Cytochemical and Immunocytological Study on the Cellulase Activity in the Stomach of the Land Snail Nesiohelix samarangae
Kye-Heon Jeong and Yong-Seok Lee
Department of Life Sciences, College of Natural Sciences, Soonchunhyang University
In order to observe the anticellulolytic localization in the stomach tissue of a Korean land snail, Nesiohelix samarangae, a cytochemical method and a immunogold labeling method were applied. For the cytochemical study on the cellulase activity, Benedict reaction method was applied. And for the immunocytochemical study, the rabbit serum immunoglobulins (IgG) was obtained from the rabbits injected with cellulase which was extracted from body fluid of the snail. The stomach tissues of Nesiohelix samarangae were fixed with 4% paraformaldehyde and embedded in Lowicryl K4M at -40℃ under UV light (360 nm). The thin sections were loaded on the nickel grids and stained with the serum IgG and protein A-gold complex (particle size: 20 nm). Observations were undertaken with transmission electron microscope (Jeol, JEM-1010). The epithelium of the stomach was consisted of three types of cells. In the cytochemical study, the reaction products were found along the periphery of the vacuoles derived from the Bebedict reaction. In the immunocytochemical study, the protein-A gold particles were selectively labeled in Type 1 and Type 3 cells. The particular regions which were labeled with the protein-A gold particles were on the membranes of rER, in the surrounding cytoplasm of the rER and secretory granules, and in the apical cytoplasm of the cells. On the material being secreted from the apical cytoplasm was also labeled with the immunogold particles. The all results obtained throughout present study suggest that the stomach epithelium of the snail, Nesiohelix samarangae secrete cellulase as one of digestive enzymes.
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