Maria Teresa Colomina

Effects of oral aluminum exposure on behavior and neurogenesis in a transgenic mouse model of Alzheimer's disease

The effects of a very low oral dose of Al on spatial learning and neurogenesis were evaluated in a transgenic mouse (Tg 2576) model of Alzheimer disease. At 5 months of age, wild and Tg 2576 mice received a diet supplemented with Al lactate at 0 and 1 mg/g of diet for 120 days. The experimental groups (n=7-8) were: control wild, Al-treated wild, control transgenic, and Al-treated transgenic. After 3 months of Al exposure, activity in an open-field and learning in a water maze were evaluated. At the end of the behavioral testing, in order to study cell proliferation and differentiation in the hippocampus, mice were injected with 5-bromo-2-deoxyuridine (BrdU) and sacrificed 1 or 28 days after the last BrdU injection. Tg 2576 mice were impaired in both acquisition and retention of the water maze task, showing higher amounts of beta-amyloid fragments in brain. Aluminum exposure impaired learning and memory in wild mice and increased the total number of proliferating cells in the dentate gyrus of hippocampus. The low Al doses here experimented suggest that this element might impair cognition in the general population at doses comparable to current levels of human exposure. Although these doses are not enough to interact with the amyloidogenic pathway, an increase in cell proliferation can indicate a reactive response of the brain to Al insult. Further investigations should be performed to corroborate the effects observed at very low doses of Al and to study the potential effects derived from a longer exposure period.

Reproductive toxicity evaluation of vanadium in male mice

The reproductive toxicity of vanadium was studied in mice. Male Swiss mice were exposed to sodium metavanadate at doses of 0, 20, 40, 60, and 80 mg/kg per day given in the drinking water for 64 days. To evaluate the fertility of the vanadium-treated animals, males were mated with untreated females for 4 days. A significant decrease in the pregnancy rate was observed at 60 and 80 mg/kg per day of sodium. metavanadate. However, metavanadate did not reduce fertility in male mice at 20 and 40 mg/kg per day. Reproductive toxicity was measured by sperm count, sperm motility, organ weights, and histologic evaluation of the testes. Decreased body and epididymis weight was only observed in the 80 mg/kg per day group, while testicular weights were not altered by the treatment with all doses used. Sperm count was significantly decreased at 40, 60, and 80 mg/kg per day, but the sperm motility was unaffected. Histopathological examination revealed that the testes were normal and that the epididymis of treated male mice contained normal appearing sperm. The no observed adverse effect level (NOAEL) was 40 mg/kg per day. Consequently, vanadium would not cause any adverse effect on fertility or testicular function in male mice at the concentrations usually ingested by humans through the diet and drinking water.

Influence of Maternal Stress on the Effects of Prenatal Exposure to Methylmercury and Arsenic on Postnatal Development and Behavior in Mice: A Preliminary Evaluation

The present study combined maternal restraint stress with exposure to 2 environmental toxic elements, mercury and arsenic, given to mice concurrently with the restraint period (1000-1200 h, gestational days 15-18). Two groups of animals were given, by gavage, methylmercury chloride (MMC) (2 mg/kg/day), and 2 additional groups received sodium arsenite (10 mg/kg/day) on days 15-18 of gestation. Immediately after MMC or arsenite exposure, 1 group of MMC-treated mice and 1 group of arsenite-treated animals were restrained for 2 h/day. Control groups included restrained and unrestrained pregnant mice nonexposed to MMC or arsenite. All animals were allowed to deliver and wean their offspring. Pups were evaluated for physical development, as well as for behavioral effects. Except for a significant decrease in pivoting on postnatal day 9 in the group exposed to arsenite plus restraint, no other MMC- or arsenite-induced behavioral changes were noted in unrestrained or restrained groups. Although a significant delay in pinna detachment and in eye opening was observed in pups of the group exposed to arsenite and restraint, the development landmarks were not affected by restraint in the MMC-treated animals. Although maternal stress reduced body weight gain in the dams exposed to MMC plus restraint, a significant interaction between maternal stress and MMC could not be established for developmental toxicity. These preliminary results, combined with those of previous investigations, show that stress can significantly exacerbate the adverse effects of environmental toxic elements.

