Lupacchini Leonardo, Mollinari Cristiana, Tancredi Virginia, Garaci Enrico, Merlo Daniela
IRCCS, San Raffaele Roma, Via di Val Cannuta.
Institute of Translational Pharmacology (IFT), National Research Council.
Neuroreport. 2025 Apr 2;36(6):290-296. doi: 10.1097/WNR.0000000000002149. Epub 2025 Mar 26.
DNA-dependent protein kinase catalytic subunit (DNA-PKcs) is one of the key enzymes involved in DNA double-strand break (DSB) repair. However, recent studies using DNA-PKcs knockout mice revealed that DNA-PKcs plays an important role in neuronal plasticity. The aim of this study was to examine the role of DNA-PKcs on synaptic plasticity in severe combined immunodeficiency disease (SCID) mice that carry a mutation resulting in a DNA-PKcs protein devoid of kinase activity but still expressed in cells, although with a small COOH-terminal truncation. To this aim, we carried out electrophysiological and molecular analysis on hippocampal slices from wild-type (WT) and SCID mice. Electrophysiological analysis showed an impairment in the basal synaptic transmission in SCID mice compared with WT, whereas paired-pulse facilitation, caused by presynaptic mechanisms, was not different in the two groups of animals. By contrast, tetanic stimulation induced long-term potentiation (LTP) with values that were approximately 43% lower in slices from SCID mice compared with WT. The same slices used for electrophysiology were analyzed to study the phosphorylation state of cAMP response element-binding protein (CREB) and extracellular signal-regulated kinases and to evaluate mRNA expression levels of CREB-target genes at different times after LTP induction. In conclusion, molecular analysis did not show significant differences between SCID and WT brain slices, thus confirming the evidence that DNA-PKcs kinase activity directly regulates neuronal functions and plays a novel role beyond DSB repair. Moreover, these results indicate that studies using SCID mice involving analysis of synaptic function need to be interpreted with caution.
DNA依赖性蛋白激酶催化亚基(DNA-PKcs)是参与DNA双链断裂(DSB)修复的关键酶之一。然而,最近使用DNA-PKcs基因敲除小鼠的研究表明,DNA-PKcs在神经元可塑性中起重要作用。本研究的目的是检测DNA-PKcs在严重联合免疫缺陷病(SCID)小鼠突触可塑性中的作用,这些小鼠携带一种突变,导致DNA-PKcs蛋白缺乏激酶活性,但仍在细胞中表达,尽管其COOH末端有小的截短。为此,我们对野生型(WT)和SCID小鼠的海马切片进行了电生理和分子分析。电生理分析显示,与WT小鼠相比,SCID小鼠的基础突触传递受损,而由突触前机制引起的双脉冲易化在两组动物中没有差异。相比之下,强直刺激诱导的长时程增强(LTP)在SCID小鼠切片中的值比WT小鼠低约43%。对用于电生理的相同切片进行分析,以研究环磷酸腺苷反应元件结合蛋白(CREB)和细胞外信号调节激酶的磷酸化状态,并评估LTP诱导后不同时间CREB靶基因的mRNA表达水平。总之,分子分析未显示SCID和WT脑切片之间存在显著差异,从而证实了DNA-PKcs激酶活性直接调节神经元功能并在DSB修复之外发挥新作用的证据。此外,这些结果表明,使用SCID小鼠进行涉及突触功能分析的研究需要谨慎解释。