The seasonal reproduction of the three-spined stickleback is stimulated by long day photoperiod. As in other vertebrates, the reproductive system of stickleback is regulated by the brain-pituitary-gonadal (BPG) axis which is largely controlled by feedback effects. Both negative and positive feedback effects on the BPG axis have been found in fish. So far, the roles feedback effects on the BPG axis play in the photoperiodic regulation of seasonal reproduction are still unclear. This thesis focused on the photoperiodic regulation and gonadal feedback effects on the gene expressions of gonadotropin (GtH) and gonadotropin-releasing hormones (GnRH) in the brain and pituitary, and how gonadal feedback regulated the steroid homeostasis in stickleback.Both GnRH2 and GnRH3 mRNA was found in the hypothalamus. Higher expression levels of both GnRH2 and 3 in breeding than in post-breeding males suggested that they are both involved in seasonal reproduction. There was no evidence for a role of GnRH3, which may be the dominating form, in the photoperiodic control of reproduction. However, the polarity of the feedback effect on gnrh3 gene expression may turn from positive to be negative when the males went into post-breeding state. Tapeworm, Schistocephalus solidus, infection inhibited the reproduction of sticklebacks. However, the infection caused higher expression levels of both GnRHs and GtHs genes, which may be due to feedback effect on the BPG axis.Under short day, both lh-β and fsh-β were suppressed by low androgen levels. This negative feedback may inhibit maturation completely, unless a rise of androgens triggers positive feedback under long day. The change in feedback polarity may result in all or nothing maturation. Furthermore, the androgen inhibitory effect on lh-β and fsh-β under short day could be abolished by aromatase inhibitor, which means the estrogen may cause negative feedback in males under short day.There was no compensation effect on plasma androgen level in fully mature hemi-castrated fish. However, both testosterone and 11-ketoandrostenedione treatments increased plasma levels much less in sham-operated fish than in castrated ones, indicating that homeostatic mechanisms are nevertheless present. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows<strong>:  </strong>Paper 1: Submitted. Paper 3: Submitted. Paper 4: Submitted.</p>

Throughout their lives, prey organisms must balance the tradeoff between fitness-related activities and the risk of predation. To successfully mediate such tradeoffs, prey must have an accurate method to gauge current predation risk. For many aquatic organisms, the use of chemosensory information has been shown to be a ubiquitous and useful tool in mediating predation risk. The chemical cues to which aquatic organisms respond include the odour of known predators and the odour of a damaged conspecific or known or closely related heterospecific. In fishes, the response to damage-released cues from conspecifics or closely related heterospecifics has been shown to be innate, while the response to distantly related unknown heterospecific cues are likely learned. In a series of laboratory and field studies I examined the role of learning in the ability of fathead minnows to respond to damage-released cues of brook stickleback as an indication of predation risk. My results indicate that minnows from a population without stickleback do not recognize stickleback cues as dangerous. However, following the introduction of stickleback, minnows learn to recognize stickleback cues as dangerous. Further study indicated a low ratio of stickleback to minnows in a given population will decrease the likelihood of learning when compared with a similar sized population containing a higher ratio of stickleback to minnows. I also demonstrated that an increase in habitat complexity decreases the ability of minnows to learn to recognize stickleback cues. Studies have further demonstrated that in the face of predation (as indicated by chemical cues from minnows and stickleback) minnows will decrease their antipredator response when in the presence of a fish shoal, especially a shoal of conspecifics. Finally, an examination of the effects of a minnows length, body condition and breeding status indicate that morphological parameters can play a significant role in the intensity of response to heterospecific and conspecific damage-released cues. Previous works have demonstrated that behavioural responses to heterospecific damage-released cues increase the probability of the receiver surviving a predation event. This has important implications for predator/prey interactions and consequently factors affecting the learning and use of such cues deserve increasing attention from ecologists.

