Moisenovich Mikhail M, Silachev Denis N, Moysenovich Anastasia M, Arkhipova Anastasia Yu, Shaitan Konstantin V, Bogush Vladimir G, Debabov Vladimir G, Latanov Alexander V, Pevzner Irina B, Zorova Ljubava D, Babenko Valentina A, Plotnikov Egor Y, Zorov Dmitry B
Department of Biology, Lomonosov Moscow State University, Moscow, Russia.
Laboratory of Mitochondrial Structure and Function, A. N. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, Russia.
Front Cell Dev Biol. 2020 Sep 8;8:823. doi: 10.3389/fcell.2020.00823. eCollection 2020.
The existence of niches of stem cells residence in the ventricular-subventricular zone and the subgranular zone in the adult brain is well-known. These zones are the sites of restoration of brain function after injury. Bioengineered scaffolds introduced in the damaged loci were shown to support neurogenesis to the injury area, thus representing a strategy to treat acute neurodegeneration. In this study, we explored the neuroprotective activity of the recombinant analog of spidroin 1 rS1/9 after its introduction into the ischemia-damaged brain. We used nestin-green fluorescent protein (GFP) transgenic reporter mouse line, in which neural stem/progenitor cells are easily visualized and quantified by the expression of GFP, to determine the alterations in the dentate gyrus (DG) after focal ischemia in the prefrontal cortex. Changes in the proliferation of neural stem/progenitor cells during the first weeks following photothrombosis-induced brain ischemia and effects of spidroin rS1/9 in rat primary neuronal cultures were the subject of the study. The introduction of microparticles of the recombinant protein rS1/9 into the area of ischemic damage to the prefrontal cortex leads to a higher proliferation rate and increased survival of progenitor cells in the DG of the hippocampus which functions as a niche of brain stem cells located at a distance from the injury zone. rS1/9 also increased the levels of a mitochondrial probe in DG cells, which may report on either an increased number of mitochondria and/or of the mitochondrial membrane potential in progenitor cells. Apparently, the stimulation of progenitor cells was caused by formed biologically active products stemming from rS1/9 biodegradation which can also have an effect upon the growth of primary cortical neurons, their adhesion, neurite growth, and the formation of a neuronal network. The high biological activity of rS1/9 suggests it as an excellent material for therapeutic usage aimed at enhancing brain plasticity by interacting with stem cell niches. Substances formed from rS1/9 can also be used to enhance primary neuroprotection resulting in reduced cell death in the injury area.
众所周知，成人大脑的脑室下区和颗粒下区存在干细胞龛。这些区域是脑损伤后功能恢复的部位。已表明，引入受损部位的生物工程支架可支持向损伤区域的神经发生，因此是治疗急性神经退行性变的一种策略。在本研究中，我们探讨了蛛丝蛋白1重组类似物rS1/9引入缺血性损伤脑内后的神经保护活性。我们使用巢蛋白-绿色荧光蛋白（GFP）转基因报告小鼠品系，其中神经干/祖细胞可通过GFP表达轻松可视化和定量，以确定前额叶皮质局灶性缺血后齿状回（DG）的变化。光血栓形成诱导的脑缺血后最初几周神经干/祖细胞增殖的变化以及蛛丝蛋白rS1/9对大鼠原代神经元培养物的影响是本研究的主题。将重组蛋白rS1/9的微粒引入前额叶皮质缺血损伤区域，可导致海马DG区祖细胞的增殖率更高且存活率增加，海马DG区是远离损伤区的脑干细胞龛。rS1/9还增加了DG细胞中线粒体探针的水平，这可能反映祖细胞中线粒体数量增加和/或线粒体膜电位增加。显然，祖细胞的刺激是由rS1/9生物降解形成的生物活性产物引起的，这些产物也可能影响原代皮质神经元的生长、它们的黏附、神经突生长以及神经网络的形成。rS1/9的高生物活性表明它是一种通过与干细胞龛相互作用来增强脑可塑性的治疗用途的优良材料。由rS1/9形成的物质也可用于增强初级神经保护作用，从而减少损伤区域的细胞死亡。
