Superoxide production in insect hemolymph plasma (iv)

Seventh JADCI abstracts

AND ITS DEFENSIVE IN THE JAPANESE

III

FUNCTION

FRESHWATER CRAB (Potamon dehauni) Michiko Kimura I-IEMOLY MPH. and Hatuhisa Wago. Department of Saitama Medical Me&al Technology, School Junior College, Moroyamq IrumaWe have gun, Saitama 350-04, Jqan. some sugars such as Dreported that mannose, N-accthyl-D-glucosamine, Nacetyl-D-galactosamine and I.-fucose are cytoplasmic granules of present in granulocytes and semigranulocytes in Potmnon dehaani. Degranulated hemocytes absorbed animal erythrocytes more strongly in the presence of hemolymph. From these results, it appeared that hemolymph absorption is due to the recognition of both granule sugar and erythrocyte by plasma lectin. Thus, this study was carried out to reexamine the presence of lectin and its physicochemical properties using plasma obtained from crab claws. Results showed that: 1) lectin agglutinating human type-0 and sheep erythrocytes occurred in plasma; 2) N-acetyl-D-glucosamine or N-acetyl-Dgalactosamine inhibited lectin activity; 3) lectin activity disappeared by heat 100°C . Moreover, since treatment of EDTA treatment strongly suppressed lectin activity, lectin was considered to be a C-type lectin requiring Ca2+. These results suggested that lectin is involved in adhesive reactions such as hemarkorption and this occurs by recognizing some sugars in the granules released after degranulation. Moreover, we also report the sensitivity of lectin to protease, the inhibitory effect of sugars on hemadsorption by crab hemocytes and the analysis of lectin by SDS-PAGE.

Previously, produce superoxide anion. superoxi& has been reported to generate in Pseutietia separata hemolymph plasma in vitro. Hem, superoxide production was also detected in five other 1epiQpteran species, Mamestnl that is, Spodoptem lituq brukcae, Agrius convolvuli, Bombyx mori, and Pietis nqae crucivonz Superoxide generative system in plasma consists of at least two components, low (LMF) and high (HMF) molecular weight factors. HMF was purified from B. mori larval plasma by dialysis (MW-300,000 cut off), gel-filtration (Sephacryl S300HR) aud KEk density gradient ultracentrifngation. Specific gravity of the HMF was 1.12, which coincided with the density of B. mori lipophorin. Two subunits of the lipophorin were detected by SDSPAGE analysis of the fraction (d=1.12). It is of interest that whether a superoxide production in plasma and a participation of plasma lipoproteins in that process are widely detected in other invertebrates and vertebrates.

A4. INVOLVEMENT OF GRANULOCYTES AND PLASMATOCYTES FOR PHAGOCYTIC REACTION IN LARVAE OF TWO LEPIDOPTERAN SPECIES. Shinya Yokoo, Peter Giitz*, Hiromi Nishimura and Sumio Tojo. Faculty of Agriculture, Saga University, Saga 840, Jqan and *Free Univssity of Berlin, D14195, Berlin, Gennany. Differential cell fractionation (D. F.) and nylon wool column (N.W.) methods, both utilizing different ion calcium requirements among immunocytes for adhesion to non-self, were adopted to prepare immunocyte monolayers from Agrotis segetum and Galleria mellonella larvae. Monolayers prepared by the D.F. and A3. SUPEROXIDE PRODUCTION N.W. methods showed high purities of IN INSECT HEMOLYMPH plasmatocytes granular (PL) and PLASMA (IV). Toru Arakawa. plasmatocytes (GPL). Granulocytes separated National Institute of Sericultural and by the D.F. method in vitro showed high Entomological Science, Tsukuba, lbamki phagocytic activities against lndian ink 305, Jqoan. Superoxide production by particles and FlTC-labeled silica beads, while mammalian polymorphonuclear hemocytes is PL and GPL prepared by both methods known to be essential for killing bacteria. actively engulfed only FITC-labeled silica Hemocytes of invertebrate species also beark. Essentially similar results were