Stickleback Gasterosteus Aculeatus Research Articles

Do you remember? Within-generation and transgenerational heat stress memory of recurring marine heatwaves in threespine stickleback.

Marine heatwaves can have major and lasting effects on organism physiology and species persistence. Such temperature extremes are increasing in frequency, with consecutive heatwave events already occurring within the lifetime of many organisms. Heat stress memory (thermal priming) by individuals is a potential within-generation response to cope with recurring marine heatwaves. However, whether this form of biological memory can be inherited across generations is not well known. We used a three-generation experiment to investigate individual and transgenerational effects of single and recurring marine heatwaves on fitness-related traits using stickleback (Gasterosteus aculeatus) as a model species. We exposed adults (both sexes) to heatwaves and assessed female reproductive output in both the parent and offspring generation, and offspring (both sexes) survival, growth and behaviour to establish a holistic picture of potential heatwave effects on ectothermic fish. Exposure to single, extreme heatwaves lowered reproductive output, decreased offspring exploratory behaviour, impeded capacity to respond to further thermal stress and reduced long-term survival. However, prior experience of heatwaves (heat stress memory) mitigated some of these effects at both an individual (growth) and transgenerational (fecundity) level, indicating that species experiencing increasing heatwave frequency as part of ongoing climate change may cope better than previously thought.

Co-profiling of single-cell gene expression and chromatin landscapes in stickleback pituitary

The pituitary gland is a key endocrine gland with various physiological functions including metabolism, growth, and reproduction. It comprises several distinct cell populations that release multiple polypeptide hormones. Although the major endocrine cell types are conserved across taxa, the regulatory mechanisms of gene expression and chromatin organization in specific cell types remain poorly understood. Here, we performed simultaneous profiling of the transcriptome and chromatin landscapes in the pituitary cells of the three-spined stickleback (Gasterosteus aculeatus), which represents a good model for investigating the genetic mechanisms underlying adaptive evolution. We obtained pairwise gene expression and chromatin profiles for 5184 cells under short- and long-day conditions. Using three independent clustering analyses, we identified 16 distinct cell clusters and validated their consistency. These results advance our understanding of the regulatory dynamics in the pituitary gland and provide a reference for future research on comparative physiology and evolutionary biology.

Paternal response to novel predator exposure correlates with transgenerational response in offspring of threespined stickleback.

Parental experiences can alter offspring phenotypes via transgenerational plasticity (TGP), which may prime offspring to adaptively respond to novel stressors, including novel predators. However, we know little about the types of sensory cues (e.g. visual, olfactory) that parents use to recognize novel predators and the consequences for offspring. Individuals may respond to novel cues if they mimic historical cues or they may need multiple sensory cues to recognize and respond to novel stimuli. We exposed threespined stickleback (Gasterosteus aculeatus) males to a full factorial of visual and olfactory cues of a novel trout predator prior to fertilization and tested offspring for antipredator behaviour and survival against a live predator. Fathers exposed to visual cues oriented more to and spent time closer to the novel predator post-exposure on the first day. Paternal response to visual cues was echoed in their offspring: offspring of fathers exposed to visual cues were caught faster by a live predator, suggesting that multiple cues are not needed to induce a transgenerational effect. While visual cues elicited responses both within- and transgenerationally, they do not seem to result in adaptive priming in offspring, suggesting the possibility of maladaptive TGP in response to novel cues of predation risk.

Non-random mating behaviour between diverging littoral and pelagic three-spined sticklebacks in an invasive population from Upper Lake Constance.

