Behavioral and biochemical effects of environmental concentrations of caffeine in zebrafish after long-term exposure.

Caffeine (CAF) is widely detected in aquatic environments, serving as an indicator of anthropogenic contamination. Its high consumption, and persistence raise environmental concerns. This study was to evaluate the chronic effects in terms of growth rate, weight, behavior, and biochemical parameters of environmental concentrations of CAF on adult zebrafish. Adult zebrafish were exposed, for 30 d, to 0, 0.5, 1.5, and 300 µg L-1 CAF, with behavior (feeding latency, exploration, aggression, sociability, sound response) and biochemical endpoints (acetylcholinesterase (AChE), lactate dehydrogenase (LDH), and cortisol levels) assessed at the end of the exposure. CAF 0.5 µg L-1 increased feeding latency time, while 300 µg L-1 reduced growth and weight. Exposure to CAF affect fish behavior in terms of vertical exploration, aggressiveness, shoaling, and sound responses although were concentration specific. All concentrations tested increased social behavior, with fish swimming closer to the shoal. At a biochemical level, CAF exposed showed reduced AChE activity, while LDH activity, and cortisol levels increased at 300 µg L-1. Low concentrations of CAF caused neurotoxicity in zebrafish which may compromise their feeding behavior, and social interactions in the wild. These changes suggest potential ecological impacts of chronic exposure to CAF, such as impaired feeding and stress responses.

Effects of environmental concentrations of caffeine on adult zebrafish behaviour: a short-term exposure scenario.

Caffeine (CAF) has been considered an emerging environmental contaminant and its presence indicator of anthropogenic contamination. This study evaluated the effects of environmental concentrations of CAF (0, 0.5, 1.5, and 300 µg. L-1) on the behaviour of adult zebrafish (Danio rerio) after 7 days of exposure. The components of feeding, locomotion, boldness (new tank test), sociability (schooling test), and aggression (mirror test) were analysed. Growth rate and weight were investigated as complementary measures. CAF (0.5, 1.5, and 300 µg. L-1) reduced exploratory behaviour in zebrafish, increased feeding latency time (1.5, and 300 µg. L-1), and decreased growth rate and fish weight (300 µg. L-1). CAF also induced aggressive behaviour (0.5, 1.5, and 300 µg. L-1) and decreased appetence to the shoal (sociability) (0.5, and 1.5 µg. L-1). This study showed that low doses of CAF can induce behavioural effects in zebrafish that may have significant long-term impacts on vital ecological functions.

Early-life metabolic dysfunction impairs cognition and mitochondrial function in mice.

The impact of overnutrition early in life is not restricted to the onset of cardiovascular and metabolic diseases, but also affects critical brain functions related to cognition. This study aimed to evaluate the relationship between peripheral metabolic and bioenergetic changes induced by a two-hit protocol and their impact on cognitive function in juvenile mice. Three-week-old male C57BL/6 mice received a high-fat diet (HFD) or control diet for 7 weeks, associated with two low doses of streptozotocin (STZ) or vehicle. Despite the absence of obesity, HFD+STZ impaired glucose metabolism and induced a trend towards cholesterol increase. The two-hit protocol impaired recognition and spatial memories in juvenile mice, without inducing a depressive-like behavior. HFD+STZ mice presented increased immunoreactivity for GFAP and a trend towards a decrease in NeuN in the hippocampus. The treatment caused a bioenergetic impairment in the hippocampus, characterized by a decrease in both O2 consumption related to ATP production and in the maximum respiratory capacity. The thermogenic capacity of brown adipose tissue was impaired by the two-hit protocol, here verified through the absence of a decrease in O2 consumption after uncoupled protein-1 inhibition and an increase in the reserve respiratory capacity. Impaired mitochondrial function was also observed in the liver of HFD+STZ juvenile mice, but not in their heart. These results indicate that exposure to HFD+STZ early in life has a detrimental impact on the bioenergetic and mitochondrial function of tissues with metabolic and thermogenic activities, which is likely related to hippocampal metabolic changes and cognitive impairment.

Short-term high-fat diet induces cognitive decline, aggression, and anxiety-like behavior in adult zebrafish.

