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Immunocytochemical localization of Cry j 1, the major allergen of Cryptomeria japonica (Taxodiaceae) in Cupressus arizonica and Cupressus sempervirens (Cupressaceae) pollen grains

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Abstract

Immunocytochemical localization of cross-reactive antigens to Cry j 1, the major allergen of Cryptomeria japonica, in the pollen grains of Cupressus arizonica and Cupressus sempervirens, was conducted using transmission electron microscopy. Mature and activated pollen grains were fixed for immunocytochemistry using conventional and freezing protocols. Sections containing the samples were incubated with anti-Cry j 1 monoclonal antibody (mAb) (KW-S91). Cross-reactive antigens to Cry j 1 were detected in the mature pollen grains of Cupressus arizonica and Cupressus sempervirens, and abundant gold particles were seen in the orbicules and wall, and in the Golgi, nucleus, and inclusions in reserve materials. After 5 min of activation, labelling noticeably decreased. From 15 min to 48 h, the cytoplasm exhibits a new labelling pattern, with gold markers being associated with protein storage vacuoles. The decrease in cross-reactive antigens during the first 5 min of activation could be due to a rapid release of proteins recognized by Cry j 1 mAb during pollen attachment in the pollination droplet. Our results show that the content of allergenic proteins is unstable, displaying variation relative to the progress of germination in Cupressus sempervirens and Cupressus arizonica pollen grains. The ability to do so may be viewed as an adaptive strategy of Cupressaceae pollen grains to maximize their biosynthetic efficiency.

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Fig. 1a–f.
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Acknowledgements

The authors would like to thank Dr. T. Nagoya of Kowa Research Institute, Kowa Co., Ltd., Tsukuba, Japan for kindly supplying anti-Cry j 1 mAb (KW-S91) and are grateful to the Scientific Technical Services at the University of Barcelona for their careful preparation of the sections for TEM. This study was supported by Grant BOS2000-0563-C02, Ministry of Science and Technology, Spain.

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Authors and Affiliations

  1. Department of Botany, Faculty of Pharmacy, University of Barcelona, Av. Diagonal s/n, 08028 , Barcelona, Spain

    María Suárez-Cervera, Ana Vega-Maray & Juan A. Seoane-Camba

  2. Yamagata Prefecture, Institute of Public Health, 990-0031, Yamagata , Japan

    Yuichi Takahashi

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  1. María Suárez-Cervera
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  2. Yuichi Takahashi
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Correspondence to María Suárez-Cervera.

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Suárez-Cervera, M., Takahashi, Y., Vega-Maray, A. et al. Immunocytochemical localization of Cry j 1, the major allergen of Cryptomeria japonica (Taxodiaceae) in Cupressus arizonica and Cupressus sempervirens (Cupressaceae) pollen grains. Sex Plant Reprod 16, 9–15 (2003). https://doi.org/10.1007/s00497-003-0164-x

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