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Data associated with manuscript: "Foliar flavonoids across an elevation gradient: plasticity in response to UV, and links with floral pigmentation patterning"

There are two datasets provided in this excel file. The data are flavonoid abundances of Argentina anserina [=Potentilla anserina] exposed to two levels of ultraviolet light in a greenhouse. The "leaf only data" tab has the combined foliar abundances of the major classes of flavonoids detected: chalcone, flavonone, flavone, flavonol, flavon-3-ol, and proanthocyanidin. This dataset was used to analyze the effect of UV treatment and elevation on flavonoid production in leaves. The "leaf and petal matching cmpnds" is the abundance of the major flavonoid classes in leaves and petals for only the compounds that were detected in both tissue types. This dataset was used for all analyses that related leaf flavonoids with petal flavonoids and petal UV pigmentation.

Serial block-face scanning electron microscopy of adherent cells on thin plastic substrate – Data set 10

Serial block-face (SBF) scanning electron microscopy (SEM) is used for imaging the entire internal ultrastructure of cells, tissue samples or small organisms. We developed a workflow for SBF SEM of adherent cells, such as Giardia parasites and HeLa cells, attached to the surface of a plastic culture dish, which preserves the interface between cells and plastic substrate. Cells were embedded in situ on their substrate using silicone microwells and were mounted for cross-sectioning which allowed SBF imaging of large volumes and many cells. In total we provide 10 data sets with image series from SBF SEM of Giardia and HeLa cells prepared with protocol variants to improve the workflow. A detailed description of the methods and the data set is provided in the download container. Data set 10 comprises an image series of 255 images recorded of a HeLa cell adhered to the plastic substrate of a culture dish. SBF SEM was done at a section interval of 50 nm using the T1 detector of the SEM at high vacuum. Original pixel size was 10 nm. The data folder contains the raw image files, processed image files (see data set description for details of the processing), a video file of a processed image file series.

Serial block-face scanning electron microscopy of adherent cells on thin plastic substrate – Data set 05

Serial block-face (SBF) scanning electron microscopy (SEM) is used for imaging the entire internal ultrastructure of cells, tissue samples or small organisms. We developed a workflow for SBF SEM of adherent cells, such as Giardia parasites and HeLa cells, attached to the surface of a plastic culture dish, which preserves the interface between cells and plastic substrate. Cells were embedded in situ on their substrate using silicone microwells and were mounted for cross-sectioning which allowed SBF imaging of large volumes and many cells. In total we provide 10 data sets with image series from SBF SEM of Giardia and HeLa cells prepared with protocol variants to improve the workflow. A detailed description of the methods and the data set is provided in the download container. Data set 05 comprises an image series of 500 images recorded of a HeLa cell adhered to the plastic substrate of a culture dish. SBF SEM was done at a section interval of 50 nm using the DBS detector of the SEM at low vacuum (0.4 mbar). Original pixel size was 6 nm. The data folder contains the raw image files, processed image files (see data set description for details of the processing), a video file of a processed image file series.

Serial block-face scanning electron microscopy of adherent cells on thin plastic substrate – Data set 03

Serial block-face (SBF) scanning electron microscopy (SEM) is used for imaging the entire internal ultrastructure of cells, tissue samples or small organisms. We developed a workflow for SBF SEM of adherent cells, such as Giardia parasites and HeLa cells, attached to the surface of a plastic culture dish, which preserves the interface between cells and plastic substrate. Cells were embedded in situ on their substrate using silicone microwells and were mounted for cross-sectioning which allowed SBF imaging of large volumes and many cells. In total we provide 10 data sets with image series from SBF SEM of Giardia and HeLa cells prepared with protocol variants to improve the workflow. A detailed description of the methods and the data set is provided in the download container. Data set 03 comprises an image 3D model of a Giardia lamblia cell adhered to the plastic substrate of a culture dish. The model was generated by segmentation of the entire cell, the cell nuclei (red) and the ventral disc cytoskeleton (yellow) in an image series of 276 images which was recorded by SBF SEM (see dataset 01). Section interval was 50 nm and pixel size 10 nm. The data folder contains the model-file (Imaris-format) and a 360° rotation of the model as video file (mp4-format).

