The growing and continuous use of drugs by the human population transformed the pharmacological agents in "emerging pollutants" are often found in aquatic environments. The consequences of this finding are still unclear, and little is known about the effects of these compounds on the environment and the health of exposed species. The excretion of these components still in its active form and the improper disposal thereof are among the main causes of environmental contamination. The use of embryos and larvae of zebrafish (Danio rerio) as an experimental model for toxicological testing has shown a number of benefits when compared to other animal models, as they represent a low cost model, easily handled, fast reproduction. In addition the extracorporeal development and embryonic transparency of zebrafish make it an ideal alternative to determine of potential lethality and teratogenicity of different molecules. The aim of the study was to evaluate the effects caused by exposure to the psychiatric drugs diazepam, fluoxetine and risperidone on embryos and larvae of zebrafish, taking into account survival parameters, hatching, heart rate and overall length. The embryos were exposed to different concentrations of each drug, taking into account concentrations already found in the aquatic environment and described in the literature. Risperidone, fluoxetine and diazepam affect the early development of zebrafish, the first drug having greater effect, causing changes in all evaluated parameters. When placed in an environmental context, the effects can interfere with population levels and survival of the species in natural environment. / O uso crescente e continuo de fármacos pela população humana fazem dos agentes farmacológicos potentes poluentes emergentes e são frequentemente encontrados em ambientes aquáticos. A consequência destes achados ainda não é esclarecida, e pouco se sabe sobre os efeitos desses compostos sobre o ambiente e a saúde das espécies expostas. A excreção desses componentes ainda em sua forma ativa e o descarte inadequado dos mesmos estão entre as principais causas de contaminação ambiental. O uso de embriões e larvas de zebrafish (Daniorerio) como modelo experimental para ensaios toxicológicos vem mostrando uma série de benefícios quando comparados á utilização de outros modelos animais, já que representam um modelo de baixo custo, fácil manejo e reprodução. Além disso o desenvolvimento extracorpóreo e a transparência embrionária fazem do zebrafish uma alternativa ideal para determinação de potenciais de letalidade, subletalidade e teratogenicidade. O objetivo do estudo foi avaliar os efeitos provocados pela exposição aos psicofármacosdiazepam, fluoxetina e risperidonasobre embriões e larvas de zebrafish, levando em consideração os parâmetros sobrevivência, eclosão, frequência cardíaca e comprimento total. Os embriões foram expostos a diferentes concentrações de cada fármaco, utilizando como base as concentrações já encontradas no ambiente aquático e descritas na literatura. A risperidona, a fluoxetina e o diazepam afetam o desenvolvimento inicial do zebrafish, o primeiro fármaco promovendo maior efeito, provocando alterações nos quatro parâmetros avaliados. Quando colocados em um contexto ambiental, os efeitos encontrados podem interferir os níveis populacionais e manutenção da espécie em ambiente natural.

Fluoxetine (FLX), the active ingredient of Prozac™, is a member of the selective serotonin reuptake inhibitor (SSRI) class of anti-depressant drugs and is present in aquatic environments worldwide. Previous studies reported that FLX is an endocrine disruptor in fish, bioconcentrating in tissues including the brain. Evidence implicates that serotonin influences the activity of the hypothalamo-pituitary-interrenal (HPI) stress axis, thus exposure to FLX may disrupt the teleost stress response. This study examined in vitro cortisol production in rainbow trout (Oncorhynchus mykiss) head kidney/interrenal cells exposed to FLX and 14C-pregnenolone metabolism in head kidney microsome preparations of FLX-exposed trout. Results indicated that cells exposed in vitro to increasing concentrations of FLX had lower cortisol production and cell viability (versus control) and microsomes isolated from trout exposed to 54 μg/L FLX had higher pregnenolone metabolism versus those of control and low FLX-exposed (0.54 μg/L) trout.

