Department of Biology, University of Utah Salt Lake City, UT, USA.
Department of Biology, University of Utah Salt Lake City, UT, USA ; Interdepartmental Neuroscience Program, University of Utah Salt Lake City, UT, USA.
Front Cell Neurosci. 2013 Nov 28;7:225. doi: 10.3389/fncel.2013.00225. eCollection 2013.
We investigated the functional expression of nicotinic acetylcholine receptors (nAChRs) in heterogeneous populations of dissociated rat and mouse lumbar dorsal root ganglion (DRG) neurons by calcium imaging. By this experimental approach, it is possible to investigate the functional expression of multiple receptor and ion-channel subtypes across more than 100 neuronal and glial cells simultaneously. Based on nAChR expression, DRG neurons could be divided into four subclasses: (1) neurons that express predominantly α3β4 and α6β4 nAChRs; (2) neurons that express predominantly α7 nAChRs; (3) neurons that express a combination of α3β4/α6β4 and α7 nAChRs; and (4) neurons that do not express nAChRs. In this comparative study, the same four neuronal subclasses were observed in mouse and rat DRG. However, the expression frequency differed between species: substantially more rat DRG neurons were in the first three subclasses than mouse DRG neurons, at all developmental time points tested in our study. Approximately 70-80% of rat DRG neurons expressed functional nAChRs, in contrast to only ~15-30% of mouse DRG neurons. Our study also demonstrated functional coupling between nAChRs, voltage-gated calcium channels, and mitochondrial Ca(2) (+) transport in discrete subsets of DRG neurons. In contrast to the expression of nAChRs in DRG neurons, we demonstrated that a subset of non-neuronal DRG cells expressed muscarinic acetylcholine receptors and not nAChRs. The general approach to comparative cellular neurobiology outlined in this paper has the potential to better integrate molecular and systems neuroscience by uncovering the spectrum of neuronal subclasses present in a given cell population and the functionally integrated signaling components expressed in each subclass.
我们通过钙成像技术研究了分离的大鼠和小鼠腰背部脊神经节(DRG)神经元异质群体中烟碱型乙酰胆碱受体(nAChRs)的功能表达。通过这种实验方法,可以同时研究超过 100 个神经元和神经胶质细胞中多种受体和离子通道亚型的功能表达。根据 nAChR 的表达情况,DRG 神经元可分为四个亚类:(1)主要表达α3β4 和α6β4 nAChRs 的神经元;(2)主要表达α7 nAChRs 的神经元;(3)同时表达α3β4/α6β4 和α7 nAChRs 的神经元;(4)不表达 nAChRs 的神经元。在这项比较研究中,在小鼠和大鼠的 DRG 中观察到了相同的四个神经元亚类。然而,表达频率在不同物种之间存在差异:在我们研究的所有发育时间点,大鼠 DRG 神经元中属于前三个亚类的比例明显高于小鼠 DRG 神经元。大约 70-80%的大鼠 DRG 神经元表达功能性 nAChRs,而小鼠 DRG 神经元中只有约 15-30%表达。我们的研究还证明了 nAChRs、电压门控钙通道和离散 DRG 神经元亚群中线粒体 Ca(2) (+) 转运之间的功能偶联。与 DRG 神经元中 nAChRs 的表达相反,我们证明了一小部分非神经元 DRG 细胞表达毒蕈碱型乙酰胆碱受体而不是 nAChRs。本文概述的比较细胞神经生物学的一般方法有可能通过揭示给定细胞群体中存在的神经元亚类的范围以及每个亚类中表达的功能整合信号成分,更好地整合分子和系统神经科学。