Ríos J D, Forde K, Diebold Y, Lightman J, Zieske J D, Dartt D A
Schepens Eye Research Institute and Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts 02114, USA.
Invest Ophthalmol Vis Sci. 2000 Jul;41(8):2127-37.
To investigate expression of muscarinic, cholinergic, and adrenergic receptors on developing conjunctival goblet cells.
Eyes were removed from rats 9 to 60 days old, fixed, and used for microscopy. For glycoconjugate expression, sections were stained with Alcian blue/periodic acid-Schiffs reagent (AB/PAS) and with the lectins Ulex europeus agglutinin I (UEA-I) and Helix pomatia agglutinin (HPA). Goblet cell bodies were identified using anti-cytokeratin 7 (CK7). Nerve fibers were localized using anti-protein gene product 9.5. Location of muscarinic and adrenergic receptors was investigated using anti-muscarinic and beta-adrenergic receptors.
At days 9 and 13, single apical cells in conjunctival epithelium stained with AB/PAS, UEA-I, and CK7. At days 17 and 60, increasing numbers of goblet cells were identified by AB/PAS, UEA-I, HPA, and CK7. Nerve fibers were localized around stratified squamous cells and at the epithelial base at days 9 and 13, and around goblet cells and at the epithelial base at days 17 and 60. At days 9 and 13, M2- and M3-muscarinic and beta2-adrenergic receptors were found in stratified squamous cells, but M1-muscarinic and beta1-adrenergic receptors were not detected. At days 17 and 60, M2- and M3-muscarinic receptors were found in goblet cells, whereas M1-muscarinic receptors were in stratified squamous cells. Beta1- and beta2-adrenergic receptors were found on both cell types. Beta3-adrenergic receptors were not detected.
In conjunctiva, nerves, M2- and M3-muscarinic, and beta1- and beta2-adrenergic receptors are present on developing goblet cells and could regulate secretion as eyelids open.
研究毒蕈碱、胆碱能和肾上腺素能受体在发育中的结膜杯状细胞上的表达。
从9至60日龄大鼠摘除眼球,固定后用于显微镜检查。对于糖缀合物表达,切片用阿尔辛蓝/过碘酸希夫试剂(AB/PAS)以及凝集素欧洲荆豆凝集素I(UEA-I)和苹果蜗牛凝集素(HPA)染色。使用抗细胞角蛋白7(CK7)鉴定杯状细胞体。使用抗蛋白基因产物9.5定位神经纤维。使用抗毒蕈碱和β-肾上腺素能受体研究毒蕈碱和肾上腺素能受体的位置。
在第9天和第13天,结膜上皮中的单个顶端细胞用AB/PAS、UEA-I和CK7染色。在第17天和第60天,通过AB/PAS、UEA-I、HPA和CK7鉴定出越来越多的杯状细胞。在第9天和第13天,神经纤维位于复层鳞状细胞周围和上皮基部,在第17天和第60天位于杯状细胞周围和上皮基部。在第9天和第13天,在复层鳞状细胞中发现M2和M3毒蕈碱受体以及β2肾上腺素能受体,但未检测到M1毒蕈碱受体和β1肾上腺素能受体。在第17天和第60天,在杯状细胞中发现M2和M3毒蕈碱受体,而M1毒蕈碱受体存在于复层鳞状细胞中。在两种细胞类型上均发现β1和β2肾上腺素能受体。未检测到β3肾上腺素能受体。
在结膜中,神经、M2和M3毒蕈碱受体以及β1和β2肾上腺素能受体存在于发育中的杯状细胞上,并且在眼睑张开时可能调节分泌。