Fluoxetine, a selective serotonin re-uptake inhibitor (SSRI) and the active ingredient in Prozac®, is found in the environment and disrupts feeding and metabolism in exposed fish. The objective of this research was to investigate the mechanisms involved in the feeding and metabolism disruption in the model goldfish (Carassius auratus). Two short-term waterborne fluoxetine exposures (7- and 14-days) were performed using two environmentally relevant doses of fluoxetine (0.5 and 1 μg/L) and metabolic effects at the level of the brain, liver, serum and bile in goldfish were investigated. Abundances of mRNA transcripts coding for six feeding neuropeptides were examined to determine which may be involved in the initial neural changes associated with decreased appetite in goldfish. The 7-day fluoxetine exposure at 1 μg/L caused corticotropin-releasing factor (CRF) mRNA levels to increase by 2-fold in female hypothalamus and telencephalon, indicating that CRF may be one of the first of the feeding neuropeptides to be altered. Six hepatic miRNAs were also evaluated in the goldfish liver that were previously associated with fluoxetine exposure in zebrafish (Danio rerio). Following the 7-day exposure at 1 μg/L, miR-22b, miR-140, miR-210, miR-301a and miR-457b levels increased in the female goldfish liver by 4-6 fold. The 14-day fluoxetine exposure at 1 μg/L caused 2-fold increases in miR-210, miR-301a, miR-457b and let-7d in male goldfish liver. These miRNAs were associated with the down-regulation of anabolic metabolic pathways in zebrafish, indicating a conservation of miRNA and fluoxetine effect between fish species. Serum and bile metabolite profiles of fluoxetine exposed goldfish were evaluated using ultra performance liquid chromatography coupled to quadrupole time of flight mass spectrometry. Following the 14-day exposure at 1 μg/L, the bile metabolite profiles of male goldfish were significantly different from controls as detected by cluster analysis and fluoxetine was tentatively identified in the serum. No other discriminant metabolites were identified as of yet. The data presented suggest that fluoxetine causes metabolic disruption in goldfish at multiple organ levels. Because of the widespread detection of fluoxetine and other emerging SSRIs in the aquatic environment, future research is required to firmly establish this pharmaceutical class as a metabolic and endocrine disrupting chemical.

No description available.

Selective serotonin reuptake inhibitors (SSRIs), like fluoxetine, are widely used to treat depressive disorders during pregnancy. These antidepressants reach water reservoirs through sewage treatment facilities and expose the aquatic vertebrates, including fish. It has been shown that early-life exposure to fluoxetine could disrupt the normal function of the stress axis by decreasing the level of circulating glucocorticoids in humans, rodents, and teleosts. Our lab recently showed that early life exposure to fluoxetine resulted in transgenerational hyporcortisolism and altered exploratory behaviour in adult male zebrafish and their descendant male adults for at least three generations. In the current study, we used a stress-responsive transgenic zebrafish line (SR4G) that expresses green fluorescence protein (eGFP) under the control of six consecutive glucocorticoid response elements. The effects of developmental exposure to fluoxetine on the transcriptional profiles of genes in the larval head and male adult telencephalon and hypothalamus were analysed using high throughput RNA sequencing. We also assessed the potential of eGFP mRNA to evaluate blunted stress response as an alternative to cortisol immunoassay measurements. The effects of bisphenol A, vinclozolin and fluoxetine were ytested in the SR4G line. Developmental exposure to fluoxetine resulted in a life-long dysregulation of pathways involved in nervous system development, stress response, and lipid metabolism in both larvae and adult zebrafish. Numerous differentially expressed genes in zebrafish are orthologous to genes in Homo sapiens linked the development of the major depressive disorder and epigenetics regulation and include bdnf, trkb, npas4, per1, per2, dnmt3a, adarb1, adaeb2, hdac4, hdac5, hdac8, and atf2. It is suggested that the dysregulation of the primary transcription regulators of circadian rhythm (clocka) and stress response (nr3c1), amongst others, were the potential drivers of the observed life-long effects. Furthermore, we report on a significant positive linear correlation between cortisol levels and eGFP mRNA levels in SR4G transgenic zebrafish larvae (R2> 0.9). Random forest and logistic regression models trained by eGFP mRNA levels both correctly predicted the blunted stress response. The negative predictive value (NPV) for both models was 100%. Models based on the mRNA levels of 11 genes associated with neurogenesis, stress response and depression resulted in a similar 100% NPV. These findings provide evidence for a life-long effect of developmental exposure to fluoxetine. This study also provides a proof-of-concept for an in vivo biomonitoring assay to screen chemicals for their stress-disrupting potential.

