Abstract:

To study the necessary of retinoic acid receptor α (RARα) for rat neuron function.
Methods Tissue digestion was used to isolate and cultivate the rat primary hippocampal neurons,and the adenovirus vector was used to  specifically silence the  RARα;Real-Time PCR was used to analyze the influence of silenced RARα in  retinoic acid(RA) receptors and the markers of nerve cells;live cell imaging analysis was performed to analyze the influence of the calcium excitability of neurons silenced RARα.Results The immunofluorescence results showed that  90% of the isolated cells expressed the  neuron marker neuron-specific enolase (NSE),the adenoviral transfection efficiency was up to 80%.The PCR results showed the expression of RARα in silenced RARα neuron was decreased by 75% (P<0.01),the other receptors were significantly decreased (P<0.01),but RARβ was significantly increased (P<0.05).The live cell calcium imaging results showed the calcium excitability in silent group was significantly reduced (P<0.05),however all-trans retinoic acid (ATRA) pretreatment for 24 h could significantly enhance the calcium excitability (P<0.01).Conclusion The absence of RARα can significantly reduce the neuron marker NSE expression of the primary hippocampal neurons,and significantly damage the neuronal calcium excitability.

Key words: primary hippocampal neurons；retinoic acid；retinoic acid receptor α；calcium excitability；gene silence