The effect of observing trained conspecifics on the performance and motivation of goldfish, Carassius auratus, in a spatial task.

Spatial and social cognition are two aspects of fish behaviour that have been subject to an increasing amount of research in recent years, but few have investigated potential behaviour overlaps. Testing the ability for an individual to socially learn a spatial task would bridge this gap in understanding. We provided naïve goldfish, Carassius auratus, the opportunity to observe a trained conspecific navigate a T-shaped maze, and then recorded how many trials it took for them to learn the maze, time taken per trial, motivation, and acceptance of the food reward. We also recorded how many trials it took a control group to learn the maze without the opportunity to observe a demonstrator. The observer group took significantly longer to learn the maze than the control group. Although the observer group were significantly less motivated (trials without a choice made), they were significantly more likely to accept the food reward. The social learning of reward acceptance was taking place, but the process of the demonstration disrupted the training of the spatial task, with possible explanations as the passenger effect and trade-off mechanism being discussed. Future studies are needed to determine whether goldfish can acquire spatial information socially; however, this study contributes to the feasibility of studying social learning of environmentally information in goldfish.

Visual shape discrimination in goldfish, modelled with the neural circuitry of optic tectum and torus longitudinalis.

There is no satisfactory neurally-based theory as to how vertebrates that lack a neocortex discriminate even simple geometric shapes. In fishes, an intact optic tectum is necessary for such discriminations, but physiological studies of it have found nothing like the hierarchically arranged feature detecting neurons of mammalian visual cortex. Here, a neural model attempts a solution by basing shape discrimination upon the responses of only those elementary detectors (e.g. of size) that are within a focus of attention, formed by a winner-take-all arrangement of retinotopically mapped units representing tectal pyramidal cells. While this relatively primitive mechanism could recognize an object irrespective of position in space, it fails to distinguish patterns that differ only in their features' spatial relationships. The model's solution - imitating goldfish that naturally attend to the top of shapes - is to shift attention to the edges of a shape by spatially offsetting inputs to the pyramidal neurons, effected by the torus longitudinalis and its prolific synapses on pyramidal dendrites. The model's shape discrimination was compared to an extensive behavioral study using shapes with points and projections. In one test series fish were sensitive to the relative number of points on the tops of shapes. In another, fish were trained to discriminate points on the sides. By using different offset connections and only one elementary feature detector for small dark spots, the model successfully emulated the two sets of goldfish data, as judged by significant correlations between model response and fish discrimination.

Alfaxalone does not have long-term effects on goldfish pyramidal neuron action potential properties or GABAA receptor currents.

Anesthetics have varying physiological effects, but most notably alter ion channel kinetics. Alfaxalone is a rapid induction and washout neuroactive anesthetic, which potentiates γ-aminobutyric acid (GABA)-activated GABAA receptor (GABAA-R) currents. This study aims to identify any long-term effects of alfaxalone sedation on pyramidal neuron action potential and GABAA-R properties, to determine if its impact on neuronal function can be reversed in a sufficiently short timeframe to allow for same-day electrophysiological studies in goldfish brain. The goldfish (Carassius auratus) is an anoxia-tolerant vertebrate and is a useful model to study anoxia tolerance mechanisms. The results show that alfaxalone sedation did not significantly impact action potential properties. Additionally, the acute application of alfaxalone onto naive brain slices caused the potentiation of whole-cell GABAA-R current decay time and area under the curve. Following whole-animal sedation with alfaxalone, a 3-h wash of brain slices in alfaxalone-free saline, with saline exchanged every 30 min, was required to remove any potentiating impact of alfaxalone on GABAA-R whole-cell currents. These results demonstrate that alfaxalone is an effective anesthetic for same-day electrophysiological experiments with goldfish brain slices.

High-density lipoprotein remodeling affects the osmotic properties of plasma in goldfish under critical salinity.