Adaptive divergence and increased genetic differentiation among populations can lead to reproductive isolation. In Lake Constance, Germany, a population of invasive three-spined stickleback (Gasterosteus aculeatus) is currently diverging into littoral and pelagic ecotypes, which both nest in the littoral zone. We hypothesized that assortative mating behaviour contributes to reproductive isolation between these ecotypes and performed a behavioural experiment in which females could choose between two nest-guarding males. Behaviour was recorded, and data on traits relevant to mate choice were collected. Both females of the same and different ecotypes were courted with equal vigour. However, there was a significant interaction effect of male and female ecotypes on the level of aggression in females. Littoral females were more aggressive towards pelagic males, and pelagic females were more aggressive towards littoral males. This indicates rejection of males of different ecotypes in spite of the fact that littoral males were larger, more intensely red-coloured and more aggressive than the pelagic males-all mating traits female sticklebacks generally select for. This study documents the emergence of behavioural barriers during early divergence in an invasive and rapidly diversifying stickleback population and discusses their putative role in facilitating reproductive isolation and adaptive radiation within this species.

Quantifying animal social behaviour with ecological field methods.

Field studies of social behaviour are challenging due to the need to record or infer interactions between multiple individuals, often under suboptimal environmental conditions or with potential disturbance by observers. Due to the limited field techniques available, we present a novel method to quantify social behaviours in the field by comparing the counts of individuals caught in traps across multiple locations sampled simultaneously. The distribution of individuals between traps gives the extent of aggregation, and phenotypic data allow for inference of non-random assortment. As a case study, we applied this method to populations of three-spined sticklebacks (Gasterosteus aculeatus) in freshwater ponds, using minnow traps. As expected, we observed a strong trend for aggregation. We were able to describe the ecological drivers of aggregation, comparing environmental and phenotypic conditions across sites. Aggregation was not related to environmental parameters, but was negatively associated with the proportion of breeding males caught during the breeding season. No evidence for phenotypic assortment based on body size was found. These results demonstrate that widely available ecological equipment can address questions related to social behaviour. This cost-effective approach, avoiding the tagging of individuals and minimizing extended observer disturbance, can be applied across various habitats and species.

Reduced predation and competition from herring may have contributed to the increase of three-spined stickleback in the Baltic Sea

Abstract In many areas of the Baltic Sea, three-spined stickleback (Gasterosteus aculeatus) has increased several fold since the early 2000s. Two major planktivores of the Baltic Sea, Atlantic herring (Clupea harengus) and European sprat (Sprattus sprattus), may interact with stickleback via competition for food and predation, but the potential implications at the population level are still unknown. Here, we explored interactions between stickleback, herring, and sprat using (i) herring diet analyses and (ii) fish biomass estimates from hydroacoustics for 2001–2019 covering the largest Baltic Sea basins—the Central Baltic and the Bothnian Sea. The diet analyses revealed that stickleback is an important prey for large herring along the Swedish Baltic Sea coast, comprising up to 64% of the diet of individuals >22 cm. We found a negative influence of large herring (>18 cm) on stickleback in the Central Baltic, and a negative relationship between sprat and small herring (<18 cm) (pooled) and stickleback in the Bothnian Sea. The decline in herring and sprat population after the mid-1990s could have contributed to the increase in stickleback population via reduced predation and competition. Overall, herring may be an underappreciated piscivore, and high fishing pressure on herring could generate cascading effects on lower trophic levels.

Managing the tradeoff between reproduction and survival requires flexibility in behaviour and gene regulation in three-spined stickleback.

Predators exert a powerful selective force, however, predator avoidance can conflict with other important activities such as attracting mates. Decisions over whether to court mates versus avoiding predators are vital to fitness, yet the mechanistic underpinnings of how animals manage such tradeoffs are poorly understood. Here, we investigate the flexibility of behaviour and gene regulation in response to a tradeoff between avoiding predators (survival) and courting potential mates (reproduction) in three-spined stickleback (Gasterosteus aculeatus). We compared behavioural and transcriptomic responses of male sticklebacks faced with a courtship opportunity and cues of a predator simultaneously with the responses of males faced with a courtship opportunity or cues of a predator alone, and found that males behaviourally compromised courtship in favour of predator avoidance when faced with a tradeoff between them. The need to manage this tradeoff elicited dynamic changes in brain gene expression, and sets of functionally connected genes were organized into discrete modules based on co-expression. Additionally, we found that behavioural flexibility in response to tradeoffs corresponded to flexibility in gene regulatory network structure. Combined, these results uncover the coordinated response by the brain to a fundamental ecological tradeoff, providing insight into the structure and function of genetic networks underpinning how animals make fitness-influencing decisions.

