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Debate surrounds the role of behaviour in animal physiology, divided between traditional views that separate behaviour and physiology and a progressive perspective that supports the notion of behaviour as an inherent physiological effector. We advocate for the latter, and in this poster we provide a mechanistic and motivational perspective on behaviour and draw parallels with recognised physiological processes. Indeed, behaviour is ultimately generated by spatial and temporal patterns of musculoskeletal activity and it is tightly regulated by neural pathways, like other physiological processes. For example, many bird species will employ vasoconstriction and shivering – two well-established physiological processes in thermoregulation – and social huddling behaviour to avoid hypothermia. Yet, analogous neuroendocrine patterns can be observed in all three of these physiological actions. Furthermore, these responses are elicited by an animal’s motivational state in response to challenges in its environment. This motivational state can trigger a variety of physiological responses – behavioural or otherwise – that, while operating concurrently, all aim to satisfy an underlying ‘need’. For example, when an animal’s water balance approaches critical levels, it may employ a suite of physiological actions/strategies to conserve (e.g., water reabsorption in the gut, behavioural thermoregulation) and acquire water (e.g., water vapour absorption, drinking/feeding), driven by a motivational state to avoid osmotic stress. Our goal with this poster is to set the stage for a debate and persuade physiologists of the merits of including behaviour within the domain of animal physiology. Recognising behaviour as a physiological process is crucial for advancing a unified understanding of physiology.

SEB Conference 2024

Scaling organismal physiology: a case for the integration of behaviour as a physiological process

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An important trait of Pacific bluefin tuna (PBT) is its ability to maintain body temperature through high metabolic heat production and its retention ability. PBT rapidly develop the thermo-conservation ability between 20 and 40 cm fork length (FL), which has been attributed to improved insulation. Meanwhile, how heat production capacity develops during ontogeny is not well understood. The aim of the present study was to explore the ontogenetic change in metabolic heat and its contribution to endothermic ability through swim-tunnel respirometry and a heat-budget model. Swim-tunnel respirometry was conducted on PBT, ranging from 19.4 to 27.5 cm FL (94–330 g, n=16) in Kochi Prefecture, Japan, from August to September 2022 and 2023. The metabolic rate of each PBT was calculated as the standard metabolic rate (BSMR) at a swim speed of 0 cm/s and the minimum metabolic rate (BUmin) at a minimum swim speed of 44.3 cm/s. The scaling exponents of BSMR and BUmin were estimated at 1.03 and 0.93, respectively. Both scaling exponents of metabolic rates were higher than those estimated for the tribe Thunnini above 300 g. Furthermore, a scaling exponent close to 1 implies that heat production rate (Tm in °C/min) in heat-budget model does not decrease in this body size range. To explore the ontogenetic change of heat-production rate of PBT in nature, heat-budget model was applied to bio-logging data collected from nine juveniles (size of analysis: 23–60 cm FL; 200–5272 g) with electronic archival tags inserted into their body cavity. The heat-production rate of each PBT did not decrease until approximately 800 g and declined thereafter, supporting the results of swim-tunnel respirometry. Furthermore, we evaluated the effects of heat production rate on the thermo-conservation ability, the difference between body temperature and water temperature, and found that maintaining a high heat production rate mainly contributed to the initial stage of temperature difference generation. Our results provided new insights into the endothermal development of tuna.

