Bender Melanie, Schwind Lisa, Grundmann David, Martin Monika, Klotz Markus, Götz Claudia, Montenarh Mathias, Schäfer Karl-Herbert
Working Group Enteric Nervous System, University of Applied Sciences Kaiserslautern, Campus Zweibrücken, Germany.
Medical Biochemistry and Molecular Biology, Building 44, University of Saarland, 66421 Homburg Saar, Germany.
Heliyon. 2017 Jun 9;3(6):e00318. doi: 10.1016/j.heliyon.2017.e00318. eCollection 2017 Jun.
Protein kinases play central roles in cell and tissue development. Protein kinase CK2, an ubiquitously expressed serine/threonine kinase has severe impacts on embryo- and spermatogenesis. Since its role in neurogenesis has so far only been investigated in very few studies, we analysed the role of CK2 in neural stem cells by using two specific inhibitors.
Neural stem cells were isolated from the subventricular zone of neonatal mice, using a neurosphere approach. Proliferation of the neurospheres, as well as their differentiation was investigated with and without inhibition of CK2. Changes in proliferation were assessed by counting the number and measuring the diameter of the neurospheres. Furthermore, the absolute cell numbers within the neurospheres were estimated. Differentiation was induced by retinoic acid in single cells after dissociation of the neurospheres. CK2 was inhibited at consecutive time points after induction of the differentiation process.
CK2 inhibition reduced the amount and size of proliferating neurospheres dose dependently. Adding the CK2 inhibitor CX-4945 at the start of differentiation we observed a dose-dependent effect of CX-4945 on cell viability and glia cell differentiation. Adding quinalizarin, a second CK2 inhibitor, at the start of differentiation led to an elevated level of apoptosis, which was accompanied by a reduced neural differentiation. Adding the CK2 inhibitors at 72 h after the start of differentiation had no effect on stem cell differentiation. Conclusion: Inhibition of CK2 influences early gliogenesis in a time point and concentration dependent manner.
The use of a CK2 inhibitor significantly affects the neural stem cell niche.
蛋白激酶在细胞和组织发育中起核心作用。蛋白激酶CK2是一种广泛表达的丝氨酸/苏氨酸激酶，对胚胎发育和精子发生有严重影响。由于其在神经发生中的作用迄今仅在极少数研究中得到调查，我们使用两种特异性抑制剂分析了CK2在神经干细胞中的作用。
采用神经球法从新生小鼠的脑室下区分离神经干细胞。在有或没有抑制CK2的情况下，研究神经球的增殖及其分化。通过计数神经球的数量和测量其直径来评估增殖的变化。此外，还估计了神经球内的绝对细胞数。在神经球解离后，用视黄酸诱导单细胞分化。在分化过程诱导后的连续时间点抑制CK2。
CK2抑制剂量依赖性地减少了增殖神经球的数量和大小。在分化开始时添加CK2抑制剂CX-4945，我们观察到CX-4945对细胞活力和神经胶质细胞分化有剂量依赖性影响。在分化开始时添加第二种CK2抑制剂喹哪嗪会导致凋亡水平升高，同时神经分化减少。在分化开始72小时后添加CK2抑制剂对干细胞分化没有影响。结论：抑制CK2以时间点和浓度依赖性方式影响早期神经胶质细胞生成。
使用CK2抑制剂会显著影响神经干细胞生态位。