Throughout their lives, prey organisms must balance the tradeoff between fitness-related activities and the risk of predation. To successfully mediate such tradeoffs, prey must have an accurate method to gauge current predation risk. For many aquatic organisms, the use of chemosensory information has been shown to be a ubiquitous and useful tool in mediating predation risk. The chemical cues to which aquatic organisms respond include the odour of known predators and the odour of a damaged conspecific or known or closely related heterospecific. In fishes, the response to damage-released cues from conspecifics or closely related heterospecifics has been shown to be innate, while the response to distantly related unknown heterospecific cues are likely learned. In a series of laboratory and field studies I examined the role of learning in the ability of fathead minnows to respond to damage-released cues of brook stickleback as an indication of predation risk. My results indicate that minnows from a population without stickleback do not recognize stickleback cues as dangerous. However, following the introduction of stickleback, minnows learn to recognize stickleback cues as dangerous. Further study indicated a low ratio of stickleback to minnows in a given population will decrease the likelihood of learning when compared with a similar sized population containing a higher ratio of stickleback to minnows. I also demonstrated that an increase in habitat complexity decreases the ability of minnows to learn to recognize stickleback cues. Studies have further demonstrated that in the face of predation (as indicated by chemical cues from minnows and stickleback) minnows will decrease their antipredator response when in the presence of a fish shoal, especially a shoal of conspecifics. Finally, an examination of the effects of a minnows length, body condition and breeding status indicate that morphological parameters can play a significant role in the intensity of response to heterospecific and conspecific damage-released cues. Previous works have demonstrated that behavioural responses to heterospecific damage-released cues increase the probability of the receiver surviving a predation event. This has important implications for predator/prey interactions and consequently factors affecting the learning and use of such cues deserve increasing attention from ecologists.

Numerous contaminants are present in mixtures in the aquatic environment. Among these are the azoles, a group of chemicals that includes both pharmaceuticals and pesticides. Azole fungicides are designed to inhibit lanosterol 14-demethylase (cytochrome P450 (CYP) 51), while other azoles are intended to inhibit aromatase (CYP19), i.e. the enzyme catalyzing biosynthesis of estrogens. In fish, a variety of CYP enzymes are involved in biotransformation of waterborne contaminants, and in metabolism of endogenous compounds including steroidal hormones. The induction of CYP1A protein and 7-ethoxyresorufin O-deethylase (EROD) activity are common biomarkers for exposure to aryl hydrocarbon receptor (AhR) agonists in fish. We developed an assay to measure inhibition of CYP1A activity (EROD) in three-spined stickleback and rainbow trout gill tissue ex vivo. Several azole fungicides were found to be potent inhibitors of CYP1A activity. A wastewater effluent containing high concentrations of pharmaceuticals was also shown to inhibit CYP1A activity. Further, several azoles inhibited CYP19 activity in rainbow trout brain microsomes in vitro. Azole mixtures reduced both CYP1A and CYP19 activity monotonically and in an additive way. Given the additive action of the azoles, studies to determine adverse effects of azole mixtures on CYP-regulated physiological functions in fish are needed. Induction of EROD and of gene expression of CYP1 in several organs was observed in an in vivo exposure with the same effluent shown to inhibit EROD. This finding could imply that there was a mixture of AhR agonists and CYP1A inhibitors in the effluent. Finally, wastewater treatment technologies were evaluated using biomarker responses in rainbow trout exposed to effluents of different treatments. The results from chemical analysis together with the biomarker results show that ozone and granulated active carbon treatment removed most pharmaceuticals, as well as AhR agonists and other chemicals present in the regular effluent. This part of the thesis demonstrates that biomarkers in fish such as induction of CYP1 gene expression are applicable to evaluate the efficiency of different treatment technologies for wastewater.