Obesity is a global health problem with high prevalence and defined by a high body mass index (BMI). Several comorbidities affecting the central nervous system (CNS) are associated with obesity (e.g., neurodegenerative diseases, cognitive deficit, and psychobehavioral disturbs). The zebrafish (Danio rerio) has been considered a suitable model organism to investigate the neurobehavioral features of various human diseases. Here, we verify the impact of a high-fat diet (HFD) on the CNS by specifically assessing the effects of short-term HFD on anxiety-like responses, aggression, social preference, and memory, which are essential behaviors for survival and reproduction. Animals were separated in three experimental groups. The standard diet group (SD) received 7.5 mg/fish of dry food, while HFD groups received 5 mg/fish dry food plus 7.5 (HFD-7.5) or 15 mg/fish (HFD-15) of chicken egg yolk daily. Dietary fat content (w/w) was approximately 6.5%, 16.9%, and 21.1%, respectively. We performed behavioral tests and morphometric analyses after two weeks of HFD. In comparison to SD animals, HFD groups showed typical obesogenic responses with increases in BMI, abdominal length, and body weight. HFD individuals also showed increased aggression and anxiety-like behaviors in the mirror-induced aggression and novel tank diving tests, respectively. Interestingly, HFD did not change the social preference behavior, mean swimming speed or spontaneous activity levels, while the HFD-15 group showed cognitive deficits in the inhibitory avoidance test. Collectively, this "proof-of-concept" study is the first report to characterize the effects of short-term HFD on different behavioral domains of zebrafish with high degree of face validity. Moreover, our data reinforce the growing utility of zebrafish to explore the neurobehavioral basis of obesity, providing clinically translatable data, complementing the existing rodent models and supporting future mechanistic studies.

Metabolic activity in cryopreserved and grafted ovarian tissue using high-resolution respirometry.

Cryopreservation of ovarian tissue followed by transplantation represents a strategy to restore ovarian function and fertility. Stress from cryopreservation-thawing processes can lead to alterations and/or damage to mitochondrial structure and functionality. High resolution respirometry and histological analysis were used to evaluate the effect of cryopreservation and transplantation on ovarian tissue. Four different conditions were performed: Fresh non-transplanted tissue, Fresh transplanted tissue, Cryopreserved non-transplanted tissue and Cryopreserved transplanted tissue. All groups were able to respond to the substrates-uncoupler-inhibitor protocol. We found a dramatic decrease in general oxygen consumption in hemi-ovaries submitted to cryopreservation and/or transplantation. The effect of cryopreservation on mitochondrial metabolism was less intense than effect of transplantation, since the transplantation affected all of the mitochondrial states. A total of 2644 follicles were analyzed. Of these, 2198 were classified as morphologically normal. The percentage of morphologically normal follicles was significantly lower in the Cryopreserved transplanted group when compared to the Cryopreserved non-transplanted group and the Fresh transplanted group (p-value < 0.05). Despite decreased follicular viability and mitochondrial activity, the cryopreservation followed by transplantation of ovarian tissue proved feasible for attempts to restore ovarian function.

Potential neuroprotective and anti-inflammatory effects provided by omega-3 (DHA) against Zika virus infection in human SH-SY5Y cells.

Zika virus (ZIKV) has a strong tropism for the nervous system and has been related to post-infection neurological syndromes. Once neuronal cells are infected, the virus is capable of modulating cell metabolism, leading to neurotoxicity and cellular death. The negative effect of ZIKV in neuron cells has been characterized. However, the description of molecules capable of reversing these cytotoxic effects is still under investigation. In this context, it has been largely demonstrated that docosahexaenoic acid (DHA), an omega-3 polyunsaturated fatty acid, is highly neuroprotective. Here, we hypothesized that DHA's neuroprotective proprieties could have an influence on ZIKV-induced neurotoxicity in SH-SY5Y cells. Our data showed that pre-treatment of SH-SY5Y cells with DHA increased the cell viability and proliferation in ZIKV-infected cells. Moreover, DHA triggered an anti-inflammatory response in those infected cells. Besides, DHA was capable of restoring mitochondria function and number in ZIKV-infected SH-SY5Y cells. In addition, cells pre-treated with DHA prior to ZIKV infection presented a lower viral load at different times of infection. Taking together, these results demonstrated that DHA has a potential anti-inflammatory and neuroprotective effect against ZIKV infection in these neuron-like cells and could be a useful tool in the treatment against this virus.