Research Unit, Molecular Neuroprotection Group, Hospital Nacional de Parapléjicos, SESCAM, 45071 Toledo, Spain.
Research Unit, Functional Exploration and Neuromodulation of the Central Nervous System (FENNSI) Group, Hospital Nacional de Parapléjicos, SESCAM, 45071 Toledo, Spain.
Int J Mol Sci. 2023 Nov 20;24(22):16509. doi: 10.3390/ijms242216509.
Neuronal maturation is a process that plays a key role in the development and regeneration of the central nervous system. Although embryonic brain development and neurodegeneration have received considerable attention, the events that govern postnatal neuronal maturation are less understood. Among the mechanisms influencing such neuronal maturation processes, apoptosis plays a key role. Several regulators have been described to modulate apoptosis, including post-transcriptional regulation by microRNAs. This study aimed to analyze endogenous expression changes of miR-138-5p, as well as its main validated pro-apoptotic target caspase3, during the maturation of neuronal cultures and their response under apoptotic challenge. Our results point out that the observed opposite expression of miR-138-5p and its target caspase3 might modulate apoptosis favoring neuronal survival at distinct maturation stages. The unchanged expression of miR-138-5p in mature neurons contrasts with the significant downregulation in immature neurons upon apoptotic stimulation. Similarly, immunoblot and individual cellular assays confirmed that during maturation, not only the expression but processing of CASP-3 and caspase activity is reduced after apoptotic stimulation which results in a reduction of neuronal death. Further studies would be needed to determine a more detailed role of miR-138-5p in apoptosis during neuronal maturation and the synergistic action of several microRNAs acting cooperatively on caspase3 or other apoptotic targets.
神经元成熟是中枢神经系统发育和再生过程中的关键步骤。尽管胚胎大脑发育和神经退行性变受到了广泛关注,但调控神经元成熟的过程还知之甚少。在影响这些神经元成熟过程的机制中,细胞凋亡起着关键作用。已经描述了几种调节细胞凋亡的调节剂,包括 microRNAs 的转录后调控。本研究旨在分析神经元培养物成熟过程中内源性 miR-138-5p 的表达变化及其主要的验证性促凋亡靶标 caspase3,并分析其在凋亡挑战下的反应。我们的结果表明,miR-138-5p 和其靶标 caspase3 的观察到的相反表达可能在不同的成熟阶段调节凋亡,有利于神经元存活。成熟神经元中不变的 miR-138-5p 表达与凋亡刺激下未成熟神经元中显著下调形成对比。同样,免疫印迹和单个细胞测定证实,在成熟过程中,凋亡刺激后不仅 CASP-3 的表达而且其加工和 caspase 活性都降低,导致神经元死亡减少。需要进一步研究来确定 miR-138-5p 在神经元成熟过程中细胞凋亡中的更详细作用,以及协同作用于 caspase3 或其他凋亡靶标的几种 microRNAs 的协同作用。