CELLULAR LOCALIZATION OF MARINE-DERIVED EXCITATORY AMINO ACIDS IN THEIR PRODUCING ORGANISMS

点击翻译按钮获取中文摘要

Cellular localizations of two representative marine excitatory ammo acids : dysiherbaine (DH) and kainic acid (KA) were investigated using immunohistochemical methods. Selective polyclonal antibodies to each amino acid were raised by coupling each amino acid to carrier protein such as KLH. Using successfully prepared antibodies, section of Micronesian sponge Dysidea herbacea and red algae Digenea simplex, producing organism of DH and kainic acid, respectively, were separately stained then visualized with protein A-gold. Microscopic observations indicated that DH was localized exclusively in the spherical cell present hi the mesohyl of the sponge.We also showed a presence of putative DH-binding protein of 111kD using DH-conjugated affinity chromatography and SDS-PAGE. We preliminary observed by RT-PCR using GluR-targeted primers that there might be a GluR-like molecule in the sponge.KA was mainly located in the cells of heir-like structure of the algae which grows from the main stem of the algae. Only very surface of the main stem was stained. The surface of the stem which likely be transforming into the hear-like structure was stained densely. The hear-like structure composed of three cell layers : outer small cells, middle layer cells and the central cells. Numbers of chloroplasts were present in the outer small cells. KA-stain was observed all these cells. A small (1 to 2 μm) granular body observed in the surrounding cells was stained somewhat stronger then other cellular structures.