Doucet Edith, Latrémolière Alban, Darmon Michèle, Hamon Michel, Emerit Michel B
INSERM U677, F-75013 Paris, France.
Eur J Neurosci. 2007 Jul;26(2):355-66. doi: 10.1111/j.1460-9568.2007.05659.x.
The 5-HT(3) receptor is a member of the superfamily of neurotransmitter-gated ion channels involved in fast synaptic signalling and in modulation of neurotransmitter release. As for many other channel receptors, the electrophysiological properties and the functions of the 5-HT(3) receptor are determined by subunit composition of the pentameric channel. Because in situ hybridization did not allow the detection of mRNA encoding the 5-HT(3B) subunit in the rodent central nervous system, or in nearly half of the neurons expressing the 5-HT(3A) subunit in peripheral ganglia, it has been suggested that subunit composition could define at least two 5-HT(3) receptor-expressing neuronal populations. In order to challenge this hypothesis, we have developed polyclonal antibodies directed against a portion of the second intracytoplasmic loop of the mouse 5-HT(3B) subunit. Immunohistochemical analysis in the mouse and the rat revealed that immunolabelling was most prominent in peripheral ganglia, particularly in trigeminal ganglia (TG). In rats, transection or ligature of the infraorbital nerve resulted in a pronounced accumulation of immunoreactive material at the proximal side of the lesioned nerve, and an up-regulation of both subunits in 5-HT(3) receptor-expressing TG neurons. Surprisingly, nearly 100% of neurons expressing 5-HT(3A) subunits were also labelled by anti-5-HT(3B) antibodies. We also detected 5-HT(3B) immunoreactivity in the rat hippocampal CA1 layer and in scattered cortical neurons, indicating that detection of 5-HT(3) subunit mRNA by in situ hybridization might not provide really complete mapping of heteromeric 5-HT(3A/B) vs. homomeric 5-HT(3A) receptors in the peripheral and central nervous systems in rodents.
5-羟色胺(3)受体是神经递质门控离子通道超家族的成员,参与快速突触信号传递和神经递质释放的调节。与许多其他通道受体一样,5-羟色胺(3)受体的电生理特性和功能由五聚体通道的亚基组成决定。由于原位杂交无法在啮齿动物中枢神经系统中检测到编码5-羟色胺(3B)亚基的mRNA,在外周神经节中表达5-羟色胺(3A)亚基的近半数神经元中也检测不到,因此有人提出亚基组成可能至少定义了两个表达5-羟色胺(3)受体的神经元群体。为了验证这一假设,我们制备了针对小鼠5-羟色胺(3B)亚基第二个胞质内环部分的多克隆抗体。对小鼠和大鼠的免疫组织化学分析显示,免疫标记在外周神经节中最为明显,尤其是在三叉神经节(TG)中。在大鼠中,眶下神经横断或结扎导致损伤神经近端侧免疫反应性物质明显积聚,以及表达5-羟色胺(3)受体的TG神经元中两个亚基均上调。令人惊讶的是,几乎100%表达5-羟色胺(3A)亚基的神经元也被抗5-羟色胺(3B)抗体标记。我们还在大鼠海马CA1层和散在的皮质神经元中检测到5-羟色胺(3B)免疫反应性,这表明通过原位杂交检测5-羟色胺(3)亚基mRNA可能无法真正完整地绘制啮齿动物外周和中枢神经系统中异源5-羟色胺(3A/B)与同源5-羟色胺(3A)受体的分布图。