Effects of maternal stress on methylmercury-induced developmental toxicity in mice

The developmental toxicity of combined exposure to maternal restraint stress and methylmercury chloride (MMC) was assessed in Swiss mice. On day 10 of gestation, four groups of plug-positive female mice were treated (p.o.) with a single dose of 12.5 or 25 mg MMC/kg. Immediately after MMC exposure, two of those groups were subjected to restraint for 14 hr. Control groups included restrained and unrestrained pregnant mice nonexposed to MMC. Combined exposure to 25 mg MMC/kg and restraint enhanced MMC-induced maternal toxicity, which included deaths and decreased body weight gain and food consumption. The number of nonviable implants was also increased significantly following concurrent exposure to MMC (25 mg/kg) and restraint, with the percentage of postimplantation loss increased from 64% (MMC alone) to 100% (MMC plus restraint). However, the types and incidence of internal and skeletal anomalies observed after administration of 12.5 mg MMC/kg were not increased by maternal restraint. These results suggest that maternal stress would enhance the MMC-induced maternal and embryo/fetal toxicity at doses of MMC that are highly toxic to the dams, whereas at doses that are less acutely toxic the role of maternal stress would not be significant.

Organophosphate pesticide exposure and neurodegeneration.

Organophosphate pesticides (OPs) are used extensively throughout the world. The main sources of contamination for humans are dietary ingestion and occupational exposures. The major concerns related to OP exposure are delayed effects following high level exposures as well as the impact of low level exposures during the lifespan which are suggested to be a risk factor for nervous system chronic diseases. Both high and low level exposures may have a particularly high impact in population subgroups such as aged or genetically vulnerable populations. Apart from the principle action of OPs which involves inhibition of the acetylcholinesterase (AChE) enzyme, several molecular targets, such as hormones; neurotransmitters; neurotrophic factors; enzymes related to the metabolism of beta amyloid protein as well as inflammatory changes have been identified for OP compounds. Here we review the main neurological and/or cognitive deficits described and the experimental and epidemiological relationships found between pesticide exposure and Alzheimers, Parkinsons, and Amyotrophic Lateral Sclerosis (ALS) diseases. This report also focuses on possible individual differences making groups resilient or vulnerable to these toxicants. A critical discussion of the evidence obtained from experimental models and possible sources of bias in epidemiological studies is included. In particular this review aims to discuss common targets and pathways identified which may underlie the functional deficits associated with both pesticide exposure and neurodegeneration.

Assessing anxiety in C57BL/6J mice: a pharmacological characterization of the open-field and light/dark tests.

INTRODUCTION In order to assess anxiety in mammals various tests and species are currently available. These current assays measure changes in anxiety-like behaviors. The open-field and the light/dark are anxiety tests based on the spontaneous behavior of the animals, with C57BL/6J mice being a frequently used strain in behavioral studies. However, the suitability of this strain as a choice in anxiety studies has been questioned. In this study, we performed two pharmacological characterizations of this strain in both the open-field and the light/dark tests. METHODS We examined the changes in the anxiety-like behaviors of C57BL/6J mice exposed to chlordiazepoxide (CDP), an anxiolytic drug, at doses of 5 and 10 mg/kg, picrotoxine (PTX), an anxiogenic drug, at doses of 0.5 and 1 mg/kg, and methylphenidate (MPH), a psychomotor stimulant drug, at doses of 5 and 10 mg/kg, in a first experiment. In a second experiment, we tested CDP at 2.5 mg/kg, PTX at 2 mg/kg and MPH at 2.5 mg/kg. RESULTS Results showed an absence of anxiolytic-like effects of CDP in open-field and light/dark tests. Light/dark test was more sensitive to the anxiogenic effects of PTX than the open-field test. Finally, a clear anxiogenic effect of MPH was observed in the two tests. DISCUSSION Although C57BL/6J mice could not be a sensitive model to study anxiolytic effects in pharmacological or behavioral interventions, it might be a suitable model to test anxiogenic effects. Further studies are necessary to corroborate these results.