Serial block-face scanning electron microscopy of adherent cells on thin plastic substrate – Data set 02

Serial block-face (SBF) scanning electron microscopy (SEM) is used for imaging the entire internal ultrastructure of cells, tissue samples or small organisms. We developed a workflow for SBF SEM of adherent cells, such as Giardia parasites and HeLa cells, attached to the surface of a plastic culture dish, which preserves the interface between cells and plastic substrate. Cells were embedded in situ on their substrate using silicone microwells and were mounted for cross-sectioning which allowed SBF imaging of large volumes and many cells. In total we provide 10 data sets with image series from SBF SEM of Giardia and HeLa cells prepared with protocol variants to improve the workflow. A detailed description of the methods and the data set is provided in the download container. Data set 02 comprises an image series of 1462 images recorded of a Giardia lamblia cell adhered to the plastic substrate of a culture dish. SBF SEM was done at a section interval of 10 nm using the DBS detector of the SEM at low vacuum (0.5 mbar). Original pixel size was 5 nm. The data folder contains the raw image files, processed image files (see data set description for details of the processing), a video file of a processed image file series.

Serial block-face scanning electron microscopy of adherent cells on thin plastic substrate – Data set 09

Serial block-face (SBF) scanning electron microscopy (SEM) is used for imaging the entire internal ultrastructure of cells, tissue samples or small organisms. We developed a workflow for SBF SEM of adherent cells, such as Giardia parasites and HeLa cells, attached to the surface of a plastic culture dish, which preserves the interface between cells and plastic substrate. Cells were embedded in situ on their substrate using silicone microwells and were mounted for cross-sectioning which allowed SBF imaging of large volumes and many cells. In total we provide 10 data sets with image series from SBF SEM of Giardia and HeLa cells prepared with protocol variants to improve the workflow. A detailed description of the methods and the data set is provided in the download container. Data set 09 comprises an image series of 400 images recorded of a HeLa cell adhered to the plastic substrate of a culture dish. SBF SEM was done at a section interval of 10 nm using the DBS detector of the SEM at low vacuum (0.4 mbar). Original pixel size was 10 nm. The data folder contains the raw image files, processed image files (see data set description for details of the processing), a video file of a processed image file series.

Serial block-face scanning electron microscopy of adherent cells on thin plastic substrate – Data set 08

Serial block-face (SBF) scanning electron microscopy (SEM) is used for imaging the entire internal ultrastructure of cells, tissue samples or small organisms. We developed a workflow for SBF SEM of adherent cells, such as Giardia parasites and HeLa cells, attached to the surface of a plastic culture dish, which preserves the interface between cells and plastic substrate. Cells were embedded in situ on their substrate using silicone microwells and were mounted for cross-sectioning which allowed SBF imaging of large volumes and many cells. In total we provide 10 data sets with image series from SBF SEM of Giardia and HeLa cells prepared with protocol variants to improve the workflow. A detailed description of the methods and the data set is provided in the download container. Data set 08 comprises an image series of 299 images recorded of a HeLa cell adhered to the plastic substrate of a culture dish. SBF SEM was done at a section interval of 50 nm using the DBS detector of the SEM at low vacuum (0.4 mbar). Original pixel size was 3 nm. The data folder contains the raw image files, processed image files (see data set description for details of the processing), a video file of a processed image file series.

Serial block-face scanning electron microscopy of adherent cells on thin plastic substrate – Data set 04

Serial block-face (SBF) scanning electron microscopy (SEM) is used for imaging the entire internal ultrastructure of cells, tissue samples or small organisms. We developed a workflow for SBF SEM of adherent cells, such as Giardia parasites and HeLa cells, attached to the surface of a plastic culture dish, which preserves the interface between cells and plastic substrate. Cells were embedded in situ on their substrate using silicone microwells and were mounted for cross-sectioning which allowed SBF imaging of large volumes and many cells. In total we provide 10 data sets with image series from SBF SEM of Giardia and HeLa cells prepared with protocol variants to improve the workflow. A detailed description of the methods and the data set is provided in the download container. Data set 04 comprises an image series of 120 images recorded of a HeLa cell adhered to the plastic substrate of a culture dish. SBF SEM was done at a section interval of 50 nm using the T1 detector of the SEM at high vacuum. Original pixel size was 10 nm. The data folder contains the raw image files, processed image files (see data set description for details of the processing), a video file of a processed image file series.

