Differential Cholinergic Phenotypes in Primary Culture Neurons and Brain In Vivo
Li Yimei, Wang Guoxiang, Wang Yun*
To further understand whether primary cultured cholinergic neurons can be used as a viable cell model for cholinergic neurons in vitro, we compared differences in proportion of neurons expressed cholinergic biomarkers and total neurons in cultured neurons cultivated from E18 basal forebrain (BF) and E18 hippocampus (HIP), and the differences in expression ratio of cholinergic markers between primary culture neurons and brain slices in the same brain regions. Immunofluorescence was used to detect the proportion of ChAT and p75NTR (two commonly used cholinergic neuron markers) expression in DIV 3 and DIV 21 of cultured neurons of basal forebrain and hippocampus from E18 fetus, and also in E18 fetus and adult mice in brain slice. Both ChAT and p75NTR showed high expression ratio in cultured neurons from BF and HIP in DIV 3 and DIV 21. However, there was no expression of ChAT in hippocampal slices of E18 fetus and adult mice. Therefore, the ratio of neurons expressed cholinergic markers in cultured neurons from basal forebrain and hippocampus is inconsistent with in vivo, suggesting that it is temporarily not feasible to culture primary cholinergic neurons in vitro by using primary neuronal culture methods. Meanwhile, in addition to immunostaining more techniques and methods are needed to identify cholinergic neurons in cultured cells.
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