Pixley S K, Farbman A I, Menco B P
Department of Cell Biology, Neurobiology, and Anatomy, University of Cincinnati College of Medicine, Ohio 45267-0521, USA.
Anat Rec. 1997 Jul;248(3):307-21. doi: 10.1002/(SICI)1097-0185(199707)248:3<307::AID-AR2>3.0.CO;2-M.
The olfactory epithelial sustentacular cells may support the survival and function of olfactory receptor neurons, but few reagents are available to mark and manipulate such cells.
Novel nasal cell-specific monoclonal antibodies were generated using whole cultured rat olfactory mucosal cells as the antigenic stimuli. They were characterized by immunostaining at the light level in rat tissues and newborn rat olfactory cell cultures, and at the electron microscopic level in adult tissues using freeze-substitution, post-embedding staining.
An IgMkappa monoclonal antibody designated 1F4 selectively labeled apical surfaces of the rat olfactory and respiratory epithelia in tissue sections and what appeared to be sustentacular cells in olfactory cell cultures. Using electron microscopy, 1F4 bound selectively to the microvilli of sustentacular cells and ductal cells of Bowman's glands in the olfactory epithelium, and to the microvilli and cilia of ciliated but not secretory cells in the respiratory epithelium. No staining was detected in olfactory receptor neurons, basal cells, or two types of microvilli-bearing cells that differed from sustentacular cells. A contrasting antibody, 2H4, bound to granules of secretory respiratory cells. Developmental expression of 1F4 binding began at E17 and increased at and after E18/E19. Bulbectomy did not alter 1F4 immunoreactivity. Cell culture studies found that the 1F4 epitope was external and insensitive to trypsin treatment, and that both 1F4 and 2H4 positive cells required contact with aggregated cells for survival up to fifteen days in vitro.
The antibody 1F4 is a useful marker and potential manipulation reagent specific for sustentacular cells and their microvilli.
嗅觉上皮支持细胞可能有助于嗅觉受体神经元的存活和功能，但用于标记和操控这类细胞的试剂很少。
以全培养大鼠嗅觉黏膜细胞作为抗原刺激物，制备新型鼻细胞特异性单克隆抗体。通过大鼠组织和新生大鼠嗅觉细胞培养物的光镜免疫染色，以及成体组织冷冻置换、包埋后染色的电镜观察，对这些抗体进行表征。
一种名为1F4的IgMκ单克隆抗体在组织切片中选择性标记大鼠嗅觉上皮和呼吸上皮的顶端表面，在嗅觉细胞培养物中标记看起来像是支持细胞的细胞。利用电子显微镜观察，1F4选择性地结合嗅觉上皮中支持细胞的微绒毛和鲍曼腺的导管细胞，以及呼吸上皮中纤毛细胞而非分泌细胞的微绒毛和纤毛。在嗅觉受体神经元、基底细胞或与支持细胞不同的两种带有微绒毛的细胞中未检测到染色。一种对比抗体2H4与分泌性呼吸细胞的颗粒结合。1F4结合的发育表达在胚胎第17天开始，在胚胎第18/19天及之后增加。嗅球切除不改变1F4免疫反应性。细胞培养研究发现，1F4表位位于外部且对胰蛋白酶处理不敏感，并且1F4和2H4阳性细胞在体外存活长达15天均需要与聚集细胞接触。
抗体1F4是一种对支持细胞及其微绒毛具有特异性的有用标记物和潜在操控试剂。