Serial block-face scanning electron microscopy of adherent cells on thin plastic substrate – Data set 07

Serial block-face (SBF) scanning electron microscopy (SEM) is used for imaging the entire internal ultrastructure of cells, tissue samples or small organisms. We developed a workflow for SBF SEM of adherent cells, such as Giardia parasites and HeLa cells, attached to the surface of a plastic culture dish, which preserves the interface between cells and plastic substrate. Cells were embedded in situ on their substrate using silicone microwells and were mounted for cross-sectioning which allowed SBF imaging of large volumes and many cells. In total we provide 10 data sets with image series from SBF SEM of Giardia and HeLa cells prepared with protocol variants to improve the workflow. A detailed description of the methods and the data set is provided in the download container. Data set 07 comprises an image series of 318 images recorded of a HeLa cell adhered to the plastic substrate of a culture dish. SBF SEM was done at a section interval of 10 nm using the DBS detector of the SEM at low vacuum (0.4 mbar). Original pixel size was 8 nm. The data folder contains the raw image files, processed image files (see data set description for details of the processing), a video file of a processed image file series.

Serial block-face scanning electron microscopy of adherent cells on thin plastic substrate – Data set 06

Serial block-face (SBF) scanning electron microscopy (SEM) is used for imaging the entire internal ultrastructure of cells, tissue samples or small organisms. We developed a workflow for SBF SEM of adherent cells, such as Giardia parasites and HeLa cells, attached to the surface of a plastic culture dish, which preserves the interface between cells and plastic substrate. Cells were embedded in situ on their substrate using silicone microwells and were mounted for cross-sectioning which allowed SBF imaging of large volumes and many cells. In total we provide 10 data sets with image series from SBF SEM of Giardia and HeLa cells prepared with protocol variants to improve the workflow. A detailed description of the methods and the data set is provided in the download container. Data set 06 comprises an image series of 215 images recorded of a HeLa cell adhered to the plastic substrate of a culture dish. SBF SEM was done at a section interval of 10 nm using the DBS detector of the SEM at low vacuum (0.4 mbar). Original pixel size was 10 nm. The data folder contains the raw image files, processed image files (see data set description for details of the processing), a video file of a processed image file series.

Serial block-face scanning electron microscopy of adherent cells on thin plastic substrate – Data set 01

Serial block-face (SBF) scanning electron microscopy (SEM) is used for imaging the entire internal ultrastructure of cells, tissue samples or small organisms. We developed a workflow for SBF SEM of adherent cells, such as Giardia parasites and HeLa cells, attached to the surface of a plastic culture dish, which preserves the interface between cells and plastic substrate. Cells were embedded in situ on their substrate using silicone microwells and were mounted for cross-sectioning which allowed SBF imaging of large volumes and many cells. In total we provide 10 data sets with image series from SBF SEM of Giardia and HeLa cells prepared with protocol variants to improve the workflow. A detailed description of the methods and the data set is provided in the download container. Data set 01 comprises an image series of 276 images recorded of a Giardia lamblia cell adhered to the plastic substrate of a culture dish. SBF SEM was done at a section interval of 50 nm using the DBS detector of the SEM at low vacuum (0.5 mbar). Original pixel size was 4 nm. The data folder contains the raw image files, processed image files (see data set description for details of the processing), a video file of a processed image file series.

opencc-zeroMay 2024View details →

Finotypic plasticity: Predator-induced plasticity in fin size, darkness, and display behaviour in a teleost fish

Fish fins are remarkable devices of propulsion. Fin morphology is intimately linked to locomotor performance, and hence to behaviours that influence fitness, such as foraging and predator avoidance. This foreshadows a connection between fin morphology and variation in predation risk. Yet, whether prey can adjust fin morphology according to changes in perceived risk within their lifetime (a.k.a. predator-induced plasticity) remains elusive. Here, we quantify the structural size of five focal fins in crucian carp (Carassius carassius) following controlled manipulations to perceived predation risk (presence/absence of pike Esox lucius). We also assess if crucian carp respond to increased predation risk by shifts in dorsal fin colouration, and test for differences in how fish actively use their dorsal fins by quantifying the area of the fin displayed in behavioural trials. We find that crucian carp show phenotypic plasticity with regard to fin size as predator-exposed fish consistently have larger fins. Individuals exposed to perceived predation risk also increased dorsal fin darkness and actively displayed a larger area of the fin to potential predators. Our result thus provides compelling evidence for predator-induced fin enlargement, which should result in enhanced escape swimming performance. Moreover, fin-size plasticity may evolve synergistically with fin colouration and display behaviour, and we suggest that the adaptive value of this synergy is to enhance the silhouette of deep-bodied and hard-to-capture prey to deter gape-limited predators prior to an attack. Together, our results provide new perspectives on the role of predation risk for the development and evolution of fins.