To investigate the stress response and physiological adaptations of goldfish (Carassius auratus) to critical salinity (CS) waters, we analyzed high-density lipoprotein (HDL) stoichiometry, stress markers (cortisol, glucose), and plasma osmotic properties (Na+ , osmolality, water content) using ichthyology, biochemistry, and proteomics approaches. After 21 days of exposure to CS, plasma concentrations of cortisol, glucose, and Na+ increased, indicating stress. Total plasma osmolality (Osmtotal ) and osmolality generated by inorganic (Osminorg ) and organic osmolytes (Osmorg ) also increased, the latter by ~2%. We associated the increase of Osmorg with (1) increased metabolite concentration (glucose), (2) dissociation of HDL particles resulting in increased HDL number per unit plasma volume (~1.5-2-fold) and (3) increased HDL osmotic activity. HDL remodeling may be the reason for the redistribution of bound and free water in plasma, which may contribute to water retention in plasma and, at the same time, to hemodynamic disturbances under CS conditions. The study's findings suggest that HDL remodeling is an important mechanism for maintaining osmotic homeostasis in fish, which is consistent with current capillary exchange models in vertebrates.

IoT and ML approach for ornamental fish behaviour analysis.

Ornamental fish keeping is the second most preferred hobby in the world and it provides a great opportunity for entrepreneurship development and income generation. Controlling the environment in ornamental fish farm is a considerable challenge because it is affected by a variety of parameters like water temperature, dissolved oxygen, pH, and disease occurrences. One particular interesting ornamental fish species is goldfish (Carassius auratus). Machine learning (ML) and deep learning technique have significant potential in analysing voluminous data collected from fish farm. Through this technique, the fish farmers can get insight on feeding behaviour, fish growth patterns, predict diseases/stress, and environmental factors affecting fish health. The aim of the study is to analyze the behavioural changes in goldfish due to alterations in environmental parameters (water temperature and dissolved oxygen). Decision tree, Naïve Bayes classifier, K-nearest neighbour (KNN), and linear discriminant analysis (LDA) were used to analyse the behavioural change data. To compare the performance between all four classifiers, cross validation and confusion matrix used. The cross-validation error of LDA, Naïve Bayes classification, KNN and decision tree was 19.86, 28.08, 30.14 and 13.78 respectively. Decision tree was proved to be the most accurate and effective classifier. Different temperature and DO range were taken to predict fish behaviour. Some findings are, the behaviour of fish was rest between temperature 37.85 °C and 40.535 °C, erratic when temperature was greater than or equal to 40.535 °C, gasping when temperature was between 37.85 and 40.535 °C and when DO concentration was less than 6.58 mg/L. Blood parameter analysis has been done to validate the change in external behaviours with change in physiological parameters.

POTASSIUM CHLORIDE ADMINISTERED VIA FOUR ROUTES FOR EUTHANASIA OF ANESTHETIZED GOLDFISH (CARASSIUS AURATUS).

Immersion overdose in tricaine methanesulfonate (MS-222) is ineffective for euthanasia of adult goldfish (Carassius auratus), and investigation of alternative techniques is warranted. This study evaluated potassium chloride (KCl) administered via four routes for goldfish euthanasia. Thirty clinically healthy adult goldfish (17.2-41.9 g) were anesthetized via immersion in buffered MS-222 (300 mg/L) for 5 min beyond anesthetic induction. Fish were then randomly administered one of five treatments: KCl (333 mg/mL, 4.5 mmol/mL, 4.5 mEq/mL) via intracardiac injection at 10 mEq/kg (IC), intracoelomic injection (ICe) or bilateral topical delivery over the gill filaments (T) at 100 mEq/kg, or 90-min immersion at 4,500 mEq/L (saturated solution) dissolved in induction solution (water [W]), or no treatment (control [C]). Following treatment, all fish were moved to anesthetic-free freshwater. Serial heart rates were collected via Doppler device until sound cessation or recovery. Median (range) time to perform treatments was 5.3 (1.2-8.0) min, 3 (3-3) s, and 10 (10-10) s in IC, ICe, and T, respectively. Doppler cessation occurred in 6/6, 6/6, 6/6, 6/6, and 0/6 fish in median (range) times of 3 (0-210), 18 (10-45), 118 (90-390), and 150 (60-240) min in IC, ICe, T, W, and C, respectively; this was significantly different between groups (P = 0.002). Following or during treatment, 1/6, 2/6, 6/6, and 4/6 fish in IC, ICe, T, and W, respectively, exhibited intermittent transient jerking movements. Median (range) time to recovery in C was 5.5 (3.5-6.5) min. All administration routes were effective, but time to Doppler cessation varied and transient movements were noted. Preliminary follow-up research determined that return to MS-222 induction solution following ICe KCl treatment abolished movements. ICe KCl at 100 mEq/kg was technically simple and resulted in Doppler sound cessation in a clinically relevant time frame in 6/6 anesthetized goldfish. Return to MS-222 immersion following KCl administration is recommended.

