Pablo Elías Martínez

Effects of prenatal and postnatal exposure to a low dose of bisphenol A on behavior and memory in rats.

Endocrine disruptors (EDs) are increasingly common chemicals in the environment. Bisphenol A (BPA), used to manufacture polycarbonate plastics, is an ED recognized for its estrogenic, anti-estrogenic, and anti-androgenic effects. Behavior is considered a vital characteristic for an animals life cycle. This study evaluated the effect of exposure to low doses of BPA during pregnancy and/or lactation on several aspects of rat behavior, including memory, locomotion, and the exploratory instinct. Pups at 16 weeks of age (females and males) were divided into groups according to the mothers exposure to BPA (40μg/kg/day): CON (vehicle only); PRE (during pregnancy); LAC (during lactation); PRE-LAC (during both pregnancy and lactation). In the PRE-LAC group, exposure to BPA impaired both short-term (STM) and long-term memory (LTM) in inhibitory avoidance and the object recognition task, and also affected locomotor activity and spatial memory. Some sex-specific behavioral characteristics disappeared in the LAC group. Sex-specific memory and behavior impairment were caused by BPA exposure during brain organogenesis and differentiation.

Lactational transfer of PCBs and chlorinated pesticides in pups of southern elephant seals (Mirounga leonina) from Antarctica

Seven pairs of southern elephant seals (Mirounga leonina) pups and their dams were sampled during the late weaning season among a breeding population of seals on Elephant Island in Antarctica. The blubber of the pups and the milk and blubber of their dams were analyzed for lipid-normalized concentrations of PCBs and organochlorines compounds in order to evaluate the lactational transfer of these contaminants. The lipid-normalized concentrations in these tissues were in the ppb range (i.e., ngg(-1) lipid). The levels of contaminants in southern elephant seals were low in comparison with residues that have been reported in pinnipeds from the northern hemisphere. The relative tissue concentrations of the analytes measured followed the pattern: SigmaDDT>mirex>SigmaPCB>Sigmachlordane>HCB>heptachlor epoxide>dieldrin>methoxychlor>SigmaHCH>other organochlorines. The very high DDE/SigmaDDT ratio (0.91) in the blubber of dams and pups was an indicative of long-term, extremely distant pollution. On the other hand, the relatively high levels of some other organochlorine pesticides (e.g. mirex, heptachlor epoxide, dieldrin, methoxychor) may reflect the continued use of these insecticides in developing countries located in the southern hemisphere. For most of the analytes measured, the lipid-normalized concentrations were lower in pup blubber and in the milk than in the maternal blubber. Lactational transfer rates were dependent on the logK(ow) (octanol/water partition coefficient) values of the analytes measured, less lipophilic compounds being more readily transferred to the pups by the lactational route.

Effects of increasing temperature on antioxidant defense system and oxidative stress parameters in the Antarctic fish Notothenia coriiceps and Notothenia rossii

Antioxidant defense system (ADS) and oxidative stress parameters were evaluated in the Antarctic fish Notothenia rossii and N. coriiceps exposed to increasing temperature. Acclimated fish were kept at 0°C or exposed to 4°C for 1day (N. rossii) or to 2 and 4°C for 1 and 6 days (N. coriiceps). Measurements were assessed in brain, gills, liver, white muscle and erythrocytes. Parameters analyzed included antioxidant capacity against peroxyl radicals (ACAP); reduced glutathione (GSH) and metallothionein-like proteins (MTLP) concentration; superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and glutamate-cysteine ligase (GCL) activity; lipid peroxidation (LPO) level and protein carbonyl (PC) concentration. Increased liver GST activity was observed in N. rossii exposed to 4°C for 1day. Increased muscle GPx activity was observed in N. coriiceps after exposure to 2°C for 1day. Reduced gill GPx activity and increased liver SOD activity were observed after exposure to 4°C for 1day. In N. coriiceps, increased gill GCL activity and reduced gill GPx activity, as well as reduced liver MTLP were observed after exposure to 2°C for 6 days. Reduced brain SOD activity and increased brain LPO; reduced gill ACAP, GSH concentration and GPx activity, as well as increased gill GCL activity; reduced liver ACAP, MTLP, SOD activity, GST activity and increased liver and erythrocytes LPO were observed after exposure to 4°C for 6 days. These findings indicate that ADS is more responsive to short-term increasing temperature in the sluggish N. coriiceps than in the active N. rossii. However, responses of N. coriiceps to long-term increasing temperature were transient and did not prevent tissue oxidative damage. Considering the predicted increase in temperature in the Southern Ocean over the next decades, our findings suggest that Antarctic fishes are sensitive to ocean warming, displaying tissue oxidative damage associated with the thermal stress.

Sperm impairments in adult vesper mice (Calomys laucha) caused by in utero exposure to bisphenol A

This study aimed to evaluate the effects of in utero administration of bisphenol A (BPA) on semen parameters of vesper mice. Sixty female Calomys laucha were divided into six groups and received by gavage during gestation the following substances: Water (negative control), Olive Oil (vehicle control), Diethylstilbestrol (DES – positive control – 6.5 μg kg−1 bw) and BPA (40, 80 and 200 μg kg−1 bw). Male offspring were euthanised at 70 days of age, and sperm parameters were analysed. BPA reduced normal sperm morphology (water = 96.1 ± 0.65; BPA200 = 96.8 ± 2.3%), sperm membrane integrity (water = 88.8 ± 1,65; BPA200 = 70.6 ± 4,15%), sperm motility (water = 87.5 ± 1.71; BPA200 = 51.3 ±9.9%) and in vitro penetration rates (water = 55.0 ± 7.14; BPA200 = 7.47 ±2.96%), but it did not affect body weight, anogenital distance, sperm DNA integrity and acrosome integrity. In conclusion, in utero exposure to BPA caused a reduction in sperm parameters of adult C. laucha. Natural mating studies should be conducted to verify the effects of BPA on fertility of the animals.

