R. van den Bos

Unpredictable chronic stress decreases inhibitory avoidance learning in Tuebingen long-fin zebrafish: stronger effects in the resting phase than in the active phase

Zebrafish (Danio rerio Hamilton) are increasingly used as a model to study the effects of chronic stress on brain and behaviour. In rodents, unpredictable chronic stress (UCS) has a stronger effect on physiology and behaviour during the active phase than during the resting phase. Here, we applied UCS during the daytime (active phase) for 7 and 14 days or during the night-time (resting phase) for 7 nights in an in-house-reared Tuebingen long-fin (TLF) zebrafish strain. Following UCS, inhibitory avoidance learning was assessed using a 3 day protocol where fish learn to avoid swimming from a white to a black compartment where they will receive a 3 V shock. Latencies of entering the black compartment were recorded before training (day 1; first shock) and after training on day 2 (second shock) and day 3 (no shock, tissue sampling). Fish whole-body cortisol content and expression levels of genes related to stress, fear and anxiety in the telencephalon were quantified. Following 14 days of UCS during the day, inhibitory avoidance learning decreased (lower latencies on days 2 and 3); minor effects were found following 7 days of UCS. Following 7 nights of UCS, inhibitory avoidance learning decreased (lower latency on day 3). Whole-body cortisol levels showed a steady increase compared with controls (100%) from 7 days of UCS (139%), to 14 days of UCS (174%) to 7 nights of UCS (231%), suggestive of an increasing stress load. Only in the 7 nights of UCS group did expression levels of corticoid receptor genes (mr, grα, grβ) and of bdnf increase. These changes are discussed as adaptive mechanisms to maintain neuronal integrity and prevent overload, and as being indicative of a state of high stress load. Overall, our data suggest that stressors during the resting phase have a stronger impact than during the active phase. Our data warrant further studies on the effect of UCS on stress axis-related genes, especially grβ; in mammals this receptor has been implicated in glucocorticoid resistance and depression.

Switching to cue-directed behavior: specific for ventral striatal dopamine but not ventral pallidum/substantia innominata gaba as revealed by a swimming-test procedure in rats

In this study it was investigated whether ventral striatal dopamine-induced changes in switching to cue-directed behavioral patterns were funnelled via the rostral areas of the ventral pallidum/substantia innominata (VP/SI) complex and, if so, whether changes in switching to cue-directed behavioral patterns could be elicited in the VP/SI complex by manipulating GABAergic activity. To this end rats were bilaterally equipped with cannulae directed at the ventral striatum and/or rostral VP/SI complex and subjected to a swimming-test procedure for 6 min. Injections of the dopamine-releasing agent d-amphetamine (10 microg/0.5 microl per side) enhanced the number of different cue-directed behavioral patterns while they had no effect upon the number of different non-cue-directed behavioral patterns in line with previous studies (Life Sci - 1989 1697). This increase was attenuated by a low dose of the GABAa agonist muscimol (1 ng/0.5 microl) into the rostral VP/SI complex. This dose of muscimol when injected alone into the rostral VP/SI complex had no effect upon the number of different cue-directed behavioral patterns. Only the lowest dose of the GABAa antagonist bicuculline (10-25 ng/0.5 microl per side) into the rostral VP/SI complex slightly, and in a non-d-amphetamine-like manner, increased the number of different cue-directed behavioral patterns while none of the doses had an effect on the number of different non-cue-directed behavioral patterns. Both injections of d-amphetamine into the ventral striatum and injections of bicuculline into the rostral VP/SI complex strongly increased motor activity in the 10-min period preceding the swimming test. We conclude from the data that switching to cue-directed behavioral patterns is sensitive to manipulations with the dopaminergic activity in the ventral striatum but not with the GABAergic activity in the VP/SI complex although the VP/SI transmits it to other brain structures. In contrast motor activity is sensitive to manipulations with both ventral striatal dopamine and rostral VP/SI complex GABA.

The effects of environmental enrichment and age-related differences on inhibitory avoidance in zebrafish (Danio rerio Hamilton).