Highly maintained heat production capacity contributes to the initial step of endothermal development in Pacific bluefin tuna

A great proportion of people with Rett syndrome (RTT) have aberrant pain responses. However, due to additional RTT-related symptoms such as mobility impairments and altered verbal skills, it is difficult to assess their noxious responses. Apart of being considered a pathological state per se, altered pain responses could result in undertreatment of additional RTT-associated conditions. Here, we used male mice null for Mecp2, the gene causing most cases of RTT, to evaluate their response to an inflammatory model. At symptomatic ages (8 weeks old), two cohorts of animals received an intraplantar injection of Complete’s Freud Adjuvant. We analysed either mechanical or thermal sensitivity (von Frey hairs or Hargreaves apparatus, respectively) during basal conditions and at 1- and 3-days post-injection. An hour after last testing, animals were sacrificed and nervous tissues collected for testing. Controls included wild-type (WT) siblings and saline-injected animals from both genotypes. During basal conditions, Mecp2-null mice showed a significantly higher threshold to withdraw their paws in response to a mechanical stimulation than WT and thus are described as mechanically hyposensitive. Interestingly, although hypersensitivity also developed in the mutants at 24 hours post-injection, it was completely resolved by the third day. Immunofluorescence in sectioned brains revealed alterations in the endocannabinoid system within the Mecp2-null mice when compared to WT. Taking all together, our data suggests that MeCP2 is key player in the maintenance, but not the development, of mechanical hypersensitivity following an injury. We also point at the endocannabinoid system as a potential therapeutic target for RTT

Mecp2-null male mice are mechanically hyposensitive before and after an inflammatory pain model

Extreme heat events are predicted to be more frequent and intense over the next few decades. Ectotherms are particularly sensitive to such events due to their reliance on the external environment to perform their daily activities and maintain body temperature. Exposure to higher temperatures during fish development can accelerate growth rates, which directly impacts growth and mortality rates. Yet very little is known about the impact of rearing density and social cues on the behavior of developing embryos exposed to thermal stress. In this study, we measured the startle response of 1 day post fertilization zebrafish embryos at different rearing densities (10, 20, and 40 eggs), and with and without regular social and heat-induced chemical cues. Our results show that activity increases with higher embryo densities, and heat-induced social cues modulate activity both with and without application of thermal stress. Since the startle response is a proxy for how well sensory information and motor actions are coordinated, rearing density and social cues emerge as important factors for understanding the response of developing fish to environmental stressors.

Exploring Zebrafish Embryo Behavioral stress Responses in social contexts

Rett syndrome (RTT) is a neurodevelopmental disorder characterised by, among others, motor impairments, neuroendocrine dysregulation, anxiety altered responses and aberrant pain perception. The primary cause of RTT is de novo loss-of-function mutation in the methyl-CpG-binding domain protein 2 (MECP2) gene, which is located within chromosome X. Although RTT affects mainly females (heterozygous), most rodent studies have used males (hemizygous) as they develop the RTT-associated symptomatology sooner than females (8 weeks vs. 6 months). Here, we analysed both mutant males and females from a CD1 background line at corresponding symptomatic ages, together with control wild-type (WT) siblings. After weighting the animals, they were subjected to an elevated plus maze (EPM) to assess their anxiety responsiveness. One week later, animals received an intraplantar injection of capsaicin to elicit a nociception-specific activation. One hour post-injection, subjects were sacrificed and nervous system collected to assess differential activation in the nociceptive pathway via cFOS immunohistochemistry. Our results show that, although Mecp2 mutant males moved less over the duration of the EPM, they demonstrate a more anxiolytic phenotype than their WT siblings. Mecp2-het females seem to describe a similar behavioural pattern. Nonetheless, mutant females were significantly heavier than their WT siblings while males showed no difference in weight. Following the capsaicin injection, similar changes were observed in both males and females. Together, our results validate this rodent model for the study of RTT-associated neurological manifestation and further demonstrate the need to include animals from both sexes to study the aetiology of RTT and associated disorders.

