Can hypoosmotic shock and calcium influx lead to translocation of aquaporin-1 in shrimp muscle cells?

The physiological variations during the crustacean molting cycle have intrigued researchers for many years. Maintaining osmotic homeostasis in the face of hemolymph dilution and dealing with dynamic intracellular and extracellular calcium fluctuations are challenges these animals continuously confront. It has recently been shown that water channels present in the cell membrane (aquaporins) are essential for water uptake during premolt and postmolt. This study aims to investigate whether hypoosmotic shock and intracellular and extracellular calcium variations can lead to translocation of Aquaporin 1 (AQP-1) from the intracellular region to the plasma membrane during premolt and postmolt, thus allowing increased water flow in these stages. For this, we investigate in vitro the rapid change of AQP-1 positions in the abdominal muscle cells in the freshwater shrimp, Palaemon argentinus. Using cell volume analysis and immunohistochemistry, we show that hypoosmotic conditions and an elevation of the intracellular and extracellular calcium concentrations are concurrent with the translocation of AQP-1 to the plasma membrane. These results indicate that calcium flux and hypoosmotic shock may be regulators of AQP 1 in the translocation process.

Can short-term exposure to copper and atrazine be cytotoxic to microalgae?

Aquatic environments can be easily contaminated due to anthropogenic activities that may affect local biota. Microalgae are abundant and have an important role on the food chain. Consequently, they stand out as promising models for studies of contaminants. This study investigated the cytotoxic effects of atrazine and copper (separate and mixture) exposure in microalgae Desmodesmus communis, as well as its cellular defense due to ABC (ATP-binding cassette) proteins activity against the xenobiotics. We analyzed two different ABC proteins activity pathways: P-gp, which is responsible for nonspecific substance efflux, and MRP that is associated with metals efflux. It was observed that the microalgae exposure to atrazine (90 nM) and copper (141 nM) has been considered cytotoxic. When contaminants were mixed, only the combination of both highest concentrations tested was cytotoxic. The P-gp blocker, verapamil, demonstrated that the contaminants tested caused proteins inhibition. However, the MK-571 (MRP blocker) did not block pump activity. There was an inverse relationship between ABC protein activity and cytotoxicity; non-cytotoxic conditions suggest increased activity of microalgae defense proteins.

Toxicity induced by glyphosate and glyphosate-based herbicides in the zebrafish hepatocyte cell line (ZF-L).

Glyphosate is the active component of many commonly used herbicides; it can reach bodies of water through irrigated rice plantations. In the present study, we evaluated the effect of glyphosate and Roundup® (a glyphosate-based herbicide) in established culture of the zebrafish hepatocyte cell line ZF-L after 24 and 48 h of exposure to concentrations of 650 and 3250 µg/L. We observed a reduction in metabolic activity and lysosomal integrity, and an increase in cell number after 24 h of Roundup® exposure at the highest concentration. An increase in active mitochondria and apoptotic cells was observed following 24 h exposure to glyphosate and Roundup®, while only exposure to Roundup® induced an increase in necrotic cells. Rhodamine B accumulation decreased after 48 h exposure to 650 µg/L of Roundup®; this reduction is indicative of increased activity of ABC pumps. Overall, the present findings highlighted the hazard of glyphosate exposure not only in the commercial formulation but also glyphosate alone, since both can induce damage in the ZF-L cell line primarily through the induction of apoptosis.

Glyphosate Adversely Affects Danio rerio Males: Acetylcholinesterase Modulation and Oxidative Stress.

It has been demonstrated that glyphosate-based herbicides are toxic to animals. In the present study, reactive oxygen species (ROS) generation, antioxidant capacity against peroxyl radicals (ACAP), and lipid peroxidation (LPO), as well as the activity and expression of the acetylcholinesterase (AChE) enzyme, were evaluated in Danio rerio males exposed to 5 or 10 mg/L of glyphosate for 24 and 96 h. An increase in ACAP in gills after 24 h was observed in the animals exposed to 5 mg/L of glyphosate. A decrease in LPO was observed in brain tissue of animals exposed to 10 mg/L after 24 h, while an increase was observed in muscle after 96 h. No significant alterations were observed in ROS generation. AChE activity was not altered in muscles or brains of animals exposed to either glyphosate concentration for 24 or 96 h. However, gene expression of this enzyme in the brain was reduced after 24 h and was enhanced in both brain and muscle tissues after 96 h. Thus, contrary to previous findings that had attributed the imbalance in the oxidative state of animals exposed to glyphosate-based herbicides to surfactants and other inert compounds, the present study demonstrated that glyphosate per se promotes this same effect in zebrafish males. Although glyphosate concentrations did not alter AChE activity, this study demonstrated for the first time that this molecule affects ache expression in male zebrafish D. rerio.

Effect of glyphosate on the sperm quality of zebrafish Danio rerio.

Glyphosate is a systemic, non-selective herbicide widely used in agriculture worldwide. It acts as an inhibitor of the enzyme 5-enolpyruvylshikimate-3-phosphate synthase by interrupting the synthesis of essential aromatic amino acids. This pathway is not present in animals, although some studies have shown that the herbicide glyphosate can affect fish reproduction. In this study, the effect of glyphosate on sperm quality of the fish Danio rerio was investigated after 24 and 96 h of exposure at concentrations of 5mg/L and 10mg/L. The spermatic cell concentration, sperm motility and motility period were measured employing conventional microscopy. The mitochondrial functionality, membrane integrity and DNA integrity were measured by fluorescence microscopy using specific probes. No significant differences in sperm concentration were observed; however, sperm motility and the motility period were reduced after exposure to both glyphosate concentrations during both exposure periods. The mitochondrial functionality and membrane and DNA integrity were also reduced at the highest concentration during both exposure periods. The results showed that glyphosate can induce harmful effects on reproductive parameters in D. rerio and that this change would reduce the fertility rate of these animals.

Effects of glyphosate on cholinesterase activity of the mussel Perna perna and the fish Danio rerio and Jenynsia multidentata: in vitro studies.

Although the herbicide glyphosate [N-(phosphonomethyl)glycine] is not classified as an acethylcholinesterase inhibitor, some studies have reported reduction in the acethylcolinesterase activity after in vivo exposure to both its pure form and its commercial formulations. Considering this controversy, the objective of the present study was to investigate, in vitro, the effects of glyphosate exposure on cholinesterase activity of the brown mussel Perna perna and of two fish species: zebrafish Danio rerio and onesided livebearer Jenynsia multidentata. For this purpose, samples of different tissues (brain and muscle for fish; gills and muscle for mussel) were homogenized and pre-incubated with different glyphosate concentrations before cholinesterase activity determination. Results demonstrated that cholinesterase from different fractions of all species tested was inhibited by glyphosate. The concentrations of glyphosate that inhibits 50% of cholinesterase activity (IC50) ranged from 0.62 mM for P. perna muscle to 8.43 mM for J. multidentata brain. According to this, cholinesterase from mussel seems to be more sensitive to glyphosate exposure than those from the fish D. rerio and J. multidentata.