Laboratory of Neuroscience, Department of Biophysics, Wroclaw Medical University, Wroclaw, Poland.
J Physiol Pharmacol. 2010 Aug;61(4):501-6.
Acute hippocampal slices and primary neuronal cultures are often used with a tacit assumption that basic characteristics of the two models closely resemble each other. The use of the cell cultures, however, may raise controversies because of non-physiological conditions resulting from e.g. glial cells deficit, random neuronal sprouting, lack of specificity in the synaptic connections, impaired homeostasis, etc. Importantly, alteration in neuronal environment, especially when occurring over a prolonged period of time, may give rise to a profound homeostatic modulation. In the present study we have compared the properties of GABAergic and glutamatergic (non-NMDA) currents in pyramidal neurons from hippocampal slices and neuronal cell culture. We show that, most strikingly, amplitude ratio of currents elicited by ultrafast applications of saturating GABA and glutamate was nearly one order of magnitude larger in cultured neurons than that in slices. Miniature IPSCs and EPSCs also showed substantial differences between these two models. In particular, mEPSC amplitudes were larger and more frequent in cultured neurons but their time duration was longer in slices. Miniature IPSCs did not show differences in amplitude when comparing slices and cultures but their time duration was faster and occurrence more frequent in slices. In conclusion, we provide evidence that expression pattern of GABA(A) and glutamate receptors as well as synaptic current properties in the neuronal cell culture show profound differences with respect to that in the physiological conditions.
急性海马脑片和原代神经元培养物常被用于研究,其隐含假设是这两种模型的基本特征非常相似。然而,由于例如神经胶质细胞缺乏、随机神经元突增、突触连接缺乏特异性、内稳态受损等非生理条件的存在,细胞培养物的使用可能会引发争议。重要的是,神经元环境的改变,特别是当这种改变持续很长时间时,可能会引起深刻的内稳态调节。在本研究中,我们比较了海马脑片和神经元细胞培养物中锥体细胞的 GABA 能和谷氨酸能(非 NMDA)电流的特性。我们发现,最显著的是,在培养神经元中,超快应用饱和 GABA 和谷氨酸诱发的电流的幅度比值比在脑片中几乎大一个数量级。微小 IPSC 和 EPSC 也在这两种模型之间显示出显著差异。特别是,培养神经元中的 mEPSC 幅度较大且更频繁,但持续时间较长。在比较脑片和培养物时,微小 IPSC 的幅度没有差异,但持续时间更快,发生频率更高。总之,我们提供的证据表明,神经元细胞培养物中 GABA(A)和谷氨酸受体的表达模式以及突触电流特性与生理条件下存在显著差异。