Contrasting neurochemical and behavioral profiles reflects stress coping styles but not stress responsiveness in farmed gilthead seabream (Sparus aurata).

In fish, as well as in other vertebrates, contrasting suites of physiological and behavioral traits, or coping styles, are often shown in response to stressors. However, the magnitude of the response (i.e. stress responsiveness) has been suggested to be independent of stress coping style. One central neurotransmitter that has been associated with both stress responsiveness and differences in stress coping styles is serotonin (5-hydroxytryptamine, 5-HT). In this study, we investigated to what extent stress responsiveness reflects differences in stress coping, and the potential involvement of the 5-HT system in mediating such differences in farmed Gilthead seabream. Initially, fish were classified as proactive or reactive based on their behavioural response to net restraint. Following 1.5 months, fish classified as proactive still showed a higher number of escape attempts and spent longer time escaping than those classified as reactive. These differences were reflected in a generally higher brain stem 5-HT concentration and a lower telencephalic 5-HT activity, i.e. the ratio of 5-hydroxyindoleacetic acid (5-HIAA) to 5-HT, in proactive fish. Independent of stress coping styles, stress responsiveness was reflected in elevated 5-HIAA concentrations and 5-HIAA/5-HT ratios in telencephalon and brain stem together with increased plasma cortisol concentrations at 0.5 and 2 h following the last net restraint. The current results show that 5-HT signaling can reflect different behavioural output to a challenge which are independent of neuroendocrine responses to stress and lend support to the hypothesis that stress coping styles can be independent of stress responsiveness.

Contrasting Coping Styles Meet the Wall: A Dopamine Driven Dichotomy in Behavior and Cognition.

Individual variation in the ability to modify previously learned behavior is an important dimension of trait correlations referred to as coping styles, behavioral syndromes or personality. These trait clusters have been shaped by natural selection, and underlying control mechanisms are often conserved throughout vertebrate evolution. In teleost fishes, behavioral flexibility and coping style have been studied in the high (HR) and low-responsive (LR) rainbow trout lines. Generally, proactive LR trout show a behavior guided by previously learned routines, while HR trout show a more flexible behavior relying on environmental cues. In mammals, routine dependent vs. flexible behavior has been connected to variation in limbic dopamine (DA) signaling. Here, we studied the link between limbic DA signaling and individual variation in flexibility in teleost fishes by a reversal learning approach. HR/LR trout were challenged by blocking a learned escape route, previously available during interaction with a large and aggressive conspecific. LR trout performed a higher number of failed escape attempts against the transparent blockage, while HR trout were more able to inhibit the now futile escape impulse. Regionally discrete changes in DA neurochemistry were observed in micro dissected limbic areas of the telencephalon. Most notably, DA utilization in the dorsomedial telencephalon (DM, a suggested amygdala equivalent) remained stable in HR trout in response to reversal learning under acute stress, while increasing from an initially lower level in LR trout. In summary, these results support the view that limbic homologs control individual differences in behavioral flexibility even in non-mammalian vertebrates.

A novel role for pigment genes in the stress response in rainbow trout (Oncorhynchus mykiss).

In many vertebrate species visible melanin-based pigmentation patterns correlate with high stress- and disease-resistance, but proximate mechanisms for this trait association remain enigmatic. Here we show that a missense mutation in a classical pigmentation gene, melanocyte stimulating hormone receptor (MC1R), is strongly associated with distinct differences in steroidogenic melanocortin 2 receptor (MC2R) mRNA expression between high- (HR) and low-responsive (LR) rainbow trout (Oncorhynchus mykiss). We also show experimentally that cortisol implants increase the expression of agouti signaling protein (ASIP) mRNA in skin, likely explaining the association between HR-traits and reduced skin melanin patterning. Molecular dynamics simulations predict that melanocortin 2 receptor accessory protein (MRAP), needed for MC2R function, binds differently to the two MC1R variants. Considering that mRNA for MC2R and the MC1R variants are present in head kidney cells, we hypothesized that MC2R activity is modulated in part by different binding affinities of the MC1R variants for MRAP. Experiments in mammalian cells confirmed that trout MRAP interacts with the two trout MC1R variants and MC2R, but failed to detect regulation of MC2R signaling, possibly due to high constitutive MC1R activity.

Stress and fear responses in the teleost pallium.