opencc-zeroMay 2024View details →

Data from: Symbiont infection and psyllid haplotype influence phenotypic plasticity during host switching events

Many herbivorous insect species exhibit phenotypic plasticity when using multiple hosts, which facilitates survival in heterogeneous host environments. Physiological host acclimation is an important part of it, yet the effects of host acclimation on insect feeding behavior are not well studied, particularly for insect vectors of plant pathogens. We studied the combined effects of host acclimation and infection with a plant pathogenic symbiont on feeding behavior of Bactericera cockerelli, an oligophagous psyllid widespread in both crop and natural habitats that feeds primarily on Solanaceae and transmits an economically important plant pathogen, Candidatus Liberibacter solanacearum (CLso). We used a factorial design and the electrical penetration graphing technique to disentangle the effects of host acclimation, CLso infection, and psyllid haplotype on the within-plant feeding behavior of B. cockerelli during conspecific and heterospecific host switches. This approach allows to connect phenotypic plasticity with the role of B. cockerelli as a vector by quantifying the frequency and duration of behaviors involved in CLso transmission. We found significant reductions in multiple metrics of B. cockerelli feeding efficiency, exacerbated by infection with CLso, which could lead to reduced transmission of this pathogen. Psyllid genotype was also important; the Central haplotype exhibited less dramatic changes in feeding efficiency than the Western haplotype during heterospecific host switches. Our study shows that host acclimation and heterospecific host switching directly alter feeding behaviors underlying pathogen transmission, and that the magnitude of feeding efficiency reductions depends on both host genotype and infection status.

opencc-zeroMay 2024View details →

Adiponectin rescues synaptic plasticity in the dentate gyrus of a mouse model of fragile X syndrome

Fragile X Syndrome (FXS) is the most common inherited cause of intellectual disability and is the leading known single-gene cause of autism spectrum disorder. FXS patients display varied behavioural deficits that include mild to severe cognitive impairments in addition to mood disorders. Currently, there is no cure for this condition, however, there is an emerging focus on therapies that inhibit mTOR-dependent protein synthesis due to the clinical effectiveness of metformin for alleviating some behavioural symptoms in FXS. Adiponectin (APN) is a neurohormone that is released by adipocytes and provides an alternative means to inhibit mTOR activation in the brain. In these studies, we show that Fmr1 KO mice, like FXS patients, show reduced levels of circulating APN, and that both LTP and LTD in the DG (dentate gyrus) are impaired. Brief (20 min) incubation of hippocampal slices in APN (50 nM) was able to rescue both LTP and LTD in the DG and increased both the surface expression and phosphorylation of GluA1 receptors. These results provide evidence for reduced adiponectin levels in FXS playing a role in decreasing bidirectional synaptic plasticity and show that therapies that enhance adiponectin levels may have therapeutic potential for this and related conditions.

opencc-by-4.0Apr 2024View details →

Elasto-plastic residual stress analysis of selective laser sintered porous materials based on 3D-multilayer thermo-structural phase-field simulations