Impaired Spatial Learning and Unaltered Neurogenesis in a Transgenic Model of Alzheimers Disease After Oral Aluminum Exposure

Although it is well established that aluminum (Al) is neurotoxic, the potential role of this element in the etiology of Alzheimers disease (AD) is not well established. In this study, we evaluated the effects of oral Al exposure on spatial learning, memory and neurogenesis in Tg2576 mice, an animal model of AD in which Abeta plaques start to be deposited at 9 months of age. Aluminum was given as Al lactate (11 mg/g of food) for 6 months. At 11 months of age a water maze test was carried out to evaluate learning and memory. Subsequently, mice were injected with bromo-deoxyuridine (BrdU) and sacrificed 24 hours or 28 days after the last injection in order to assess proliferation, survival and differentiation of neurons. We observed impaired acquisition in the water maze task in Al-treated Tg2576 mice, as well as worse memory in the Al-exposed groups. In terms of neurogenesis, no effects of aluminum were observed in proliferation, survival and differentiation. The results of this investigation suggest that Tg2576 mice fed for 210 days with rodent chow supplemented with Al lactate at 11 mg/g of food have impaired spatial learning although their neurogenesis remains unmodified.

Evaluation of the protective role of melatonin on the behavioral effects of aluminum in a mouse model of Alzheimer's disease.

In this study, five months old female transgenic (Tg2576) and wild-type mice were exposed for 6 months to aluminum (1mg Al/g diet), melatonin (10mg/kg/day), Al plus melatonin, or vehicle only (control group). General motor activity was evaluated using an open-field, whereas spatial learning and memory were assessed in a water maze. Aluminum levels in hippocampus, cortex and cerebellum were determined. Aluminum-treated Tg2576 mice showed higher Al levels in hippocampus than non-Al treated animals. No Al effects on general motor activity were found, while the open-field test showed an increased number of rearings in Tg2576 mice. A lower habituation pattern was observed in melatonin-treated animals. Differences in learning were noted in the water maze acquisition test, in which Al-treated Tg2576 mice showed more difficulties in learning the task than Al-exposed wild mice. No significant effects of melatonin in the acquisition task were observed. The present results indicate that Tg2576 mice are sensitive to high dietary Al levels. A significant protector role of melatonin on Al-induced behavioral effects was not observed.

Amyloid β peptide levels increase in brain of AβPP Swedish mice after exposure to chlorpyrifos.

Chlorpyrifos (CPF) is an organophosphate pesticide widely used in intensive agriculture. Various studies have demonstrated delayed neurotoxic effects in adult mammals after acute CPF exposure. This pesticide induces oxidative stress and neuronal damage, which suggests a possible relationship between CPF exposure and Alzheimers disease (AD). In the present study, we examined in a mice model of AD, long-term changes in the behavior and brain levels of amyloid β after acute CPF exposure. Fifty mg/kg of CPF were subcutaneously injected to Tg2576 (Tg) mice carrying the Swedish amyloid-β protein precursor (AβPP) mutation for AD. General status, body weight, acetyl cholinesterase (AChE) inhibition, and behavioral changes were assessed. Amyloid β fragment (1-40 and 1-42) levels were also measured in the cortical and hippocampal brain regions. A significant and transient decrease in body weight was observed 72 hr after treatment, while no autonomic effects were noted. Motor activity was decreased in Tg mice seven months after CPF treatment. Acquisition learning in a water maze task was not affected, but retention was ameliorated in CPF-exposed Tg mice. Amyloid β levels increased in the brains of treated Tg mice eight months after CPF exposure. The results of this study show that some behavioral changes persisted or emerged months after acute CPF exposure, while amyloid β levels increased. These findings raise concern about the risk of developing neurodegenerative diseases following moderate exposure to CPF in vulnerable subjects.

Behavioral effects of aluminum in mice: influence of restraint stress.

The influence of restraint stress on potential aluminum (Al)-induced behavioral changes was assessed in CD-1 mice. Three groups of adult mice were given 0, 300 and 600 mg Al/kg body weight per day in drinking water for 2 weeks. One-half of the animals in each group were concurrently subjected to restraint stress during 1 h per day throughout the study. After cessation of treatment, open-field activity, active avoidance learning, and motor resistance and coordination of the animals were evaluated. At the end of the behavioral testing period, mice were killed and Al concentrations were determined in a number of tissues. There were no remarkable effects of Al, restraint stress or their combined administration on either open-field activity or on the number of avoidances in an automatic reflex conditioner. However, a lower motor resistance and coordination in a rotarod were observed following exposure to Al at 600 mg/kg/day, restraint alone or concurrent administration of Al (300 and 600 mg/kg/day) plus restraint stress. The levels of Al in whole brain and cerebellum were significantly enhanced in mice exposed to Al plus restraint. Although the present results scarcely show Al-induced neurobehavioral effects, the influence of restraint stress on Al levels in whole brain and cerebellum can be the basis for further studies on the potential role of this element in certain neurological disorders.