Nocturnal melatonin increases glucose uptake via insulin-independent action in the goldfish brain.

Melatonin, a neurohormone nocturnally produced by the pineal gland, is known to regulate the circadian rhythm. It has been recently reported that variants of melatonin receptors are associated with an increased risk of hyperglycemia and type 2 diabetes, suggesting that melatonin may be involved in the regulation of glucose homeostasis. Insulin is a key hormone that regulates circulating glucose levels and cellular metabolism after food intake in many tissues, including the brain. Although cells actively uptake glucose even during sleep and without food, little is known regarding the physiological effects of nocturnal melatonin on glucose homeostasis. Therefore, we presume the involvement of melatonin in the diurnal rhythm of glucose metabolism, independent of insulin action after food intake. In the present study, goldfish (Carassius auratus) was used as an animal model, since this species has no insulin-dependent glucose transporter type 4 (GLUT4). We found that in fasted individuals, plasma melatonin levels were significantly higher and insulin levels were significantly lower during the night. Furthermore, glucose uptake in the brain, liver, and muscle tissues also significantly increased at night. After intraperitoneal administration of melatonin, glucose uptake by the brain and liver showed significantly greater increases than in the control group. The administration of melatonin also significantly decreased plasma glucose levels in hyperglycemic goldfish, but failed to alter insulin mRNA expression in Brockmann body and plasma insulin levels. Using an insulin-free medium, we demonstrated that melatonin treatment increased glucose uptake in a dose-dependent manner in primary cell cultures of goldfish brain and liver cells. Moreover, the addition of a melatonin receptor antagonist decreased glucose uptake in hepatocytes, but not in brain cells. Next, treatment with N1-acetyl-5-methoxykynuramine (AMK), a melatonin metabolite in the brain, directly increased glucose uptake in cultured brain cells. Taken together, these findings suggest that melatonin is a possible circadian regulator of glucose homeostasis, whereas insulin acquires its effect on glucose metabolism following food intake.

Boundary vector cells in the goldfish central telencephalon encode spatial information.

Navigation is one of the most fundamental cognitive skills for the survival of fish, the largest vertebrate class, and almost all other animal classes. Space encoding in single neurons is a critical component of the neural basis of navigation. To study this fundamental cognitive component in fish, we recorded the activity of neurons in the central area of the goldfish telencephalon while the fish were freely navigating in a quasi-2D water tank embedded in a 3D environment. We found spatially modulated neurons with firing patterns that gradually decreased with the distance of the fish from a boundary in each cell's preferred direction, resembling the boundary vector cells found in the mammalian subiculum. Many of these cells exhibited beta rhythm oscillations. This type of spatial representation in fish brains is unique among space-encoding cells in vertebrates and provides insights into spatial cognition in this lineage.

Comparative Observation and Analysis of Preference Behavior Based on Three Types of Taxes and Locomotor Activity in the Goldfish, Carassius auratus.

Psychophysiological studies in vertebrates have focused on taxes as indicators of behavioral change. Actually, a considerable number of studies about anxiety-like and anti-anxiety-like behaviors involving geotaxis, scototaxis, and thigmotaxis have been conducted on fish. However, few analyses considering these behaviors based on taxes in fish have been conducted. Here, using goldfish, we measured the time spent in the bright or dark area of a horizontally long rectangular tank (HLRT), in the upper or lower area of a vertically long rectangular tank (VLRT), and in the central or edge area of a circular tank (CT), respectively, for the first 30 min and the last 30 min in a 3-h period after fish had been introduced to tanks. Dark, lower, and edge preference behaviors were observed for the first 30 min in all tanks. While dark and edge preference behaviors were maintained even for the last 30 min, the lower preference was lost. Swimming distance and the number of area crossings in each tank were also compared between the first 30 min and the last 30 min. Both decreased significantly or tended to decrease in the last 30 min in the HLRT and the CT, but no change was observed in the VLRT. These results suggest that, in goldfish, preference behavior is stable for a short time, and that environmental habituation may depend on the shape of the tank and the elapsed time.