The supporting data and utilities from the publication "Elasto-plastic residual stress analysis of selective laser sintered porous materials based on 3D-multilayer thermo-structural phase-field simulations" are recorded in this dataset.  Non-isothermal phase-field simulations of SLS process on SS316L material and subsequent elasto-plastic calculations were performed to analyze the development of plastic deformation and residual stress in SLS produced components during the processing. The dependence of the fusion zone, residual stress and plastic strain on the processing parameters namely, Beam power (Unit: Watts) and Scan speed (Unit: mm/s) were investigated.  To promote FAIR research data principles, the processed simulation data from the thermo-elasto-plastic calculations for all the process parameter sets (hereby refered as P-v sets) are curated in this dataset. The raw temporal data obtained from the processing simulations and the elasto-plastic could not be included in this dataset due to its high volume. However, the corresponding raw data can be requested by contacting the creators of this dataset (Yangyiwei Yang: <a href="mailto:yangyiwei.yang@mfm.tu-darmstadt.de">yangyiwei.yang@mfm.tu-darmstadt.de</a> and Somnath Bharech: <a href="mailto:somnath.bharech@tu-darmstadt.de">somnath.bharech@tu-darmstadt.de</a>). This dataset includes:  <ul> <li><code>average_value.csv</code>: Contains average values of mechanical properties (such as residual stress, plastic strain) for the powder bed and the fused strut of all the process parameter sets.</li> <li><code>mesostructures_tep_sls.zip</code> : Contains resampled mesostructures obtained at the last time step of the SLS processing simulations with thermo-elasto-plastic calculations for the P-v sets reported in the aforementioned investigation. Nomenclature of the sub-directories indicating the P-v sets follows: <code>tep_<beam power>-<scan speed></code>. Each of these sub-directories contain the mesostructures from last time step of the thermo-elasto-plastic analysis of each of the four layer scans and is named as: <code>TP_layer{1..4}_output_final.e</code>. These files can be opened using Paraview v.5.8.1 or higher. The nodal values are explained as follows:</li> </ul> <table> <tbody> <tr> <td>Nodal value name</td> <td>Symbol</td> <td>Description</td> <td>Unit</td> </tr> <tr> <td>T</td> <td>\(T\)</td> <td>Temperature field normalized by \(T_M\)</td> <td>-</td> </tr> <tr> <td>c</td> <td>\(\rho\)</td> <td>Substance order parameter</td> <td>-</td> </tr> <tr> <td>eps_ij  </td> <td>\(\varepsilon\)</td> <td>Strain</td> <td>-</td> </tr> <tr> <td>epsp_ij</td> <td>\(\varepsilon^\text{pl}\)</td> <td>Plastic strain</td> <td>-</td> </tr> <tr> <td>peeq</td> <td>\(p_\text{e}\)</td> <td>Accumulated plastic strain</td> <td>-</td> </tr> <tr> <td>sigma_ij  </td> <td>\(\sigma\)</td> <td>Stress</td> <td>MPa</td> </tr> <tr> <td>vonmises</td> <td>\(\sigma_\text{e}\)</td> <td>von Mises stress  </td> <td>MPa</td> </tr> <tr> <td>u</td> <td>\(\mathbf{u}\)</td> <td>Displacement</td> <td>µm</td> </tr> </tbody> </table>

Supplemental Data for Marie Vogel's "Life is Plastic"-Poster, Roscoff 2024

Open the record for dataset details and reuse information.

Data from: Thermal plasticity in protective wing pigmentation is modulated by genotype and food availability in an insect model of seasonal polyphenism

Phenotypic variation in natural populations results from complex interactions between organisms and their changing environments. The environment shapes both phenotypic frequencies (during adaptation) and organismal phenotypes (through phenotypic plasticity). Developmental plasticity, in particular, refers to the phenomenon whereby an organism's phenotype depends on the environmental conditions during development. It can match phenotype to ecological conditions and help organisms to cope with environmental heterogeneity, including differences between alternating seasons. Experimental studies of developmental plasticity often focus on the impact of individual environmental cues and do not take explicit account of genetic variation. In contrast, natural environments are complex, comprising multiple variables with combined effects that are poorly understood and may vary among genotypes. We investigated the effects of multifactorial environments on the development of the seasonally plastic eyespots of Bicyclus anynana butterflies. Eyespot size depends on developmental temperature and is involved in alternative seasonal strategies for predator avoidance. In nature, both temperature and food availability undergo seasonal fluctuations. However, our understanding of how thermal plasticity in eyespot size varies in response to food availability and across genotypes remains limited. To address this, we investigated the combined effects of temperature (T; two levels: 20°C and 27°C) and food availability (N; two levels: control and limited) during development. We examined their impact on wing and eyespot size in adult males and females from multiple genotypes (G; 28 families). We found evidence of thermal and nutritional plasticity and temperature-by-nutrition interactions (significant TxN) on the size of eyespots in both sexes. Food limitation resulted in relatively smaller eyespots and tempered the effects of temperature. Additionally, we found differences among families for thermal plasticity (significant GxT effects), but not for nutritional plasticity (non-significant GxN effects) nor for the combined effects of temperature and food limitation (non-significant GxTxN effects). Our results reveal the context dependence of thermal plasticity, with the slope of thermal reaction norms varying across genotypes and across nutritional environments. We discuss these results in light of the ecological significance of pigmentation and the value of considering thermal plasticity in studies of the biological impact of climate change.