The effects of aging on leukocyte glucocorticoid receptor concentration and response to dexamethasone in dogs

Glucocorticoid receptor concentration (GR) was determined in mononuclear (MNS) and polymorphonuclear (PMNS) cells isolated from 8 aged and 9 young male dogs. In addition, leukocyte responses to dexamethasone (0.1 mg kg(-1) i.v.) and plasma glucose concentration were also measured. The GR in MNS and PMNS was higher (p<0.05) in young dogs (6.64 +/- 0.57 and 7.04 +/- 0.29 fmolesx10(6) cells, respectively, versus 4.40 +/- 0.24 and 5.06 +/- 0.33 fmolesx10(6) cells, respectively, in aged dogs). The maximum increase in neutrophils (DeltaNEU) was lower (p<0.05) in aged dogs 6003.38 +/- 1398.5x10(6) versus 11168.67 +/- 1863.16x10(6) cells l(-1) in young dogs. The maximum decrease in lymphocytes (DeltaLYM) was lower (p<0.05) in aged dogs 550 +/- 56.75x10(6) cells l(-1) versus 1825.89 +/- 313.1x10(6) cells l(-1) in young dogs. In young dogs, significant (p<0.05) correlations between GR in PMNS and DeltaNEU (r=0.80) and between GR in MNS and DeltaLYM (r=0.76) were observed. In aged dogs, these correlations were not significant. The lower GR value and the lack of correlation between this parameter and its respective Delta in aged dogs suggest that changes in leukocytes responsiveness to glucocorticoids is occurring during the aging process.

Antioxidant defense system and oxidative status in Antarctic fishes: The sluggish rockcod Notothenia coriiceps versus the active marbled notothen Notothenia rossii

Adaptive responses of antioxidant defense systems (ADS) to changes in increased levels of activity are critical, especially in Antarctic fishes. The benthopelagic marbled notothen (Notothenia rossii) shows higher spontaneous activity than the benthonic and sluggish rockcod (N. coriiceps). Therefore, we hypothesize that species-related responses of ADS would occur to counteract different rates of reactive oxygen species formation in these two Antarctic fish. Here we evaluated ADS and oxidative damage in tissues (brain, gills, liver and white muscle) of the two Antarctic fish. Despite no significant differences in lipid and protein oxidative damage were observed, we actually found species- and tissue-specific differences in ADS. Gill metallothionein-like proteins (MTLP) and liver reduced glutathione (GSH) concentrations were higher in N. coriiceps than in N. rossii. Brain and gill antioxidant capacity against peroxyl radicals (ACAP); gill enzyme [glutamate-cysteine ligase (GSL), superoxide dismutase (SOD) and catalase (CAT)] activity; liver GCL and SOD activity; and white muscle CAT activity were higher in N. rossii than in N. coriiceps. Therefore, the more active fish (N. rossii) maintains higher activities of enzymes involved in superoxide ions (O2.-) detoxification and GSH production in peripheral tissues (gills, liver and white muscle). This allows the more active fish (N. rossii) to keep levels of lipid and protein oxidative damage similar to those observed in the sluggish fish (N. coriiceps). It is worth noting that the more active fish also shows a higher brain antioxidant capacity, which could involve other non-enzymatic antioxidants like vitamins C and E. In contrast, N. coriiceps shows lower consumption of non-enzymatic antioxidants in peripheral tissues than N. coriiceps. As hypothesized, our results indicate that differences in ADS profiles between fish species are likely related to their habits and metabolic rates. This would imply in different fish abilities to deal with oxidative stress associated with increasing seawater temperature.

Effects of seasonality and moult cycle on the proliferation of nerve cells and on the labelling of ecdysone receptors in an estuarine crab

Decapod crustaceans show proliferation of the nerve cells in the olfactory lobe throughout their lives. However, the regulation of this process is still poorly understood, since it may vary with endogenous and exogenous factors. The objective of the present investigation was to quantify the proliferation of nerve cells and number of nerve cells with ecdysone receptors in the clusters of the central olfactory system in Neohelice granulata, according to moult stages and in different seasons (summer and winter). Three injections of bromodeoxyuridine (BrdU) were administered to the crabs. Brains were sectioned by microtome and fixed on slides for immunohistochemistry with anti-BrdU and anti-EcR antibodies. The proliferation of nerve cells was higher in winter than in summer, probably because in winter the crabs do not breed and the premoult and postmoult periods are longer. Crabs in postmoult exhibited more BrdU-labelled cells than crabs in premoult or intermoult in winter, because of a greater number of mitoses related to an increase in body size and addition of olfactory receptor neurons. The number of EcR-labelled cells was higher in premoult than in postmoult or intermoult in winter. The proliferation of nerve cells is regulated seasonally and according to moult stages.