Grote M, Vrtala S, Valenta R
Institute of Medical Physics and Biophysics, Münster University, Germany.
J Histochem Cytochem. 1993 May;41(5):745-50. doi: 10.1177/41.5.8468456.
Dry and rehydrated birch pollen grains were anhydrously fixed and double immunogold-labeled for the presence of two allergens, Bet v I major allergen (17 KD) and profilin (14 KD). In dry pollen grains, both allergens are found exclusively inside the cytoplasm. In pollen grains rehydrated for 1 min, the cytoplasm is partially devoid of the two allergens, whereas the pollen wall and the germination aperture are specifically labeled. Pollen grains rehydrated for 5 min are largely free of the two allergens. In immunoblot experiments, both allergens could be detected in the aqueous supernatants of rehydrated pollen samples within 5 min. The results obtained by both methods show the high solubility of both proteins. This makes them readily available to the immune system and characterizes them as potent allergens. Moreover, the solubilization of profilin might indicate a dissociation of the profilin-actin complex at the very first stage of pollen germination, which could favor formation of the cytoskeleton and pollen tube growth.
干燥及复水后的桦树花粉粒经无水固定，并针对两种过敏原——主要过敏原Bet v I（17千道尔顿）和肌动蛋白结合蛋白（14千道尔顿）进行双重免疫金标记。在干燥的花粉粒中，两种过敏原仅存在于细胞质内。在复水1分钟的花粉粒中，细胞质部分缺乏这两种过敏原，而花粉壁和萌发孔则被特异性标记。复水5分钟的花粉粒基本不含这两种过敏原。在免疫印迹实验中，复水花粉样品的水性上清液在5分钟内即可检测到这两种过敏原。两种方法所得结果均显示这两种蛋白质具有高溶解性。这使得它们易于被免疫系统识别，并将它们表征为强效过敏原。此外，肌动蛋白结合蛋白的溶解可能表明在花粉萌发的最初阶段肌动蛋白结合蛋白 - 肌动蛋白复合物发生了解离，这可能有利于细胞骨架的形成和花粉管的生长。