Data from: Dietary plasticity and broad North Atlantic origins inferred from bulk and amino acid-specific δ15N and δ13C favor killer whale range expansions into Arctic waters

Killer whales (Orcinus orca) occur seasonally in the eastern Canadian Arctic (ECA), where their range expansion associated with declining sea ice have raised questions about the impacts of increasing killer whale predation pressure on Arctic-endemic prey. We assessed diet and distribution of ECA killer whales using bulk and compound specific stable isotope analysis (CSIA) of amino acids (AA) of 54 skin biopsies collected from 2009-2020 around Baffin Island, Canada. Bulk ECA killer whale skin δ15N and δ13C values did not overlap with potential Arctic prey after adjustment for trophic discrimination, and instead reflected foraging history in the North Atlantic prior to their arrival in the ECA. Adjusted killer whale stable isotope (SI) values primarily overlapped with several species of North Atlantic baleen whales or tuna. Amino acid (AA)-specific δ15N values indicated the ECA killer whales fed primarily on marine mammals, having similar glutamic acid δ15N – phenylalanine δ15N (δ15NGlx-Phe) and threonine δ15N (δ15NThr) as mammal-eating killer whales from the eastern North Pacific (ENP) that served as a comparative framework. However, one ECA whale grouped with the fish-eating ENP ecotype based δ15NThr. Distinctive essential AA δ13C of ECA killer whale groups, along with bulk SI similarity to killer whales from different regions of the North Atlantic, indicate different populations converge in Arctic waters from a broad source area. Generalist diet and long-distance dispersal capacity favor range expansions, and integration of these insights will be critical for assessing ecological impacts of increasing killer whale predation pressure on Arctic-endemic species.

Data from: Phenotypically plastic responses to environmental variation are more complex than life history theory predicts

For insects that exhibit wing polyphenic development, abiotic and biotic signals dictate the adult wing morphology of the insect in an adaptive manner such that in stressful environments the formation of a flight-capable morph is favored and in low stress environments a flightless morph is favored. While there is a relatively large amount known about the environmental cues that dictate morph formation in wing polyphenic hemipterans like planthoppers and aphids, whether those cues dictate the same morphs in non-hemipteran (i.e. cricket) wing polyphenic species has not been explicitly investigated. To experimentally test the generality of environmental cue determination of wing polyphenism across taxa with diverse life histories, in this study we tested the importance of food quantity, parasitic infection, and tactile cues on wing morph determination in the wing polyphenic sand field cricket, Gryllus firmus. Our results also show that certain stress cues, such as severe diet quantity limitation and parasitic infection, actually led to an increase in the production of flightless morph. Based on these findings, our results suggest that physiological and genetic constraints are important to an organism's ability to respond to environmental variation in an adaptive manner beyond simple life history trade-offs.

Data from: Helper plasticity in response to breeder turnover in gray wolves

Nonbreeding helpers can greatly improve the survival of young and reproductive fitness of breeders in many cooperatively breeding species. Breeder turnover, in turn, can have profound effects on dispersal decisions made by helpers. Despite its importance in explaining group size and predicting population demography of cooperative breeders, our current understanding of how individual traits influence animal behavior after disruptions to social structure is incomplete particularly for terrestrial mammals. We used 12 years of genetic sampling and group pedigrees of gray wolves (Canis lupus) in Idaho, USA, to ask questions about how breeder turnover affected the apparent decisions by mature helpers (>2-year-old) to stay or leave a group over a one-year time interval. We found that helpers showed plasticity in their responses to breeder turnover. Most notably, helpers varied by sex and appeared to base dispersal decisions on the sex of the breeder that was lost as well. Male and female helpers stayed in a group slightly more often when there was breeder turnover of the same sex, although males that stayed were often recent adoptees in the group. Males, however, appeared to remain in a group less often when there was breeding female turnover likely because such vacancies were typically filled by related females from the males' natal group (i.e., inbreeding avoidance). We show that helpers exploit instability in the breeding pair to secure future breeding opportunities for themselves. The confluence of breeder turnover, helper sex, and dispersal and breeding strategies merge to influence group composition in gray wolves.