Predicting population-level impacts of projected climate heating on a temperate freshwater fish.

Climate heating has the potential to drive changes in ecosystems at multiple levels of biological organization. Temperature directly affects the inherent physiology of plants and animals, resulting in changes in rates of photosynthesis and respiration, and trophic interactions. Predicting temperature-dependent changes in physiological and trophic processes, however, is challenging because environmental conditions and ecosystem structure vary across biogeographical regions of the globe. To realistically predict the effects of projected climate heating on wildlife populations, mechanistic tools are required to incorporate the inherent physiological effects of temperature changes, as well as the associated effects on food availability within and across comparable ecosystems. Here we applied an agent-based bioenergetics model to explore the combined effects of projected temperature increases for 2100 (1.4, 2.7, and 4.4°C), and associated changes in prey availability, on three-spined stickleback (Gasterosteus aculeatus) populations representing latitudes 50, 55, and 60°N. Our results showed a decline in population density after a simulated 1.4°C temperature increase at 50°N. In all other modeled scenarios there was an increase (inflation) in population density and biomass (per unit area) with climate heating, and this inflation increased with increasing latitude. We conclude that agent-based bioenergetics models are valuable tools in discerning the impacts of climate change on wild fish populations, which play important roles in aquatic food webs.

Beyond the mosaic model of brain evolution: Rearing environment defines local and global plasticity.

Comparative animal studies have identified a trend toward a more global structural organization as brains become larger, suggesting that brain regions grow in sync as predicted by the concerted model of brain evolution. At the same time, brain plasticity studies have identified a boost in local brain structure triggered by the environment, suggesting that brain regions grow independently, as predicted by the mosaic model. Nevertheless, it is unclear whether the environment can also trigger shifts toward a more global brain structure, that is, whether phenotypic plasticity proceeds in a concerted fashion. Here, we examined the impact of radically different rearing environments on brain organization in a teleost fish, the three-spined stickleback (Gasterosteus aculeatus). We computed novel indices of local and global brain structure across groups reared in the two environments and entered them as predictors of differences in brain and body sizes. Changes in local brain structure predicted differences in both body and brain sizes, whereas changes in global brain structure only predicted differences in brain size. Our findings highlight the emergence of brain plasticity in a population as local and global changes that are both compatible with the concerted model.

A reinvestigation of cognitive styles in sticklebacks: decision success varies with behavioral type.

The "cognitive styles" hypothesis suggests that individual differences in behavior are associated with variation in cognitive performance via underlying speed-accuracy trade-offs. While this is supported, in part, by a growing body of evidence, some studies did not find the expected relationships between behavioral type and cognitive performance. In some cases, this may reflect methodological limitations rather than the absence of a true relationship. The physical design of the testing arena and the number of choices offered in an assay can hinder our ability to detect inter-individual differences in cognitive performance. Here, we re-investigated the cognitive styles hypothesis in threespine stickleback (Gasterosteus aculeatus), adapting the maze design of a previous study which found no cost to decision success by faster (bolder) individuals. We used a similar design but increased the size of the maze and incorporated an additional choice in the form of a third maze arm. We found, in accordance with cognitive style expectations, that individuals who were consistently slower to emerge from the start chamber made fewer errors than fish that emerged faster. Activity in an open field test, however, did not show evidence of a relationship with decision success, possibly due to the low number of repeated observations per fish in this separate assay. Our results provide further empirical support for the cognitive styles hypothesis and highlight important methodological aspects to consider in studies of inter-individual differences in cognition.