Seasonal changes in membrane structure and excitability in retinal neurons of goldfish (Carassius auratus) under constant environmental conditions.

Seasonal modifications in the structure of cellular membranes occur as an adaptive measure to withstand exposure to prolonged environmental change. Little is known about whether such changes occur independently of external cues, such as photoperiod or temperature, or how they may impact the central nervous system. We compared membrane properties of neurons isolated from the retina of goldfish (Carassius auratus), an organism well adapted to extreme environmental change, during the summer and winter months. Goldfish were maintained in a facility under constant environmental conditions throughout the year. Analysis of whole-retina phospholipid composition using mass spectrometry-based lipidomics revealed a twofold increase in phosphatidylethanolamine species during the winter, suggesting an increase in cell membrane fluidity. Atomic force microscopy was used to produce localized, nanoscale-force deformation of neuronal membranes. Measurement of Young's modulus indicated increased membrane-cortical stiffness (or decreased elasticity) in neurons isolated during the winter. Voltage-clamp electrophysiology was used to assess physiological changes in neurons between seasons. Winter neurons displayed a hyperpolarized reversal potential (Vrev) and a significantly lower input resistance (Rin) compared with summer neurons. This was indicative of a decrease in membrane excitability during the winter. Subsequent measurement of intracellular Ca2+ activity using Fura-2 microspectrofluorometry confirmed a reduction in action potential activity, including duration and action potential profile, in neurons isolated during the winter. These studies demonstrate chemical and biophysical changes that occur in retinal neurons of goldfish throughout the year without exposure to seasonal cues, and suggest a novel mechanism of seasonal regulation of retinal activity.

Possible role of transcription factors (BSX, NKX2.1, IRX3 and SIRT1) in the regulation of appetite in goldfish (Carassius auratus).

The homeobox genes play important roles in the embryonic development of animals. Recent evidence suggests they might also regulate feeding and act as transcription factors of appetite regulators. Examples of these genes are a brain-specific homeobox transcription factor (BSX), NK2 homeobox 1 (NKX2.1) and the Iroquois homeobox 3 (IRX3). Sirtuin1 (SIRT1) acts as a transcription factor for nutrient (e.g. lipid, glucose) homeostasis and responds to stress and nutrient availability, and has been shown to interact with appetite regulators. Very little is known about the role of these genes in the regulation of feeding and nutrient homeostasis in fish. In this study, we assessed the roles of BSX, NKX2.1, IRX3 and SIRT1 in the central regulation of feeding in goldfish by examining their mRNA brain distribution, assessing the effects of fasting on their brain expression and assessing the effects of peripheral injections of cholecystokinin (CCK, a brain-gut peptide), on their brain expression. All genes showed a widespread distribution in the brain, with high levels in the hypothalamus. In both hypothalamus and telencephalon, fasting induced increases in BSX, IRX3 and NKX2.1 expressions but had no effect on SIRT1 expression levels. CCK injections increased hypothalamic expression levels of IRX3 and SIRT1, and telencephalic expression levels of NKX2.1 and SIRT1, with no effect on either hypothalamic BSX or NKX2.1 expression levels or telencephalon BSX or IRX3 expression levels. Our results suggest that, in goldfish as in mammals, central BSX, NKX2.1, IRX3 and SIRT1 are present in regions of the brain regulating feeding, are sensitive to nutrient status and interact with appetite-regulating peptides.

Evaluation of prolonged immersion in tricaine methanesulfonate for juvenile goldfish (Carassius auratus) euthanasia.

