Brunken W J, Witkovsky P, Karten H J
J Comp Neurol. 1986 Jan 1;243(1):1-12. doi: 10.1002/cne.902430102.
We surveyed retinas of Raja erinacea, Mustelus canis, and Squalus acanthias for neurotransmitter substances by using antisera directed against the substances themselves or against their synthesizing enzymes. Both the peroxidase-antiperoxidase (PAP) and indirect fluorescent techniques were employed to visualize the primary antisera. In all three species positive results were obtained with antisera directed against tyrosine hydroxylase (TOH), glutamic acid decarboxylase (GAD), serotonin (5-HT), and leucine enkephalin (Lenk). Antisera directed against glucagon, neurotensin, beta-endorphin, vasoactive intestinal peptide, or bombesin failed to show any specific staining. Immunoreactivity was located in amacrine, interplexiform, and horizontal cells as well as in axons of the optic fiber layer. The four antisera labelled different amacrine cell classes, distinguished on the bases of perikaryal morphology and the distribution of cell processes in the inner plexiform layer (IPL). Amacrine cells that labelled with the same marker were seen to have different morphologies in the species studied. Thus, TOH-like immunoreactivity was distributed in layers 1, 3, and 5 of the IPL in Mustelus but only in layers 1 and 3 in Raja retina. GAD-like immunoreactivity was found diffusely over all layers of the IPL in Raja, but in Mustelus it was confined primarily to layers 1, 3, and 5 of the IPL. Lenk- and 5-HT-like immunoreactivities showed similar species variations. Two neurochemical classes of interplexiform cell were identified in this study. In Mustelus GAD-like and Lenk-like immunoreactive interplexiform cells were seen whereas in Raja only GAD-positive interplexiform cells were detected. In squalus no unequivocal demonstration of any interplexiform cell was made with these antisera. The GAD antiserum also labelled a subset of the horizontal cells in the dorsal retina of Raja. TOH and 5-HT-antisera labelled axons in the optic fiber layer of all three species but reactive ganglion cell perikarya were not identified.
我们通过使用针对神经递质物质本身或其合成酶的抗血清,对猬鳐、犬鲨和棘鲨的视网膜进行了神经递质物质的检测。采用过氧化物酶-抗过氧化物酶(PAP)和间接荧光技术来显示一抗。在所有这三个物种中,针对酪氨酸羟化酶(TOH)、谷氨酸脱羧酶(GAD)、5-羟色胺(5-HT)和亮氨酸脑啡肽(Lenk)的抗血清都得到了阳性结果。针对胰高血糖素、神经降压素、β-内啡肽、血管活性肠肽或蛙皮素的抗血清未显示任何特异性染色。免疫反应性位于无长突细胞、网间细胞和水平细胞以及视神经纤维层的轴突中。这四种抗血清标记了不同类别的无长突细胞,这些细胞类别是根据胞体形态和内网状层(IPL)中细胞突起的分布来区分的。在所研究的物种中,用相同标记物标记的无长突细胞具有不同的形态。因此,在犬鲨中,TOH样免疫反应性分布在IPL的第1、3和5层,但在猬鳐视网膜中仅分布在第1和3层。在猬鳐中,GAD样免疫反应性在IPL的所有层中广泛分布,但在犬鲨中,它主要局限于IPL的第1、3和5层。Lenk样和5-HT样免疫反应性表现出类似的物种差异。在本研究中鉴定出了两类神经化学性质的网间细胞。在犬鲨中可以看到GAD样和Lenk样免疫反应性网间细胞,而在猬鳐中仅检测到GAD阳性网间细胞。在棘鲨中,用这些抗血清未能明确显示任何网间细胞。GAD抗血清还标记了猬鳐背侧视网膜中一部分水平细胞。TOH和5-HT抗血清标记了所有三个物种视神经纤维层中的轴突,但未识别出有反应的神经节细胞胞体。