Metallothionein determination can be applied to learn about aquatic metal pollution and oxidative stress detoxification mechanisms through Problem-based Learning.

A simple, repeatable and inexpensive laboratory practice applied to teach and discuss aquatic metal pollution and oxidative stress detoxification mechanisms through biomarker analyses, as well as important ecotoxicology concepts, is presented herein. It has been implemented in a university in Brazil to both undergraduate and Master's and PhD students, indicating usefulness to all these levels. Students learned to detect metallothionein and reduced glutathione concentrations in biological samples and investigate several variables of interest in biomonitoring assessments. In addition, statistical correlations were used to indicate the potential dual role played by MT in aquatic organisms, allowing for biological inferences regarding both aquatic metal pollution and oxidative stress detoxification mechanisms and maturing of ecotoxicological and biomonitoring concepts discussed and presented both theoretically and integrated to the laboratory findings.

Identification and tissue-specific distribution of sulfated glycosaminoglycans in the blood-sucking bug Rhodnius prolixus (Linnaeus).

We have previously characterized heparan sulfate (HS) as the major ovarian sulfated glycosaminoglycan (GAG) in females of Rhodnius prolixus, while chondroitin sulfate (CS) was the minor component. Using histochemical procedures we found that GAGs were concentrated in the ovarian tissue but not found inside the oocytes. Here, we extend our initial observations of GAG expression in R. prolixus by characterizing these molecules in other organs: the fat body, intestinal tract, and the reproductive tracts. Only HS and CS were found in the three organs analyzed, however CS was the major GAG species in these tissues. We also determined the compartmental distribution of GAGs in these organs by histochemical analysis using 1,9-dimethylmethylene blue, and evaluated the specific distribution of CS within both male and female reproductive tracts by immunohistochemistry using an anti-CS antibody. We also determined the GAG composition in eggs at days 0 and 6 of embryonic development. Only HS and CS were found in eggs at day 6, while no sulfated GAGs were detected at day 0. Our results demonstrate that HS and CS are the only sulfated GAG species expressed in the fat body and in the intestinal and reproductive tracts of Rhodnius male and female adults. Both sulfated GAGs were also identified in Rhodnius embryos. Altogether, these results show no qualitative differences in the sulfated GAG composition regarding tissue-specific or development-specific distribution.