Prickly postglacial pioneers: freshwater plankton community composition influences fatty acid desaturase (FADS2) copy number in southern Greenland threespine sticklebacks

The adaptation of marine fish to freshwater environments includes prodigious examples of rapid evolution.Given the scarcity of ω‐3 long‐chain polyunsaturated fatty acids (LC‐PUFA) in freshwater, we expect selection to be strong on fish transitioning to freshwater habitats and yet the underlying ecological causes of genomic and phenotypic differentiation are poorly understood for traits associated with lipid content and composition. Threespine stickleback Gasterosteus aculeatus have repeatedly colonized, and adapted to, freshwater habitats across the Northern Hemisphere. These freshwater populations often show elevated copy number of the fatty acid desaturase 2 gene (FADS2), which increases the biosynthetic capacity of LC‐PUFA. The starkly lower content of LC‐PUFA in freshwater compared to marine prey, especially docosahexaenoic acid (DHA), likely imposes strong positive selection on freshwater fish for either increased biosynthesis or greater dietary acquisition of LC‐PUFA. The recently colonized and relatively undisturbed threespine stickleback populations in postglacial coastal lakes of southern Greenland offer an exceptional opportunity to study how variation in the copy number of FADS2 is related to abiotic and biotic conditions of lakes and their morphometry. As expected, given its position on the stickleback X chromosome, we found strong sexual dimorphism in FADS2 copy number in all populations (19 freshwater, 1 marine and 1 brackish), and an increased dimorphism in some freshwater populations. We also found that FADS2 copy number was negatively correlated, for both males and females, with the abundance of copepods, which are a DHA‐rich food source in the zooplankton community. Overall, our results suggest that the prey community context of lakes might influence the process of metabolic adaptation of marine fish colonizing freshwater ecosystems.

Genomic Sequence of the Threespine Stickleback Iridovirus (TSIV) from Wild Gasterosteus aculeatus in Stormy Lake, Alaska.

The threespine stickleback iridovirus (TSIV), a double-stranded DNA virus, was the first megalocytivirus detected in wild North American fishes. We report a second occurrence of TSIV in threespine stickleback (Gasterosteus aculeatus) from Stormy Lake, Alaska, and assemble a nearly complete genome of TSIV. The 115-kilobase TSIV genome contains 94 open reading frames (ORFs), with 91 that share homology with other known iridoviruses. We identify three ORFs that likely originate from recent lateral gene transfers from a eukaryotic host and one ORF with homology to B22 poxvirus proteins that likely originated from a lateral gene transfer between viruses. Phylogenetic analysis of 24 iridovirus core genes and pairwise sequence identity analysis support TSIV as a divergent sister taxon to other megalocytiviruses and a candidate for a novel species designation. Screening of stickleback collected from Stormy Lake before and after a 2012 rotenone treatment to eliminate invasive fish shows 100% positivity for TSIV in the two years before treatment (95% confidence interval: 89-100% prevalence) and 0% positivity for TSIV in 2024 after treatment (95% confidence interval: 0 to 3.7% prevalence), suggesting that the rotenone treatment and subsequent crash and reestablishment of the stickleback population is associated with loss of TSIV.

Mitochondrial functions and fatty acid profiles in fish heart: an insight into physiological limitations linked to thermal tolerance and age.

