Expression Of Nerve Growth Guidance Cue Slit2in Astrocytes And Neural Stem Cells

To investigate expression patterns of the nerve growth guidance cue slit2and its receptor robol in rat cortex astrocyters with different severity of oxygen-glucose deprivation(OGD) and in neural stem cells from the hippocampus during the process of differentiation.Methods:Cultured cortex astrocytes were designed into five groups:OGD Oh group(control group)、2h group、4h group、6h group and12h group. Morphological changes were observed by phase-contrast microscopy. MTT analysis was used to detect the cell viability. Reverse transcriptase-polymerase chain reaction (RT-PCR) was contributed to detect slit2and robol mRNA expressions.Cultured neural stem cells were designed into five groups according to the time after differentiation:Od group(control group)、1d group、2d group、3d group and7d group. Immunofluorescence stain was used to identify neural stem cells、neurons and astrocytes.RT-PCR was contributed to detect slit2and robol mRNA expressions.Results:(1)The cell activities(OD values) were (0.756±0.013),(0.681±0.016),(0.563±0.033),(0.404±0.024) and (0.222±0.026) in OGD Oh group、2h group、4h group、6h group and12h group respectively(p<0.01),with time-related manner.(2)After oxygen-glucose deprivation,statistics showed that the expressions of slit2and robol mRNA in astrocytes were first up-regulated,peaked at6h(p<0.05,compared with control group),and then decreased.Compared with6h group,both of the expressions were decreased significantly in12h group(both p<0.01).(3)Cultured neural stem cells had the ability of self proliferation self renewal and differentiating into neurons and astrocytes.(4)Expressions of robol mRNA during the differentiation of neural stem cells:①There were no significant differences between the expressions of control group、1d group and2d group(p>0.05)②The expression of3d group was higher than that of control group、1d group and2d group (p<0.05)③The expression of7d group was much higher than that of control group、1d group and2d group (p<0.01), but not significantly higher than3d group (p>0.05), even though rised.(5)Expression of slit2mRNA during the differentiation of neural stem cells:there were no significant differences among the expressions of all groups(p>0.05).Conclusion:The expressions of slit2/robol can be up-regulated either in astrocytes after OGD or in neural stem cells during the differentiaion, suggesting that slit2/robol may be involved in the regulation of neural regeneration.