Rationale Many studies demonstrate down-regulation of the norepinephrine transporter (NET) by desipramine (DMI) in vitro and in stress-naive rats. Little is known regarding regulation of the NET in stressed animals. Objective The present study was designed to investigate effects of DMI on the expression of NET and protein kinases in the stress rat. Methods Adult Fischer 344 rats were subjected to chronic social defeat (CSD) for 4 weeks. DMI (10 mg/kg, intraperitoneal (i.p.)) was administered concurrently with CSD or 1 or 2 weeks after cessation of CSD. Sucrose consumption, NET expression, and protein kinases were measured. Results CSD significantly increased messenger RNA (mRNA) and protein levels of NET in the locus coeruleus, as well as NET protein levels in the hippocampus, frontal cortex, and amygdala. These effects were nearly abolished when DMI was administered concurrently with CSD. CSD-induced upregulation of NET expression in the locus coeruleus, hippocampus, and amygdala lasted at least 2 weeks after cessation of CSD, an effect that was significantly attenuated by 1 or 2 weeks of DMI treatment starting from cessation of the CSD. Concurrent administration of DMI with CSD did not markedly interfere with CSD-induced decreases in protein levels of protein kinases A and C in these brain regions, but it did reverse the CSD-induced reduction in phosphorylated cAMP response element-binding (pCREB) protein levels in most brain regions. Conclusion These findings suggest that NET regulation by DMI occurs in both stressed and behaviorally naive rats, and DMI-induced changes in pCREB may be involved.

This study examines the effects of the selective serotonin reuptake inhibitor (SSRI), fluoxetine (PROZAC), on the ontogeny of spontaneous swimming activity (SSA) in developing zebrafish. The development of zebrafish motor behavior consists of four sequential locomotor patterns that develop over 1-5 days post fertilization (dpf), with the final pattern, SSA, established at 4-5 dpf. In stage specific experiments, larvae were exposed to 4.6 μM fluoxetine for 24 h periods beginning at 24 h post fertilization (hpf) and extending through 5 dpf. From 1-3 dpf, there was no effect on SSA or earlier stages of motor development, i.e., spontaneous coiling, evoked coiling and burst swimming. Fluoxetine exposure at 3 dpf for 24 h resulted in a transient decrease in SSA through 7 dpf with a complete recovery by 8 dpf. Larvae exposed to 4.6 μM fluoxetine for 24 h on 4 or 5 dpf showed a significant decrease in SSA by day 6 with no recovery through 14 dpf. Although SSA was significantly affected 24 h after fluoxetine exposure, there was little or no effect on pectoral fin movement. These results demonstrate both a stage specific and a long term effect of 4.6 μM fluoxetine exposure in 4 and 5 dpf larvae. Reverse transcriptase polymerase chain reaction (RT-PCR) was performed to determine the relative levels of a serotonin transporter protein (SERT) transcript and the serotonin 1A (5-HT1A) receptor transcript in developing embryos/larvae over 1-6 dpf. Both transcripts were present at 24 hpf with the relative concentration of SERT transcript showing no change over the developmental time range. The relative concentration of the 5-HT1A receptor transcript, however, showed a two-tiered pattern of concentration. RT-PCR was also used to detect potential changes in the SERT and 5-HT1A receptor transcripts in 6 dpf larvae after a 24 h exposure to 4.6 μM fluoxetine on 5 dpf. Three separate regions of the CNS were individually analyzed, two defined brain regions and spinal cord. The two brain regions showed no effect on transcript levels subsequent to fluoxetine exposure, however, the spinal cord showed a significant decrease in both transcripts. These results suggest a correlation between decreased concentration of SERT and 5-HT1A receptor transcripts in spinal cord and decreased SSA subsequent to fluoxetine exposure.

No description available.

No description available.

No description available.