Heart failure is among the first major consequences of heat stress in aquatic ectotherms. Mitochondria produce most of the ATP used by the heart and represent almost half of the volume in cardiac cells. It has therefore been hypothesized that mitochondrial dysfunction may be a major cause of heart failure associated with heat stress. The present study aims to investigate if CTmax is linked to the thermal sensitivity of cardiac mitochondria in the three-spined stickleback (Gasterosteus aculeatus), and if it is influenced by heart fatty acid composition and age. To do so, we measured the CTmax of 30 fish. The cardiac mitochondrial oxygen consumption was measured by high resolution respirometry at three temperatures and heart lipid profiles were obtained by gas chromatography (GC) coupled with a flame ionization detector (FID). Fish age was estimated via otolith readings. Fatty acid profiles showed no correlation with CTmax, but EPA levels were higher in older individuals. Mitochondrial respiration was measured in 35 fish using high-resolution respirometry. It was strongly affected by temperature and showed a drastic drop in OXPHOS respiration fed by complex I and complex I+complex II, while uncoupled respiration plateaued at CTmax temperature. Our results suggest that complex I is an important modulator of the impact of temperature on mitochondrial respiration at high temperatures but is not the main limiting factor in physiological conditions (maximal OXPHOS). Mitochondrial respiration was also affected by fish age, showing a general decrease in older individuals.

Tapeworm infection affects sleep-like behavior in three-spined sticklebacks

Sleep is a complex and conserved biological process that affects several body functions and behaviors. Evidence suggests that there is a reciprocal interaction between sleep and immunity. For instance, fragmented sleep can increase the probability of parasitic infections and reduce the ability to fight infections. Moreover, viral and bacterial infections alter the sleep patterns of infected individuals. However, the effects of macro-parasitic infections on sleep remain largely unknown, and measuring sleep in non-model organisms remains challenging. In this study, we investigated whether macro-parasite infections could alter sleep-like behavior of their hosts. We experimentally infected three-spined sticklebacks (Gasterosteus aculeatus), a freshwater fish, with the tapeworm Schistocephalus solidus and used a hidden Markov model to characterize sleep-like behavior in sticklebacks. One to four days after parasite exposure, infected fish showed no difference in sleep-like behavior compared with non-exposed fish, and fish that were exposed-but-not-infected only showed a slight reduction in sleep-like behavior during daytime. Twenty-nine to 32 days after exposure, infected fish showed more sleep-like behavior than control fish, while exposed-but-not-infected fish showed overall less sleep-like behavior. Using brain transcriptomics, we identified immune- and sleep-associated genes that potentially underlie the observed behavioral changes. These results provide insights into the complex association between macro-parasite infection, immunity, and sleep in fish and may thus contribute to a better understanding of reciprocal interactions between sleep and immunity.

Osmoregulation and reproduction: evolutionary trends in prolactin functions from fish to mammals

The study of prolactin function evolution provides key insights into the diverse effects of this hormone in mammals, both in health and disease, which is relevant from both theoretical and practical perspectives. This article reviews both original and literature data concerning the role of prolactin and its receptors in regulating the sexual dimorphism of freshwater adaptation in the three-spined stickleback Gasterosteus aculeatus L. It is demonstrated that mRNA expression of prolactin gene 1 (one of two prolactin paralogs) and its receptor PRLRA increases in the brains of female sticklebacks only upon transitioning to freshwater. The brain and kidneys of sticklebacks, as androgen-dependent organs, exhibit sex-dependent expression of Prlra in seawater. It is suggested that sex-dependent osmoregulatory effects of prolactin are mediated through the PRLRA receptor in these organs. The PRLRB receptor, expressed in the kidneys and brains of sticklebacks regardless of sex in seawater, shows increased sensitivity to reduced salinity, suggesting a more active role in implementing sex-independent osmoregulatory functions of prolactin. Gills and intestines, as osmoregulatory organs, express the PRLRA and PRLRB receptors independent of sex in both seawater and freshwater. With freshwater adaptation, there is a concurrent increase in the expression of Prl1 in the brains of females and the expression of Atp1a1 (α1a subunit of Na+/K+-ATPase), Nhe3 (NHE3 sodium-proton antiport gene), and Ecac (epithelial calcium channel gene) in their gills. It is presumed that these gill genes are under positive control by prolactin. Exploring the potential for prolactin’s osmoregulatory function in mammals revealed that it may manifest in conditions such as pathologies accompanied by increased expression of prolactin receptor isoforms in osmoregulatory organs. One of such pathologies is cholestasis in female rats, which was associated with an increase in Prlr isoform expression and changes in activity and ratio of Na+/K+-ATPase subunits in the kidney. Thus, it is concluded that in fish, the osmoregulatory function of prolactin is sex-dependent, while in mammals, it may manifest under conditions of disrupted water-salt exchange.

