Britto L R, Rogers S W, Hamassaki-Britto D E, Duvoisin R M
Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, Brazil.
Vis Neurosci. 1994 May-Jun;11(3):569-77. doi: 10.1017/s0952523800002479.
Immunohistochemical and in situ hybridization techniques were used to localize the beta 4 subunit of the neuronal nicotinic acetylcholine receptors (nAChRs) in the ground squirrel retina. The beta 4 nAChR subunit was detected in both transverse and horizontal sections of the retina using a subunit-specific antiserum and the avidin-biotin complex technique. Two bands of labeled processes were seen in the inner plexiform layer, corresponding approximately to the laminae where the cholinergic cells arborize. Labeled cells were found in the ganglion cell layer and the inner third of the inner nuclear layer. The cells in the ganglion cell layer were medium- to large-sized and were frequently observed to give rise to axon-like processes. Most of the labeled neurons in the inner nuclear layer were small presumptive amacrine cells, but a few medium-to-large cells were also labeled. These could constitute a different class of amacrine cells or displaced ganglion cells. The latter possibility is supported by the existence of nAChR-containing displaced ganglion cells in the avian retina. In situ hybridization with a 35S-labeled cRNA probe revealed the expression of mRNA coding for the nAChR beta 4 subunit in the ganglion cell layer and the inner third of the inner nuclear layer. This finding confirmed the immunohistochemical data of the cellular localization of beta 4 nAChR subunit. These results indicate that the beta 4 nAChR subunit is expressed by specific subtypes of neurons on the ground squirrel retina.(ABSTRACT TRUNCATED AT 250 WORDS)
免疫组织化学和原位杂交技术被用于在地松鼠视网膜中定位神经元烟碱型乙酰胆碱受体(nAChRs)的β4亚基。使用亚基特异性抗血清和抗生物素蛋白-生物素复合物技术,在视网膜的横向和水平切片中均检测到了β4 nAChR亚基。在内网状层可见两条标记的神经突带,大致对应于胆碱能细胞分支的板层。在神经节细胞层和内核层的内三分之一处发现了标记细胞。神经节细胞层中的细胞为中型到大型,经常观察到它们发出轴突样神经突。内核层中大多数标记神经元是假定的小无长突细胞,但也有一些中型到大型细胞被标记。这些细胞可能构成不同类型的无长突细胞或移位的神经节细胞。鸟类视网膜中存在含nAChR的移位神经节细胞支持了后一种可能性。用35S标记的cRNA探针进行原位杂交显示,神经节细胞层和内核层的内三分之一处表达了编码nAChRβ4亚基的mRNA。这一发现证实了β4 nAChR亚基细胞定位的免疫组织化学数据。这些结果表明,β4 nAChR亚基由地松鼠视网膜上特定亚型的神经元表达。(摘要截短于250字)