Townes-Anderson E, Vogt B A
Department of Physiology and Biophysics, Cornell University Medical College, New York, New York 10021.
J Comp Neurol. 1989 Dec 15;290(3):369-83. doi: 10.1002/cne.902900306.
Binding of propylbenzilylcholine mustard, a muscarinic acetylcholine receptor antagonist, to isolated retinal cells was examined with light microscopic autoradiography. Dissociation of the adult tiger salamander retina yielded identifiable rod, cone, horizontal, bipolar, amacrine/ganglion, and Müller cells. Preservation of fine structure was assessed with conventional electron microscopy. For all cell types, the plasmalemma was intact and free of adhering debris; in addition, presynaptic ribbon complexes were present in photoreceptor and bipolar axon terminals indicating that synaptic structures were retained. Specific binding to cell bodies and processes was analyzed separately by using morphometric and statistical techniques. The highest grain densities occurred on processes of amacrine/ganglion cells and axons and 2 degrees and 3 degrees dendrites of bipolar neurons. Bipolar cells, however, seemed to be a heterogeneous population because there was great variation in the density of binding sites on both their axons and distal dendrites. Intermediate levels of binding were found on bipolar 1 degree dendrites and horizontal cells. No specific binding was detected on Müller cells and most parts of photoreceptors. Comparisons between cells showed that grain densities were similar for bipolar axons and amacrine/ganglion cell processes but bipolar dendrites were richer in binding sites than horizontal cell dendrites. Thus, muscarinic receptors in the salamander retina are located on amacrine/ganglion, bipolar, and horizontal cells and primarily confined to the processes which compose the two synaptic layers. In the inner plexiform layer, muscarinic receptors reside on processes from all three inner retinal neurons: in the outer synaptic layer, receptors are only on second-order cells and are more numerous along bipolar than horizontal cell dendrites.
利用光学显微镜放射自显影技术,检测了毒蕈碱型乙酰胆碱受体拮抗剂丙基苯甲酰胆碱氮芥与分离的视网膜细胞的结合情况。成年虎蝾螈视网膜解离后可得到可识别的视杆细胞、视锥细胞、水平细胞、双极细胞、无长突/神经节细胞和米勒细胞。用传统电子显微镜评估精细结构的保存情况。对于所有细胞类型,质膜完整且无附着碎片;此外,在光感受器和双极轴突终末存在突触前带复合体,表明突触结构得以保留。通过形态计量学和统计技术分别分析了与细胞体和突起的特异性结合。最高的颗粒密度出现在无长突/神经节细胞的突起、轴突以及双极神经元的二级和三级树突上。然而,双极细胞似乎是一个异质群体,因为它们的轴突和远端树突上结合位点的密度存在很大差异。在双极一级树突和水平细胞上发现了中等水平的结合。在米勒细胞和大部分光感受器上未检测到特异性结合。细胞间比较显示,双极轴突和无长突/神经节细胞突起上的颗粒密度相似,但双极树突上的结合位点比水平细胞树突上的更丰富。因此,蝾螈视网膜中的毒蕈碱受体位于无长突/神经节细胞、双极细胞和水平细胞上,主要局限于构成两个突触层的突起上。在内网状层,毒蕈碱受体存在于所有三个视网膜内层神经元的突起上;在外突触层,受体仅存在于二级细胞上,并且沿双极细胞树突的数量比水平细胞树突上的更多。