Effects of temporal hydrologic shifts on the population biology of an endangered freshwater fish in a dryland river ecosystem

AbstractSpecies occupying dryland river ecosystems often experience “boom‐and‐bust” demographic cycles that coincide with shifts in habitat availability. Knowing whether declines are within natural thresholds versus those caused by acute human disturbance is critical for managing protected species. We investigated temporal shifts in abundance and habitat use of an endangered population of the threespine stickleback Gasterosteus aculeatus in southern California, where a Mediterranean climate leads to ephemeral habitat in one of the regions' least hydrologically modified rivers, the Santa Clara River. We conducted population surveys over a period of below‐average rainfall in the upper watershed in Soledad Canyon, with predefined reaches surveyed multiple times per year to capture different hydrologic conditions. Abundances were stable across years but varied significantly depending on location, with some reaches remaining dry and others drying seasonally to varying degrees. Occupancy models showed that the presence of stable perennial reaches, drying regime, and other site‐specific factors were important predictors of habitat use, and that certain reaches may be key to ensuring source‐sink dynamics as flow dissipates over the dry season. Low occupancy in two sections was driven by different predominant mechanisms, one by diel cycles of evapotranspiration and the other by cattails (Typha spp.), with both having greater effects during the hotter, drier parts of the year. As dryland river ecosystems are vulnerable to the effects of anthropogenic‐induced climate change, this study demonstrates how temporal monitoring can delimit dry‐state benchmarks for improving management interventions (i.e., translocation and habitat restoration) for protected species under conditions that are predicted to worsen in the coming years.

Changes in gill neuroepithelial cells and morphology of threespine stickleback (Gasterosteus aculeatus) to hypoxia and simulated ocean acidification.

Coastal marine environments are characterized by daily, seasonal and long-term changes in both O2 and CO2, driven by local biotic and abiotic factors. The neuroepithelial cells (NECs) of fish are thought to be the putative chemoreceptors for sensing oxygen and CO2, and, thus, NECs play a key role in detecting these environmental changes. However, the role of NECs as chemosensors in marine fish remains largely understudied. In this study, the NECs of marine threespine sticklebacks (Gasterosteus aculeatus) were characterized using immunohistochemistry. We then determined if there were changes in NEC size and density, and in gill morphology in response to either mild (10kPa) or moderate (6.8kPa) hypoxia and two levels of elevated CO2 (1,500 and 3,000 µatm). We found that the NECs of stickleback contained synaptic vesicles and were innervated, and were 50-300% larger and 2 to 4 times more abundant than in other similar sized freshwater fishes. NEC size and density were largely unaffected by exposure to hypoxia, but there was a 50% decrease in interlamellar cell mass (ILCM) in response to mild and moderate hypoxia. NECs increased in size, but not abundance in response to elevated CO2. Moreover, fish exposed to moderate or elevated CO2 had 53-78% larger ILCMs compared to control fish. Our results demonstrated that adult marine sticklebacks have NECs that can respond to environmentally relevant pCO2 and likely hypoxia, which highlights the importance of NECs in marine fishes under the heterogeneity of environmental conditions in coastal areas.

Heterogeneous genomic architecture of skeletal armour traits in sticklebacks.