Mecp2-mutant CD1 mice recapitulate anxiety- and nociceptive-alterations associated with Rett Syndrome

Mice rely on olfactory and vomeronasal marks to recognise other individuals, which need to be integrated into the spatial map to encode the identity of other conspecifics in a context-dependent manner. Our objective is to explore the influence of vomeronasal information in the spatial map characterising the functional interconnection between the chemosensory amygdala and the hippocampus.
We assessed vomeronasal influence on spatial mapping by presenting male urine to female mice in two exploratory behaviours while recording single-unit activity in the hippocampus. In one experiment, mice explore a virtual labyrinth divided into sectors with specific visual or olfactory contingencies. In the other, we designed a test to study place preference and spatial maps simultaneously, the Enriched Open Field (EOF), where we placed modular pieces in an Open Field to enhance exploration and provide references, and including urine marks in a specific quadrant.
The neuronal activity associated with the exploration of the labyrinth showed specific responses to given sectors. These responses include both increased and decreased firing rate usually in the sector where male urine is delivered. In the EOF, in contrast to our expectations, female mice avoid the area where the male left marks. But regarding the neuronal activity, we could identify neurons whose response is associated with male-marked areas.
These experiments broaden our knowledge about how neuronal activity links both exploration patterns and specific locations in space in the context of the integration of social cues. 


Amygdalo-hippocampal circuits mediating the integration of vomeronasal information during approach behaviours

Human reproductive behaviour manifests complex psychological exposition. The cognitive mechanism underlying the psychological aspects of mate selection demonstrates multifaceted objectives and is highly influenced by multiple societal factors. We hypothesize that, huge amount of erogenous provoking social contents (primarily digital) are creating complex database in young human mindset which eventually creates confusion about mate selection, thus demonstrating erratic behavioural sequences. The orthodox reproductive necessities in humans have notably incorporated ‘pleasure’ preferences also. Our analysis denotes that these external factors are capable of manipulating the fundamental evolutionary aspects of reproductive protocol and provide various dimensions about courtship perspectives. We conceptualize the recent paradigms of human mate selection perspectives and predict the onset of ‘artificial’ mating behaviour in the young population, with emphasis on social dimension. According to our findings, we are focused on confusion, which may be crucial in causing irregular behavioural manifestations during partner selection.

Erratic behaviour of human mate selection; a manipulative nature of young mindset creating confusion

Larval recruitment is a key life history trait for fish, conditioning species’ fate at an individual, population and community level. This process has many involvements, from the connectivity of the population, stock replenishment or adult fitness, being very sensitive to environmental change and anthropogenic activities, including light pollution. The global extent of artificial light at night (ALAN) on marine organisms is twofold. In the first place, ALAN is ubiquitous, impacting a quarter of the world’s coastline and pervasive until 10 meters deep for 2.7 % of the world’s exclusive economic zone. In the second place, ALAN is omnipotent, impacting most of the marine taxa examined to date and influencing a wide range of biological and ecological processes. Nevertheless, light pollution remains unstudied for larval recruitment.
In the present work, we reproduced environmental light pollution in the wild, in the lagoon of Moorea, French Polynesia, to examine the impact on fish recruitment. We particularly focused on two of the most dominant species of coral reef damselfish: the yellowtail dascyllus (Dascyllus flavicaudus) and the blue-green chromis (Chromis viridis). Using multidisciplinary approach (such as otoliths extraction, nighttime behavioural test or respirometry test), we found out that light pollution increases the recruitment of fish larvae to corals exposed to light pollution, but has also drastic carry over effect on growth, metabolic rate, and survival. In other words, ALAN has the potential to attract organisms to a less suitable environment, generating a peculiar anthropogenic stressor.


