Zhu B S, Straznicky C, Gibbins I
Department of Anatomy and Histology, Flinders University of South Australia, Adelaide.
Vis Neurosci. 1995 Jan-Feb;12(1):11-9. doi: 10.1017/s0952523800007276.
The synaptic connections of amacrine cells synthesizing or accumulating serotonin in the retina of the cane toad, Bufo marinus, were studied by using preembedding double-labeling electron-microscopic immunocytochemistry. The binding sites of an anti-serotonin antibody were revealed by the diaminobenzidine reaction, whilst a colloidal gold-conjugated secondary antibody was used to detect an antibody to phenylalanine hydroxylase. Since the latter antibody recognizes tryptophan 5-hydroxylase, one of the synthesizing enzymes for serotonin, as well as tyrosine hydroxylase, the rate-limiting enzyme for catecholamine synthesis, the double labeling of the present study enabled us to identify three groups of labeled profiles at the ultrastructural level. The profiles of serotonin-synthesizing amacrine cells contained both diaminobenzidine reaction product and colloidal gold particles, whilst those of serotonin-accumulating and dopaminergic amacrine cells contained only diaminobenzidine reaction product or colloidal gold particles, respectively. The synapses of serotonin-synthesizing or serotonin-accumulating amacrine cells were distributed all through the inner plexiform layer of the retina. The profiles of serotonin-synthesizing amacrine cells predominantly received synapses from, and made synapses onto, unlabeled amacrine cell dendrites. They also received synapses from, and made synapses onto, bipolar cell terminals. They also made synapses onto presumed ganglion cell dendrites. However, the profiles of serotonin-accumulating cells made synapses only with unlabeled amacrine cell processes. There were close contacts between the profiles of serotonin-synthesizing and serotonin-accumulating amacrine cells. No synaptic relationships were observed between dopaminergic and serotonin-synthesizing or serotonin-accumulating amacrine cells.(ABSTRACT TRUNCATED AT 250 WORDS)
利用包埋前双标记电子显微镜免疫细胞化学技术,研究了海蟾蜍(Bufo marinus)视网膜中合成或积累5-羟色胺的无长突细胞的突触连接。抗5-羟色胺抗体的结合位点通过二氨基联苯胺反应显示,而胶体金偶联的二抗用于检测苯丙氨酸羟化酶抗体。由于后一种抗体识别5-羟色胺的合成酶之一色氨酸5-羟化酶以及儿茶酚胺合成的限速酶酪氨酸羟化酶,本研究的双标记使我们能够在超微结构水平上识别出三组标记的细胞轮廓。合成5-羟色胺的无长突细胞的轮廓同时含有二氨基联苯胺反应产物和胶体金颗粒,而积累5-羟色胺和多巴胺能无长突细胞的轮廓分别仅含有二氨基联苯胺反应产物或胶体金颗粒。合成或积累5-羟色胺的无长突细胞的突触分布在视网膜的整个内网层。合成5-羟色胺的无长突细胞的轮廓主要接受来自未标记无长突细胞树突的突触,并与它们形成突触。它们还接受来自双极细胞终末的突触,并与它们形成突触。它们也与假定的神经节细胞树突形成突触。然而,积累5-羟色胺的细胞轮廓仅与未标记的无长突细胞突起形成突触。合成5-羟色胺和积累5-羟色胺的无长突细胞轮廓之间有紧密接触。未观察到多巴胺能与合成或积累5-羟色胺的无长突细胞之间的突触关系。(摘要截短于250字)