Study On The Role Of Divalent Metal Transporter DMT1 In Neural Stem Cells

As an important biological phenomenon in the mammalian brain,neurogenesis is a process by which neural stem cells produce mature and highly differentiated functional nerve cells,which are critical to the development and maturation of the nervous system,it’s also closely related to advanced brain functions such as learning and memory.Neural stem cells have a pivotal position in the process of neurogenesis,it is affected by a number of factors as it functions,for example: physiological factors and local microenvironment,are of great significance to the development and pathogenesis of the nervous system.However,research on the roles of DMT1 on neural stem cells is not clear.As a metal transporter,DMT1 plays an important role in the body,studies have found that due to its ability to transport metal ions,it can lead to the deposition of some metal ions in the brain,resulting in serious neurological diseases.However,there have been no reports on the association between DMT1 and neural stem cells.Based on the model of manganese poisoning,this study explored the role of DMT1 in neural stem cells,providing new ideas for the treatment and pathogenesis of neurodevelopmental diseases and providing a basis for clinical work.We cultured the neural stem cells,neuron and astrocyte,the expression level of DMT1 protein was detected by Western Blot;RNA was extracted from the organs,brain regions and developmental brain of eight-week-old male C57BL/6 mice,the expression of DMT1 was studied by PCR experiments.Compared with other organs,DMT1 was highly expressed in the cerebral cortex,but no significant difference in expression in each brain region;The expression of DMT1 in the mouse brain increased from the 14 th day of embryo,and decreased after the peak of P0,and then stabilized in adulthood.This suggests that DMT1 may play an important role in the developing brain of mice.We cultured the neural stem cells,neuron and astrocyte,and the expression level of DMT1 protein was detected by Western Blot.We found that DMT1 expression in neural stem cell was higher than that in neuron and astrocyte.To further explore the role of DMT1 in neural stem cells,we constructed the DMT1 overexpressing virus and infected neural stem cells in vitro.It was found that the proliferation of neural stem cells decreased after overexpressing DMT1.This may be related to the transport of DMT1 to metal ions in neural stem cells.Overexpression DMT1 may cause the deposition of metal ions in neural stem cells,which in turn reduces the proliferation of neural stem cells.For further investigation,our project were based on the manganese exposure model.We cultured the neural stem cells and treated them with different concentrations of manganese(100 μM and 300 μM),after 24 h we compared the proliferation of neural stem cells and found that manganese exposure group could decrease the proliferation of neural stem cells,as the concentration of manganese increases,the proliferation of neural stem cells is more significantly inhibited.Meanwhile,we extracted the protein and found that DMT1 expression in the manganese group was significantly decreased compared with that in the control group.Furthermore,we constructed the manganese exposure models and the DG was extracted using western blot to detect the DMT1 expression.Western blot showed that DMT1 expression of neural stem cells in the manganese group was significantly decreased compared with that in the control group.This showed that DMT1 may play a role in transport during manganese exposure of neural stem cells.As the concentration of manganese increases,the expression of DMT1 decreases to achieve a reduced transport of manganese,which may be the protective effect of DMT1 on neural stem cells.It is suggested that with the increase of manganese concentration,the expression of DMT1 decreases to reduce the transport of manganese,which may be the protective effect of DMT1 on neural stem cells.In conclusion,phenotypic analysis showed that the expression of DMT1 is increased during brain development,and the expression in neural stem cells is higher than that in neuron and astrocyte.After overexpression of DMT1,the proliferation ability of neural stem cells is weakened.We used neural stem cells with manganese exposure as models and found that not only was the proliferation,but the expression of DMT1 was also decreased;DMT1 expression was also significantly decreased in the mouse manganese exposure model.DMT1 may play a protective role in neural stem cells during this process.They all confirmed this conclusion.Therefore,DMT1 plays an important role in manganese exposure and proliferation of neural stem cells.This provides a theoretical basis for the research and clinical work of manganese exposure mechanism,as well as novel idea for the treatment and pathogenesis of neurodevelopment-related diseases.