The light pollution paradox: more fish in less sustainable habitat

Climate change has revived interest in context-dependent species interactions as abiotic environments shift rapidly. An integrative approach, focusing on physiological mechanisms influencing environmental tolerance and performance, provides valuable insights into this research. Using the invasive mosquitofish (Gambusia holbrooki) and the endangered Spanish toothcarp (Aphanius iberus) as a case study, we explored salinity impact on metabolism and behaviour. While it is well-documented that salinity influences mosquitofish capacity, the metabolic mechanisms and their impact on behaviour remain unexplored. Our goal is to examine metabolic and behavioural responses in different salinity conditions (freshwater and 15 psu) and the correlation between these variables. Using intermittent-flow respirometry, we measured standard metabolic rates (SMR), maximum metabolic rates (MMR), and absolute aerobic scope (AAS). Behavioural variables, like the proportion of time out of cover, were derived from speed, acceleration, and heading data using Raspberry Pi cameras. Our findings show that, with rising salinity, mosquitofish experience decreased metabolic rates, while toothcarp's rates remain stable, suggesting lower salinity tolerance in mosquitofish. In terms of behaviour, mosquitofish exhibit higher activity and trajectory count, while toothcarps cover a larger area with increased speed variability. Correlations between metabolism and behaviour are species-specific, with toothcarps' MMR significantly correlating with overall behaviour, particularly time out of cover, and mosquitofish's SMR correlating with the total distance moved. These insights highlight challenges in mosquitofish metabolism with higher salinities compared to endemic species. Furthermore, it emphasises the importance of considering intricate physiological mechanisms and their interactions with behaviour in the context-dependent competition of this globally expanding species.

Energetic Tango: Metabolic and Behavioural Interplay in the Competition Dynamics of Native and Invasive Freshwater Fish Amid Changing Salinities

The red rock shrimp, Lysmata californica, is a key species in Southern California's shallow coastal ecosystems where it faces environmental challenges due to seasonal and long-term changes in ocean pH and temperature. These environmental changes could potentially affect the shrimp's molting process by extending postmolt recovery and affecting mortality rates through altering behaviors. This study investigates the impacts of ocean acidification (OA) and warming (OW) conditions on key behaviors of L. californica during postmolt. Shrimp were exposed to a combination of pH (8.1 and 7.7) and temperature (12°C and 15°C) conditions for 50 days, during which we assessed feeding, cleaning behavior, and startle responses at two critical times postmolt (<15 hours and 5 days post-ecdysis). Our findings reveal that reduced pH significantly delays initiation of feeding postmolt and diminishes food consumption postmolt, while warmer temperature increases overall food consumption. Additionally, reduced pH conditions deterred cleaning behavior, whereas warmer temperature enhanced it. There were no discernible effects on startle responses across treatments postmolt. These results demonstrate that OA and OW conditions can alter important behaviors of L. californica during the critical postmolt period, temporarily disrupting their roles within mutualistic symbioses under ongoing climate change.

Eco-physiological Impacts of Ocean Acidification and Warming on Postmolt Red Rock Shrimp, Lysmata californica

Controlling on their body size, several bird taxa show cognitive abilities outperforming the mammals. This applies namely to parrots and passerines. Yet, in birds we still have only limited information about the physiological circuits regulating the nervous system including neuro-immune interactions. In our previous research we have identified a gene loss of an important neuro-immune regulator CNR2 that is specific for parrots. Compared to songbirds parrots show significantly higher expression of pro-inflammatory markers (IL1B and IL6) in brains of individuals with sterile-induced abdominal inflammation. Parrots frequently suffer from behavioural disorders (depression and feather plucking) that can result from impaired inflammatory regulation. Here we measured gene expression of important neuropeptides that are involved in regulation of behaviour in brains and periphery (ileum) of parrots during acute neuroinflammation. Unlike in brain in the ileum we have identified differential gene expression of POMC. Next we compared the patterns of neuropeptides expression during acute inflammation to chronic inflammation. We have revealed association between expression of key pro-inflammatory cytokines and several neuropeptides (NPY, PDYN, POMC, TAC1 and VIP). Finally, we followed the relationships between cytokine and neuropeptide expression and behavioural patterns in the experimental animals. Our results support the importance of neuro-immune interactions in regulation of avian behaviour during inflammatory conditions. Targeting neuropeptide expression during chronic inflammation could offer novel therapeutic strategies to treat depression-like disorders in parrots.

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Highly maintained heat production capacity contributes to the initial step of endothermal development in Pacific bluefin tuna

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