆方面发挥着关键作用。MR和GR共享许多靶基因，在其DNA结合域（DBD）和配体结合域（LBD）中具有高度同源性，但在N端结构域（NTD）中差异很大。我们使用邻近连接分析（PLA）定量评估了在持续或脉冲性皮质酮（CORT）模式刺激下的小鼠神经母细胞瘤（N2A）细胞中MR-GR复合物的亚细胞定位和转录调控。我们观察到，CORT持续激活受体导致其定位于细胞核周边。受体配体结合域（LBD）的截短导致MR-GR复合物在细胞核周边的定位更强。GR免疫荧光（IF）也观察到了这一点，而在仅表达MR或GR的细胞中，对脉冲性激素处理的mRNA反应显著减弱。然而，MR-GR复合物的空间分布与靶基因的mRNA水平之间没有明确的相关性。总体而言，我们的研究结果表明，在细胞核中停留时间越长，越有利于更外周的核定位。
