Dallas Mark, Deuchars Susan A, Deuchars Jim
School of Biomedical Sciences, University of Leeds, Leeds LS2 9JT, UK.
J Neurosci Methods. 2005 Aug 15;146(2):133-48. doi: 10.1016/j.jneumeth.2005.04.015.
The application of antibodies to living neurones has the potential to modulate function of specific proteins by virtue of their high specificity. This specificity has proven effective in determining the involvement of many proteins in neuronal function where specific agonists and antagonists do not exist, e.g. ion channel subunits. We discuss studies where antibodies modulate functions of voltage gated sodium, voltage gated potassium, voltage gated calcium hyperpolarisation activated cyclic nucleotide (HCN gated) and transient receptor potential (TRP) channels. Ligand gated channels studied in this way include nicotinic acetylcholine receptors, purinoceptors and GABA receptors. Antibodies have also helped reveal the involvement of different intracellular proteins in neuronal functions including G-proteins as well as other proteins involved in trafficking, phosphoinositide signalling and neurotransmitter release. Some suggestions for control experiments are made to help validate the method. We conclude that antibodies can be extremely valuable in determining the functions of specific proteins in living neurones in neuroscience research.
将抗体应用于活神经元有潜力凭借其高特异性调节特定蛋白质的功能。这种特异性已被证明在确定许多不存在特异性激动剂和拮抗剂的蛋白质参与神经元功能方面是有效的，例如离子通道亚基。我们讨论了抗体调节电压门控钠通道、电压门控钾通道、电压门控钙通道、超极化激活环核苷酸（HCN门控）通道和瞬时受体电位（TRP）通道功能的研究。以这种方式研究的配体门控通道包括烟碱型乙酰胆碱受体、嘌呤受体和GABA受体。抗体还有助于揭示不同细胞内蛋白质参与神经元功能，包括G蛋白以及参与转运、磷酸肌醇信号传导和神经递质释放的其他蛋白质。文中提出了一些对照实验的建议，以帮助验证该方法。我们得出结论，在神经科学研究中，抗体对于确定活神经元中特定蛋白质的功能可能极其有价值。