The inhibitory avoidance paradigm allows the study of mechanisms underlying learning and memory formation in zebrafish (Danio rerio Hamilton). For zebrafish, the physiology and behavior associated with this paradigm are as yet poorly understood. We therefore assessed the effects of environmental enrichment and fish age on inhibitory avoidance learning. Fish raised in an environmentally enriched tank showed decreased anxiety-like behavior and increased exploration. Enrichment greatly reduced inhibitory avoidance in 6-month (6M)- and 12-month (12 M)-old fish. Following inhibitory avoidance, telencephalic mRNA levels of proliferating cell nuclear antigen (pcna), neurogenic differentiation (neurod), cocaine- and amphetamine-regulated transcript 4 (cart4), and cannabinoid receptor 1 (cnr1) were lower in enriched-housed fish, while the ratios of mineralocorticoid receptor (nr3c2)/glucocorticoid receptor α [nr3c1(α)] and glucocorticoid receptor β [nr3c1(β)]/glucocorticoid receptor α [nr3c1(α)] were higher. This was observed for 6M-old fish only, not for 24-month (24 M) old fish. Instead, 24 M-old fish showed delayed inhibitory avoidance, no effects of enrichment, and reduced expression of neuroplasticity genes. Overall, our data show strong differences in inhibitory avoidance behavior between zebrafish of different ages and a clear reduction in avoidance behavior following housing under environmental enrichment.

Differences in inhibitory avoidance, cortisol and brain gene expression in TL and AB zebrafish.

Recently, we established an inhibitory avoidance paradigm in Tupfel Long‐Fin (TL) zebrafish. Here, we compared task performance of TL fish and fish from the AB strain; another widely used strain and shown to differ genetically and behaviourally from TL fish. Whole‐body cortisol and telencephalic gene expression related to stress, anxiety and fear were measured before and 2 h post‐task. Inhibitory avoidance was assessed in a 3‐day paradigm: fish learn to avoid swimming from a white to a black compartment where a 3V‐shock is given: day 1 (first shock), day 2 (second shock) and day 3 (no shock, sampling). Tupfel Long‐Fin fish rapidly learned to avoid the black compartment and showed an increase in avoidance‐related spatial behaviour in the white compartment across days. In contrast, AB fish showed no inhibitory avoidance learning. AB fish had higher basal cortisol levels and expression levels of stress‐axis related genes than TL fish. Tupfel Long‐Fin fish showed post‐task learning‐related changes in cortisol and gene expression levels, but these responses were not seen in AB fish. We conclude that AB fish show higher cortisol levels and no inhibitory avoidance than TL fish. The differential learning responses of these Danio strains may unmask genetically defined risks for stress‐related disorders.

Effects of acute stress on aggression and the cortisol response in the African sharptooth catfish Clarias gariepinus: differences between day and night.

African sharptooth catfish Clarias gariepinus were housed under continuous dim light (1 lx) or 12L:12D (350-0 lx) cycles. The number of skin lesions, as indicator of aggressive acts, and plasma cortisol levels, as indicator of stress-axis activity, were measured at baseline as well as following a stressor (given in the light or dark phase). Results showed that (1) baseline plasma cortisol levels were not different between photoperiods, (2) the number of baseline skin lesions was highest for C. gariepinus housed under continuous dim light, (3) stressor-induced peak levels of plasma cortisol were highest in the light phase and (4) the number of skin lesions following a stressor was highest in the dark phase. The higher number of stressor-related skin lesions in the dark (active) phase suggests increased stressor-induced aggression while in the active phase. In addition, the data suggest that housing under continuous dim light does not result in higher stress-axis activity, as measured by baseline levels of cortisol, but does result in more stressor-induced aggression, as measured by the higher number of skin lesions. The latter may be related to the fact that the continuous dim light photoperiod has twice the number of dark-phase (active) hours in which stressor-induced aggression is stronger compared to the 12L:12D photoperiod, which has a light phase in which stressor-induced aggression is lower.

The ‘ins and Outs’ of Sex Differences in Pathological Gambling

Decision-making plays a pivotal role in daily life as impairments in processes underlying decision-making often result in an inability to make profitable long-term decisions that incorporate expectations of future outcomes. In case of pathological gambling such impairments appear to be present as subjects continue to gamble despite the fact that this activity disrupts their personal, professional or financial life. Both from a research and treatment point of view, an important issue is whether differences between men and women exist in the tendency to engage in gambling as a leisure activity and to develop pathological gambling. I will discuss that differences between male and female subjects may exist regarding the trajectory towards pathological gambling: when considered in aggregate, there appears to be a (slightly) elevated risk for gambling problems or severity of gambling in men compared to women. These differences seem to be due to a complex interaction of genetic make-up, effects of the (social) environment on risk-taking behaviour, stress-related changes in risk-taking, and personality/information-processing characteristics related to decision-making. These different factors will be discussed. Future studies should address the interaction between these factors using among others ecologically valid research methods.