Effects of juglone (5-hydroxy-1,4-naphthoquinone) on midgut morphology and glutathione status in Saturniid moth larvae.

Actias luna and Callosamia promethea larvae were fed birch foliage supplemented with juglone (5-hydroxy-1,4-naphthoquinone) to determine whether juglone causes oxidative stress in midguts of these species. Juglone is a substituent of walnut foliage. A. luna, but not C. promethea, thrives on walnut foliage, as well as birch foliage supplemented with juglone. After 2 and 3 days on juglone-containing diets, midgut samples from these animals were compared histologically and were analyzed for GSH and GSSG content. C. promethea, but not A. luna, midguts revealed partial loss of epithelial structure. In contrast, GSH and GSSG did not change significantly in either species. In a separate experiment, live midgut explants from each species were cultured for 4 h in 0, 0.05, and 0.25% juglone. In juglone-treated explants, GSSG increased 2.1 and 5.6-fold, respectively, for A. luna, and 1.6 and 2.7-fold, respectively, for C. promethea. There was also a small dose-dependent decrease in GSH in C. promethea, but not A. luna. Although histology indicates that the midgut is a target of juglone toxicity in C. promethea, GSH analyses from either species do not support the expectation that changes in GSH/GSSG explain differences in susceptibility to juglone toxicity.

Schistosoma japonicum GSH S-transferase Sj26 is not the molecular target of praziquantel action.

It has been suggested that Sj26, a Schistosoma japonicum GSH S-transferase, is the molecular target of the antischistosomal drug praziquantel (McTigue et al., 1995, J. Mol. Biol. 246, 21-27). We tested this hypothesis by asking two questions: (1) does praziquantel inhibit Sj26 activity with a variety of model substrates; and (2) does praziquantel prevent the binding to Sj26 of physiologically relevant nonsubstrate ligands? High concentrations of praziquantel (up to 500 microM) did not inhibit Sj26 activity using the model substrates 1-chloro-2,4-dinitrobenzene, 3,4-dichloronitrobenzene, or ethacrynic acid. Sj26 had no measurable activity with two higher molecular weight GSH S-transferase substrates: 5-androsten-3,17-dione and sulfobromophthalein. We also assessed the ability of praziquantel to prevent the inhibition of Sj26 by a series of S-alkyl-GSH conjugates. The half-maximal inhibitory concentrations of S-hexyl-GSH, S-octyl-GSH, and S-decyl-GSH (10, 10, and 5 microM, respectively) for Sj26 were not affected by up to 500 microM praziquantel. This suggests that praziquantel does not compete with GSH for Sj26 binding. In order to determine if praziquantel disrupts binding of nonsubstrate ligands to Sj26, we tested praziquantel for its ability to prevent the inhibition of Sj26 by both bilirubin and hematin. Praziquantel (100 or 500 microM) did not alter inhibition of Sj26 by 3 microM bilirubin, but partially protected Sj26 against inhibition by hematin (0.1 to 2.0 microM). Interestingly, in a similar reaction, 100 microM S-methyl-GSH protected Sj26 from inhibition equally as well as praziquantel. Bovine serum albumin (5 microM) completely protected against inhibition by 1 microM hematin. These results indicate that although praziquantel partially protects Sj26 from hematin inhibition, this protection is neither specific to praziquantel nor physiologically relevant. Our results do not support the hypothesis that the mechanism of praziquantel action involves competitive inhibition of Sj26 catalytic activity or blocking binding of nonsubstrate ligands. We can, therefore, find no evidence that Sj26 is the molecular target of the antischistosomal activity of praziquantel.

Differential toxicity of juglone (5-hydroxy-1,4-naphthoquinone) and related naphthoquinones to saturniid moths.

The preferred hosts of the saturniid mothActias luna include members of the Juglandaceae, whose foliage contain the toxin juglone (5-hydroxy-1,4-naphthoquinone). The performance ofActias luna andCallosamia promethea was compared when fourth-instar larvae of each were fed birch foliage, a mutually acceptable food plant, or birth supplemented with 0.05% (w/w) juglone.A. luna fed juglone exhibited no changes in developmental time or mortality compared to a diet without juglone. In contrast, juglone-supplemented diets, when fed toC. promethea, caused negative growth rate, and a 3.6-fold decrease in consumption rate. The performance ofA. luna also was compared on birch and walnut; larvae developed and grew more rapidly on an all-walnut vs. an all-birch diet. To examine the effect of 1,4-naphthoquinone structure onA. luna survival, first instars were fed on birch supplemented with varying concentrations of juglone (J), menadione (M), plumbagin (P), or lawsone (L). In diets supplemented at 0.05% (w/w), none of the compounds produced effects significantly different from controls. In diets supplemented at 0.5% (w/w), the treatments produced significant toxic effects in the order P>M=L>J for mortality, and P>L>M=J for increased developmental time. Late-instarA. luna are clearly resistant to juglone compared toC. promethea, and early-instarA. luna are resistant to several related 1,4-naphthoquinones. These results suggest a chemical basis for host choice among saturniids. In addition, the luna-walnut system may be a valuable model for studying quinone detoxication.