OBJECTIVE: Doses of buffered tricaine methanesulfonate (MS-222) up to 1000 mg/L for 15 minutes are reported inefficient to produce euthanasia in goldfish. The goal of this study was to determine if goldfish can be euthanized by more prolonged immersion in MS-222. ANIMALS: 24 healthy goldfish (weight range: 1 to 10 g) were randomly assigned to 4 groups of 6 fish. PROCEDURES: The first group (G1) was exposed to 500 mg/L buffered MS-222 for 15 minutes then placed in freshwater for 3 hours. The second (G2) and third groups (G3) were exposed to 1000 mg/L of buffered MS-222 for 15 minutes then placed in freshwater for 3 hours and 18 hours respectively. The fourth group (G4) was exposed to 1000 mg/L of buffered MS-222 for 60 minutes then placed in freshwater for 3 hours. Time to cessation and return of operculation were recorded. If the goldfish did not resume operculation, heart rate was evaluated by Doppler ultrasonic flow detector. RESULTS: Median times to apnea were 35 seconds at 1000 mg/L and 65 seconds at 500 mg/L. Re-operculation occurred only in G1 in 5 out of 6 individuals. All fish from G1, 3 fish from G2, 0 fish from G3, 1 fish from G4 had remaining heartbeats at the end of the observation period. CLINICAL RELEVANCE: Overall, a dose of 1000 mg/L of buffered MS-222 for 15 minutes was efficient to euthanize juvenile goldfish at 20 °C. Different fish body mass and water quality parameters might explain different results compared to previous studies.

Audiovisual integration in the Mauthner cell enhances escape probability and reduces response latency.

Fast and accurate threat detection is critical for animal survival. Reducing perceptual ambiguity by integrating multiple sources of sensory information can enhance perception and reduce response latency. However, studies addressing the link between behavioral correlates of multisensory integration and its underlying neural basis are rare. Fish that detect an urgent threat escape with an explosive behavior known as C-start. The C-start is driven by an identified neural circuit centered on the Mauthner cell, an identified neuron capable of triggering escapes in response to visual and auditory stimuli. Here we demonstrate that goldfish can integrate visual looms and brief auditory stimuli to increase C-start probability. This multisensory enhancement is inversely correlated to the salience of the stimuli, with weaker auditory cues producing a proportionally stronger multisensory effect. We also show that multisensory stimuli reduced C-start response latency, with most escapes locked to the presentation of the auditory cue. We make a direct link between behavioral data and its underlying neural mechanism by reproducing the behavioral data with an integrate-and-fire computational model of the Mauthner cell. This model of the Mauthner cell circuit suggests that excitatory inputs integrated at the soma are key elements in multisensory decision making during fast C-start escapes. This provides a simple but powerful mechanism to enhance threat detection and survival.

Interploidy gene flow involving the sexual-asexual cycle facilitates the diversification of gynogenetic triploid Carassius fish.

Asexual vertebrates are rare and at risk of extinction due to their restricted adaptability through the loss of genetic recombination. We explore the mechanisms behind the generation and maintenance of genetic diversity in triploid asexual (gynogenetic) Carassius auratus fish, which is widespread in East Asian fresh waters and exhibits one of the most extensive distribution among asexual vertebrates despite its dependence on host sperm. Our analyses of genetic composition using dozens of genetic markers and genome-wide transcriptome sequencing uncover admixed genetic composition of Japanese asexual triploid Carassius consisting of both the diverged Japanese and Eurasian alleles, suggesting the involvement of Eurasian lineages in its origin. However, coexisting sexual diploid relatives and asexual triploids in Japan show regional genetic similarity in both mitochondrial and nuclear markers. These results are attributed to a unique unidirectional gene flow from diploids to sympatric triploids, with the involvement of occasional sexual reproduction. Additionally, the asexual triploid shows a weaker population structure than the sexual diploid, and multiple triploid lineages coexist in most Japanese rivers. The generated diversity via repeated interploidy gene flow as well as an increased establishment of immigrants is assumed to offset the cost of asexual reproduction and might contribute to the successful broad distribution of this asexual vertebrate.

Expression of matrix metalloproteinases during axonal regeneration in the goldfish spinal cord.