Evolution has resulted in behavioural responses to threat which show extensive similarities between different animal species. The reaction to predator cues is one example of such prevailing responses, and functional homologies to mammalian limbic regions involved in threat-sensitive behaviour have been found in the teleost telencephalon. The dorsolateral (Dl) and dorsomedial (Dm) regions of the pallium are thought to perform hippocampus and amygdala-like functions respectively. To what degree these regions are involved in the neuroendocrine responses to stress and predator cues however remains largely unknown. In the present study the involvement of Dl and Dm in such responses was investigated by exposing Nile tilapia (Oreochromis niloticus) to a standardized confinement stress and to skin extract from conspecifics. Nile tilapia develops a characteristic anticipatory behaviour to hand feeding, and effects of skin extract on this behaviour and locomotor activity were studied to characterise threat sensitive behaviour. Nile tilapia responded behaviourally to conspecific alarm cues by reducing feeding anticipatory behaviour. This may reflect a general elevation of alertness, and further studies combining skin extract with other challenges are needed to reveal neuroendocrine effects associated with this predator cue. Confinement stress resulted in an elevation of cortisol and serotonin (5-hydroxytryptamine, 5-HT) metabolism in both Dl and Dm. A similar tendency was observed in fish exposed to chemical alarm cues, but this effect did not reach the level of statistical significance. Hence, limbic responses to stress and fear, akin to those seen in extant mammals, are also present in the teleost lineage.

Feeding motivation as a personality trait in Nile tilapia (Oreochromis niloticus): role of serotonergic neurotransmission.

Consistent individual variation in behaviour and physiology (i.e. animal personality or coping style) has emerged as a central topic in many biological disciplines. Yet, underlying mechanisms of crucial personality traits like feeding behaviour in novel environments remain unclear. Comparative studies, however, reveal a strong degree of evolutionary conservation of neural mechanisms controlling such behaviours throughout the vertebrate lineage. Previous studies have indicated duration of stress-induced anorexia as a consistent individual characteristic in teleost fishes. This study aims to determine to what degree brain 5-hydroxytryptamine (5-HT, serotonin) activity pertains to this aspect of animal personality, as a correlate to feed anticipatory behaviour and recovery of feed intake after transfer to a novel environment. Crucial to the definition of animal personality, a strong degree of individual consistency in different measures of feeding behaviour (feeding latency and feeding score), was demonstrated. Furthermore, low serotonergic activity in the hypothalamus was highly correlated with a personality characterized by high feeding motivation, with feeding motivation represented as an overall measure incorporating several behavioural parameters in a Principle Component Analyses (PCA). This study thus confirms individual variation in brain 5-HT neurotransmission as a correlate to complex behavioural syndromes related to feeding motivation.

High oxygen consumption rates and scale loss indicate elevated aggressive behaviour at low rearing density, while elevated brain serotonergic activity suggests chronic stress at high rearing densities in farmed rainbow trout.

The effect of stocking density on indicators of welfare has been investigated by several studies on farmed rainbow trout Oncorhynchus mykiss. However, the densities at which welfare are compromised remain ambiguous. Here three different stocking density treatments were selected based on the results of a previous study, where levels of crowding where determined using the spatial distribution of fish in two-tank systems. An un-crowded low density of 25 kgm(-3), the highest density accepted by the fish without showing indications of crowding stress of 80 kgm(-3) as the intermediate density, and the highest density accepted by the fish showing indications of crowding stress of 140 kgm(-3) as the high density were investigated. The aim of the present study was to examine the effect of being held at these densities on indicators of welfare. This was achieved through oxygen consumption measurements using automated respirometry, recording fin erosion, determining scale loss and analysing plasma cortisol and brain serotonergic activity levels. The results obtained in the present study indicated that at the lowest density the fish had the space and opportunity to display their natural aggressive behaviour and that the fish held at the highest density were exposed to a situation of confinement.

Linking fearfulness and coping styles in fish.

Consistent individual differences in cognitive appraisal and emotional reactivity, including fearfulness, are important personality traits in humans, non-human mammals, and birds. Comparative studies on teleost fishes support the existence of coping styles and behavioral syndromes also in poikilothermic animals. The functionalist approach to emotions hold that emotions have evolved to ensure appropriate behavioral responses to dangerous or rewarding stimuli. Little information is however available on how evolutionary widespread these putative links between personality and the expression of emotional or affective states such as fear are. Here we disclose that individual variation in coping style predicts fear responses in Nile tilapia Oreochromis niloticus, using the principle of avoidance learning. Fish previously screened for coping style were given the possibility to escape a signalled aversive stimulus. Fearful individuals showed a range of typically reactive traits such as slow recovery of feed intake in a novel environment, neophobia, and high post-stress cortisol levels. Hence, emotional reactivity and appraisal would appear to be an essential component of animal personality in species distributed throughout the vertebrate subphylum.