Pushchina Evgeniya V, Stukaneva Maria E, Varaksin Anatoly A
Zhirmunsky National Scientific Center of Marine Biology, Far Eastern Branch, Russian Academy of Sciences, 690041 Vladivostok, Russia.
Int J Mol Sci. 2020 Dec 17;21(24):9638. doi: 10.3390/ijms21249638.
Fish are a convenient model for the study of reparative and post-traumatic processes of central nervous system (CNS) recovery, because the formation of new cells in their CNS continues throughout life. After a traumatic injury to the cerebellum of juvenile masu salmon, , the cell composition of the neurogenic zones containing neural stem cells (NSCs)/neural progenitor cells (NPCs) in the acute period (two days post-injury) changes. The presence of neuroepithelial (NE) and radial glial (RG) neuronal precursors located in the dorsal, lateral, and basal zones of the cerebellar body was shown by the immunohistochemical (IHC) labeling of glutamine synthetase (GS). Progenitors of both types are sources of neurons in the cerebellum of juvenile during constitutive growth, thus, playing an important role in CNS homeostasis and neuronal plasticity during ontogenesis. Precursors with the RG phenotype were found in the same regions of the molecular layer as part of heterogeneous constitutive neurogenic niches. The presence of neuroepithelial and radial glia GS+ cells indicates a certain proportion of embryonic and adult progenitors and, obviously, different contributions of these cells to constitutive and reparative neurogenesis in the acute post-traumatic period. Expression of nestin and vimentin was revealed in neuroepithelial cerebellar progenitors of juvenile . Patterns of granular expression of these markers were found in neurogenic niches and adjacent areas, which probably indicates the neurotrophic and proneurogenic effects of vimentin and nestin in constitutive and post-traumatic neurogenesis and a high level of constructive metabolism. No expression of vimentin and nestin was detected in the cerebellar RG of juvenile . Thus, the molecular markers of NSCs/NPCs in the cerebellum of juvenile are as follows: vimentin, nestin, and glutamine synthetase label NE cells in intact animals and in the post-traumatic period, while GS expression is present in the RG of intact animals and decreases in the acute post-traumatic period. A study of distribution of cystathionine β-synthase (CBS) in the cerebellum of intact young showed the expression of the marker mainly in type 1 cells, corresponding to NSCs/NCPs for other molecular markers. In the post-traumatic period, the number of CBS+ cells sharply increased, which indicates the involvement of HS in the post-traumatic response. Induction of CBS in type 3 cells indicates the involvement of HS in the metabolism of extracellular glutamate in the cerebellum, a decrease in the production of reactive oxygen species, and also arrest of the oxidative stress development, a weakening of the toxic effects of glutamate, and a reduction in excitotoxicity. The obtained results allow us to consider HS as a biologically active substance, the numerous known effects of which can be supplemented by participation in the processes of constitutive neurogenesis and neuronal regeneration.
鱼类是研究中枢神经系统(CNS)修复和创伤后恢复过程的便捷模型,因为其CNS中终生都有新细胞形成。在幼年马苏大麻哈鱼的小脑受到创伤性损伤后,急性期(损伤后两天)含有神经干细胞(NSCs)/神经祖细胞(NPCs)的神经源性区域的细胞组成发生变化。通过谷氨酰胺合成酶(GS)的免疫组织化学(IHC)标记显示,位于小脑体背侧、外侧和基底区域的神经上皮(NE)和放射状胶质(RG)神经元前体存在。在幼年个体的小脑组成性生长过程中,这两种类型的祖细胞都是神经元的来源,因此在个体发育过程中的CNS稳态和神经元可塑性中发挥重要作用。具有RG表型的前体在分子层的相同区域中被发现,是异质性组成性神经源性微环境的一部分。神经上皮和放射状胶质GS +细胞的存在表明胚胎和成年祖细胞的一定比例,并且显然这些细胞在创伤后急性期对组成性和修复性神经发生有不同贡献。在幼年个体的小脑神经上皮祖细胞中发现了巢蛋白和波形蛋白的表达。在神经源性微环境和相邻区域发现了这些标志物的颗粒表达模式,这可能表明波形蛋白和巢蛋白在组成性和创伤后神经发生中的神经营养和促神经发生作用以及高水平的建设性代谢。在幼年个体的小脑RG中未检测到波形蛋白和巢蛋白的表达。因此,幼年个体小脑中NSCs / NPCs的分子标志物如下:波形蛋白、巢蛋白和谷氨酰胺合成酶在完整动物和创伤后时期标记NE细胞,而GS表达在完整动物的RG中存在并在创伤后急性期减少。对完整幼年个体小脑中胱硫醚β-合成酶(CBS)分布的研究表明,该标志物主要在1型细胞中表达,与其他分子标志物的NSCs / NCPs相对应。在创伤后时期,CBS +细胞的数量急剧增加,这表明HS参与了创伤后反应。3型细胞中CBS的诱导表明HS参与了小脑中细胞外谷氨酸的代谢、活性氧产生的减少,以及氧化应激发展的停止、谷氨酸毒性作用的减弱和兴奋性毒性的降低。获得的结果使我们能够将HS视为一种生物活性物质,其众多已知作用可通过参与组成性神经发生和神经元再生过程得到补充。