Thorogood M S, Almeida V W, Brodfuehrer P D
Biology Department, Bryn Mawr College, Pennsylvania 19010, USA.
J Comp Neurol. 1999 Mar 15;405(3):334-44. doi: 10.1002/(sici)1096-9861(19990315)405:3<334::aid-cne4>3.0.co;2-i.
Previous physiological and pharmacological evidence has suggested a neurotransmitter role for the excitatory amino acid glutamate in the leech central nervous system (CNS). In the present study, we sought to localize glutamate receptor (GluR) subunits (GluR 5/6/7, GluR 2/3 and N-methyl-D-aspartate receptor 1 [NMDAR 1]) and a glutamate transporter subtype [GLT-1] within the leech CNS using mono- and polyclonal antibodies. In whole-mounted tissue, small cells of the outer capsule and putative microglia labeled with both GluR 5/6/7 and GluR 2/3 but not NMDAR 1 subunit antisera. In general, GluR 5/6/7-like immunofluorescence was both more intense and more widespread than GluR 2/3-like immunolabeling. Cryostat-sectioned tissue revealed extensive GluR 5/6/7-like immunoreactivity throughout the neuropil as well as labeling within a few neuronal somata. GLT-1-like immunoreactivity localized to the inner capsule, which is the interface between neuronal somata and the neuropil and is deeply invested by processes of neuropil glia. These results complement previous physiological and pharmacological findings indicating that the leech CNS possesses the cellular machinery to respond to glutamate and to transport glutamate from extracellular spaces. Together, they provide further evidence for glutamate's role as a neurotransmitter within the leech CNS.
先前的生理学和药理学证据表明,兴奋性氨基酸谷氨酸在水蛭中枢神经系统(CNS)中发挥神经递质的作用。在本研究中,我们试图使用单克隆抗体和多克隆抗体在水蛭中枢神经系统中定位谷氨酸受体(GluR)亚基(GluR 5/6/7、GluR 2/3和N-甲基-D-天冬氨酸受体1 [NMDAR 1])以及一种谷氨酸转运体亚型[GLT-1]。在整装组织中,外囊的小细胞和假定的小胶质细胞被GluR 5/6/7和GluR 2/3抗体标记,但未被NMDAR 1亚基抗血清标记。一般来说,GluR 5/6/7样免疫荧光比GluR 2/3样免疫标记更强且分布更广。冰冻切片组织显示,在整个神经纤维网中都有广泛的GluR 5/6/7样免疫反应性,并且在一些神经元胞体中也有标记。GLT-1样免疫反应性定位于内囊,内囊是神经元胞体与神经纤维网之间的界面,并且被神经纤维网胶质细胞的突起深深包裹。这些结果补充了先前的生理学和药理学发现,表明水蛭中枢神经系统具备对谷氨酸做出反应并从细胞外空间转运谷氨酸的细胞机制。它们共同为谷氨酸在水蛭中枢神经系统中作为神经递质的作用提供了进一步的证据。