Whether populations adapt to similar selection pressures using the same underlying genetic variants depends on population history and the distribution of standing genetic variation at the metapopulation level. Studies of sticklebacks provide a case in point: when colonising and adapting to freshwater habitats, three-spined sticklebacks (Gasterosteus aculeatus) with high gene flow tend to fix the same adaptive alleles in the same major loci, whereas nine-spined sticklebacks (Pungitius pungitius) with limited gene flow tend to utilize a more heterogeneous set of loci. In accordance with this, we report results of quantitative trait locus (QTL) analyses using a backcross design showing that lateral plate number variation in the western European nine-spined sticklebacks mapped to three moderate-effect QTL, contrary to the major-effect QTL in three-spined sticklebacks and different from the four QTL previously identified in the eastern European nine-spined sticklebacks. Furthermore, several QTL were identified associated with variation in lateral plate size, and three moderate-effect QTL with body size. Together, these findings indicate more heterogenous and polygenic genetic underpinnings of skeletal armour variation in nine-spined than three-spined sticklebacks, indicating limited genetic parallelism underlying armour trait evolution in the family Gasterostidae.

Alternative splicing in parallel evolution and the evolutionary potential in sticklebacks.

Repeatability of adaptation to similar environments provides opportunity to evaluate the predictability of natural selection. While many studies have investigated gene expression differences between populations adapted to contrasting environments, the role of post-transcriptional processes such as alternative splicing has rarely been evaluated in the context of parallel adaptation. To address the aforementioned knowledge gap, we reanalysed transcriptomic data from three pairs of threespine stickleback (Gasterosteus aculeatus) ecotypes adapted to marine or freshwater environment. First, we identified genes with repeated expression or splicing divergence across ecotype pairs, and compared the genetic architecture and biological processes between parallelly expressed and parallelly spliced loci. Second, we analysed the extent to which parallel adaptation was reflected at gene expression and alternative splicing levels. Finally, we tested how the two axes of transcriptional variation differed in their potential for evolutionary change. Although both repeated differential splicing and differential expression across ecotype pairs showed tendency for parallel divergence, the degree of parallelism was lower for splicing than expression. Furthermore, parallel divergences in splicing and expression were likely to be associated with distinct cis-regulatory genetic variants and functionally unique set of genes. Finally, we found that parallelly spliced genes showed higher nucleotide diversity than parallelly expressed genes, indicating splicing is less susceptible to genetic variation erosion during parallel adaptation. Our results provide novel insight into the role of splicing in parallel adaptation, and underscore the contribution of splicing to the evolutionary potential of wild populations under environmental change.

The transgenerational consequences of paternal social isolation and predation exposure in threespined sticklebacks.

Parents routinely encounter stress in the ecological environment that can affect offspring development (transgenerational plasticity: TGP); however, parents' interactions with conspecifics may alter how parents respond to ecological stressors. During social buffering, the presence of conspecifics can reduce the response to or increase the speed of recovery from a stressor. This may have cascading effects on offspring if conspecifics can mitigate parental responses to ecological stress in ways that blunt the transmission of stress-induced transgenerational effects. Here, we simultaneously manipulated both paternal social isolation and experience with predation risk prior to fertilisation in threespined stickleback (Gasterosteus aculeatus). We generated offspring via in-vitro fertilisation to allow us to isolate paternal effects mediated via sperm alone (i.e. in the absence of paternal care). If social buffering mitigates TGP induced by paternal exposure to predation risk, then we expect the transgenerational effects of predation exposure to be weaker when a conspecific is present compared to when the father is isolated. Offspring of predator-exposed fathers showed reduced anxiety-like behaviour and tended to be captured faster by the predator. Fathers who were socially isolated also had offspring that were captured faster by a live predator, suggesting that paternal social isolation may have maladaptive effects on how offspring respond to ecological stressors. Despite additive effects of paternal social isolation and paternal predation risk, we found no evidence of an interaction between these paternal treatments, suggesting that the presence of a conspecific did not buffer fathers and/or offspring from the effects of predation risk. Our results suggest that socially induced stress is an important, yet underappreciated, mediator of TGP and can elicit transgenerational effects even in species that do not form permanent social groups. Future studies should therefore consider how the parental social environment can affect both within and trans-generational responses to ecological stressors.