Spinal cord injury in fish produces fibrous scar, but spontaneous axonal regeneration beyond the scar sometimes occurs. A previous study revealed that regenerating axons enter the scar through tubular structures with laminin, and that an increased number of axons within the tube is coincident with enlargement of the tube diameter and reduction of the fibrous scar area. The present study investigated the expression of matrix metalloproteinases (MMPs) that might play a role in the degradation of the extracellular matrix in fibrous scar tissue and in the remodeling of tubular structures. Spinal hemisection produced fibrous scar tissue in the lesion center, surrounded by nervous tissue. Two weeks after spinal lesioning, MMP-9 was expressed in some regenerating axons in the fibrous scar tissue. MMP-14 was expressed in the regenerating axons, as well as in glial processes in the fibrous scar tissue. MMP-2 was suggested to be expressed in mast cells in the fibrous scar. The mast cells were in contact with fibroblasts, and in close proximity to the basement membrane of tubular structures surrounding the regenerating axons. The present findings suggest that several MMPs are involved in axon regenerating processes following spinal cord injury in goldfish. MMP-9 and MMP-14 expressed in the regenerating axons might degrade extracellular matrix and support axonal growth deep into the fibrous scar tissue. MMP-14 expressed in glial cells and MMP-2 expressed in mast cells might also provide a beneficial environment for axonal regeneration, leading to successful motor recovery.

Expression of reproductive-related genes and changes in oocyte maturation of goldfish broodstock (Carassius auratus) following injection of different exogenous kisspeptins.

Kisspeptin, upstream of the hypothalamic-pituitary-gonadal axis, play an essential role in the reproductive process. In the present study, the effect of different types of kisspeptin, including goldfish (Carassius auratus) kiss1 kisspeptin (Kiss1), human kisspeptin (Hkiss) and their combination (Kiss1+H) on the reproductive-related genes (kiss1, Kissr and Cyp19) of adult female goldfish was investigated in comparison with Ovaprim (a synthetic GnRH hormone). Kiss1 and Hkiss were synthesized using a solid-phase synthesis approach. Peptides were injected at a dose of 100 µg/kg body weight. The brain and ovarian tissues of samples were separated for histological studies 24 hr post-injection. The expression of the kiss1, Kissr and Cyp19 genes was measured by RT-PCR. The results showed a significant increase in expression of the reproductive-related genes. Histological analysis revealed higher number of mature oocytes in kisspeptin treated groups compare to other ones. In conclusion, Hkiss and Kiss1+H are the most effective peptides in oocyte maturation and expression of reproductive-related genes. In addition, it seems that kisspeptins in other domestic animals can be used to stimulate the hypothalamus-pituitary-gonadal axis.

Fish swim water bulk displacement visualization with digital holographic interferometry.

A collimated transmission beam interferometer is used to measure the water motion provoked by the fish swimming through it. An indirect measurement of the fish motion impact in the water contained in a home-type aquarium is detected. Measurements of the whole aquarium are possible due to a large diameter collimated laser beam in the interferometer's object arm. This beam goes through the aquarium, and any perturbation inside it deflects the collimated beam. The interferometer detects a phase difference, i.e., the beam through the disturbed water undergoes different optical paths. This optical phase change was first demonstrated by means of a simple test using spherical steel marbles placed in a cuvette. For this, the small water movements for a single steel marble are detected with the acquired optical phase. Next, the aquarium optical phase results show water movements according to the fishes' size and swimming speed. It is worth mentioning that no additives were added to the aquarium's fresh water during the tests, so the water was crystal clear.

Hormonal Prostaglandin F2α Mediates Behavioral Responsiveness to a Species-Specific Multi-component Male Hormonal Sex Pheromone in a Female Fish.

