Sarasquete C, Gisbert E, Ribeiro L, Vieira L, Dinis M T
Instituto de Ciencias Marinas de Andalucía, CSIC, Puerto Real, Cádiz, Spain.
Eur J Histochem. 2001;45(3):267-78. doi: 10.4081/1637.
Epidermal, branchial and digestive mucous cells, and the gastric glands of larvae/postlarvae (from hatching until 45 days posthatching) of three fish species (two teleostean and a chondrostean) were investigated using conventional histochemical methods (periodic acid schiff -PAS-, diastase-PAS; alcian blue pH 0.5, 1 and 2.5) in order to distinguish neutral and acidic (carboxylated and sulphated) glycoconjugates, as well as bromophenol blue reaction for identification of proteins. Additionally, the presence and distribution of sugar residues in the oligosaccharide side chains of glycoconjugates were investigated using horseradish peroxidase (HPR)-conjugated lectins (Con A, DBA, WGA and UEA-I). Most mucous cells (digestive, epidermal and branchial) of Siberian sturgeon, Acipenser baeri, sea bream, Sparus aurata and Senegal sole, Solea senegalensis larvae were PAS- and alcian blue- (pH 2.5 and 0.5) positive, with small variations between organs/tissues and species. Bromophenol blue reaction (general proteins) was positive in a minority of the mucous cells, usually in those cells which were PAS-negative. Proteins rich in sulphydryl (-SH) and/or disulphide (-S-S-) groups related with the glycoprotein nature of the glycoconjugates present in mucous cells were also observed. Epidermal, branchial and digestive mucous cells of all studied larvae did not contain glycogen or lipids. Con A lectin staining was negative in all mucous cells types of sea bream and sole, but oesophageal mucous cell of sturgeon were reactive to different lectin reactions, suggesting the presence of mannose -Man- and/or glucose -Glc-, L-fucose -Fuc- ; N-acetyl-D-galactosamine -GalNAc-, as well as N-acetyl-D-glucosamine- GlcNAc - and/or sialic acid -NANA- residues. Digestive mucous cells of all studied larvae were positive to WGA and DBA lectins. Epidermal and branchial mucous cells of sea bream and sole were Con A, DBA and UEA-I unreactive. However, mucous cells of sturgeon larvae were stained with UEA-I lectin. Gastric glands appear very early in sturgeon stomach larvae development (between 5-6 days posthatching) but rather late (around 40 days) during the ontogeny of sole and sea bream larvae. These glands contain neutral glycoproteins with Man and/or Glc, Fuc, GlcNAc- and/or sialic acid and rich in GalNAc- sugar residues, as well as proteins moderately rich in arginine, and others particularly rich in tyrosine and tryptophan.
采用常规组织化学方法(过碘酸希夫反应-PAS-、淀粉酶-PAS;阿尔辛蓝pH 0.5、1和2.5)对三种鱼类(两种硬骨鱼和一种软骨硬鳞鱼)幼体/后幼体(从孵化到孵化后45天)的表皮、鳃和消化黏液细胞以及胃腺进行了研究,以区分中性和酸性(羧化和硫酸化)糖缀合物,以及用于蛋白质鉴定的溴酚蓝反应。此外,使用辣根过氧化物酶(HPR)偶联凝集素(伴刀豆球蛋白A、大豆凝集素、麦胚凝集素和荆豆凝集素-I)研究了糖缀合物寡糖侧链中糖残基的存在和分布。西伯利亚鲟、欧洲海鲈和塞内加尔鳎幼体的大多数黏液细胞(消化、表皮和鳃)对PAS和阿尔辛蓝(pH 2.5和0.5)呈阳性反应,器官/组织和物种之间存在细微差异。溴酚蓝反应(一般蛋白质)在少数黏液细胞中呈阳性,通常在那些PAS阴性的细胞中。还观察到与黏液细胞中存在的糖缀合物糖蛋白性质相关的富含巯基(-SH)和/或二硫键(-S-S-)的蛋白质。所有研究幼体的表皮、鳃和消化黏液细胞均不含糖原或脂质。伴刀豆球蛋白A凝集素染色在海鲈和鳎的所有黏液细胞类型中均为阴性,但鲟鱼的食管黏液细胞对不同的凝集素反应有反应,表明存在甘露糖-Man-和/或葡萄糖-Glc-、L-岩藻糖-Fuc-;N-乙酰-D-半乳糖胺-GalNAc-,以及N-乙酰-D-葡萄糖胺-GlcNAc-和/或唾液酸-NANA-残基。所有研究幼体的消化黏液细胞对麦胚凝集素和大豆凝集素呈阳性反应。海鲈和鳎的表皮和鳃黏液细胞对伴刀豆球蛋白A、大豆凝集素和荆豆凝集素-I无反应。然而,鲟鱼幼体的黏液细胞用荆豆凝集素-I染色。胃腺在鲟鱼胃幼体发育早期(孵化后5-6天之间)出现,但在鳎和海鲈幼体的个体发育过程中出现较晚(约40天)。这些腺体含有含有Man和/或Glc、Fuc、GlcNAc-和/或唾液酸且富含GalNAc-糖残基的中性糖蛋白,以及中度富含精氨酸的蛋白质,还有特别富含酪氨酸和色氨酸的其他蛋白质。