Although hormonally-derived female sex pheromones have been well described in approximately a dozen species of teleost fish, only a few male sex pheromones have been characterized and the neuroendocrine underpinnings of behavioral responsiveness to them is not understood. Herein, we describe a study that addresses this question using the goldfish, Carassius auratus, an important model species of how hormones drive behavior in egg-laying teleost fishes. Our study had four components. First, we examined behavioral responsiveness of female goldfish and found that when injected with prostaglandin F2α (PGF2α), a treatment that drives female sexual receptivity, and found that they became strongly and uniquely attracted to the odor of conspecific mature males, while non-PGF2α-treated goldfish did not discern males from females. Next, we characterized the complexity and specificity of the male pheromone by examining the responsiveness of PGF2α-treated females to the odor of either mature male conspecifics or male common carp odor, as well as their nonpolar and polar fractions. We found that the odor of male goldfish was more attractive than that of male common carp, and that its activity was attributable to both its nonpolar and polar fractions with the later conveying information on species-identity. Third, we hypothesized that androstenedione (AD), a 19-carbon sex steroid produced by all male fish might be the nonpolar fraction and tested whether PGF2α-treated goldfish were attracted to either AD alone or as part of a mixture in conspecific water. We found that while AD was inactive on its own, it became highly attractive when added to previously unattractive female conspecific water. Lastly, in a test of whether nonhormonal conspecific odor might determine species-specificity, we added AD to water of three species of fish and found that while AD made goldfish water strongly attractive, its effects on other species holding water were small. We conclude that circulating PGF2α produced at the time of ovulation induces behavioral sensitivity to a male sex pheromone in female goldfish and that this male pheromone is comprised of AD and a mixture of body metabolites. Because PGF2α commonly mediates ovulation and female sexual behavior in egg-laying fishes, and AD is universally produced by male fishes as a precursor to testosterone, we suggest that these two hormones may have similar roles mediating male-female behavior and communication in many species of fish.

Bisphenols exert detrimental effects on neuronal signaling in mature vertebrate brains.

Bisphenols are important plasticizers currently in use and are released at rates of hundreds of tons each year into the biosphere1-3. However, for any bisphenol it is completely unknown if and how it affects the intact adult brain4-6, whose powerful homeostatic mechanisms could potentially compensate any effects bisphenols might have on isolated neurons. Here we analyzed the effects of one month of exposition to BPA or BPS on an identified neuron in the vertebrate brain, using intracellular in vivo recordings in the uniquely suited Mauthner neuron in goldfish. Our findings demonstrate an alarming and uncompensated in vivo impact of both BPA and BPS-at environmentally relevant concentrations-on essential communication functions of neurons in mature vertebrate brains and call for the rapid development of alternative plasticizers. The speed and resolution of the assay we present here could thereby be instrumental to accelerate the early testing phase of next-generation plasticizers.

Response of the thyroid axis and appetite-regulating peptides to fasting and overfeeding in goldfish (Carassius auratus).

The thyroid axis is a major regulator of metabolism and energy homeostasis in vertebrates. There is conclusive evidence in mammals for the involvement of the thyroid axis in the regulation of food intake, but in fish, this link is unclear. In order to assess the effects of nutritional status on the thyroid axis in goldfish, Carassius auratus, we examined brain and peripheral transcripts of genes associated with the thyroid axis [thyrotropin-releasing hormone (TRH), thyrotropin-releasing hormone receptors (TRH-R type 1 and 2), thyroid stimulating hormone beta (TSHß), deiodinase enzymes (DIO2, DIO3) and UDP-glucoronsyltransferase (UGT)] and appetite regulators [neuropeptide Y (NPY), proopiomelanocortin (POMC), agouti-related peptide (AgRP) and cholecystokinin (CCK)] in fasted and overfed fish for 7 and 14 day periods. We show that the thyroid axis responds to overfeeding, with an increase of brain TRH and TSHß mRNA expression after 14 days, suggesting that overfeeding might activate the thyroid axis. In fasted fish, hepatic DIO3 and UGT transcripts were downregulated from 7 to 14 days, suggesting a time-dependent inhibition of thyroid hormone degradation pathways. Nutritional status had no effect on circulating levels of thyroid hormone. Central appetite-regulating peptides exhibited temporal changes in mRNA expression, with decreased expression of the appetite-inhibiting peptide POMC from 7 to 14 days for both fasted and overfed fish, with no change in central NPY or AgRP, or intestinal CCK transcript expression. Compared to control fish, fasting increased AgRP mRNA expression at both 7 and 14 days, and POMC expression was higher than controls only at 7 days. Our results indicate that nutritional status time-dependently affects the thyroid axis and appetite regulators, although no clear correlation between thyroid physiology and appetite regulators could be established. Our study helps to fill a knowledge gap in current fish endocrinological research on the effects of energy balance on thyroid metabolism and function.