Descent with modification (Darwin, 1859) overwhelmingly occurs through the process of natural selection on genetically variable traits. Following deglaciation in the Pleistocene, morphologically conservative marine stickleback (Gasterosteus aculeatus) from two distinct mitochondrial DNA lineages colonized freshwater habitats on the Haida Gwaii archipelago. These freshwater populations have radiated in response to a diverse selective landscape on the archipelago and exhibit morphological diversity that equals or exceeds the known range for the species in the circumboreal distribution. I investigated the body shape of 3808 stickleback from 125 isolated populations from Haida Gwaii using geometric morphometrics. Relative warp and discriminant function analysis were used to quantify lateral shape differences based on partial warp scores. which were generated from twelve homologous landmarks digitally placed on each specimen's image. The possibility of ontogenetic shifts and sexual dimorphism of body shape were examined for each population on the archipelago. Stickleback body-shape changes during ontogeny were highly variable, and cannot be predicted by the volume and water clarity of the habitat. I found a slight sexual dimorphism in adult body shape, the magnitude of which could not be predicted by habitat volume and clarity. A number of comparisons were made to elucidate the possible causes for selection for divergence of adult body shape among Haida Gwaii populations. Body shape of parapatric lake-river stickleback populations differed significantly in concordance with hydrodynamic principles. Comparing the body shape of Haida Gwaii populations to the shape of sympatric benthic-limnetic species pairs from southwestern British Columbia. I found that benthic and limnetic body-shape ecotypes differ by up to 56% of the total variation among Haida Gwaii populations. Tests for phenotypic plasticity of body shape were conducted with two morphologically distinct stickleback populations that had been transplanted into two separate experimental ponds that were the ecological opposites of the respective source lakes. I found evidence for some phenotypic plasticity in body shape; the difference between each source and experimental population was approximately 11% of the total variation in body shape among populations throughout the archipelago. Throughout the islands adult body shape and size can be predicted by both abiotic and biotic factors of the habitat. Populations with derived shape (CV1+), including thicker peduncles, posteriad and closely spaced dorsal spines, anteriad pelvis, short dorsal and anal fins, and smaller body size occur in small, shallow, stained ponds, and populations with less derived shape (CVI-). with smaller narrow peduncles. anteriad and widely spaced dorsal spines. posteriad pelvis. longer dorsal and anal fins, and larger size occur in large, deep. clear lakes. There were large-bodied populations with derived shape (CV2-). including smaller heads and shallower elongate bodies in open water habitats of low productivity, and populations with smaller size and less derived shape (CV2±), with larger heads and deeper bodies, in higher productivity, structurally complex habitats. Populations with robust defensive adaptations have less derived shape (CV1-) and larger size, in response to salmonid predation, while populations with weak defences had derived shape (CV 1+), in response to bird/invertebrate predators. The ecomorphological relationships were consistent between mitochondrial lineages and replicated in each geographical region on the archipelago among geographically distant populations, suggesting the parallel evolution of body shape governed by the hydrodynamic constraints of each habitat. Although initially colonized by ancestors with conserved morphology, the stickleback populations on Haida Gwaii exhibit body shapes suited to the hydrodynamic landscape of the habitat, demonstrating the predictability of natural selection in adaptive radiations.

No description available.

Dietary restriction (DR), is a reduction in food intake, either through overall calorie or specific macronutrient intake, while avoiding malnutrition. DR has been consistently shown to increase longevity and protect against age related diseases. Although originally thought to be the result of a reduction in caloric intake, recent evidence suggests that the ratio of macronutrients, particularly that of protein : non-protein energy, also plays a role. The broad range of species in which DR is known to be effective, suggests an evolutionary conserved mechanism. However, the suggestion of a strong model species bias and a potential sex bias have led some to question the ubiquity of responses to DR. Here, I address the following questions: (i) How consistent is the effect of DR on reproduction? (ii) How does varying macronutrient intake effect both growth and body composition in three-spine sticklebacks (Gasterosteus aculeatus)? (iii) What is the effect of dietary macronutrient intake on lifespan and reproduction in the three-spine stickleback? and (iv) How does changing macronutrient intake impact fitness related traits, such as condition and performance in three-spine sticklebacks? Through use of a systematic review and meta-analysis, I show that the effect of DR on reproduction is evolutionarily conserved, though the effect is stronger in model species. However, when accounting for all significant moderators there is no evidence of the suggested sex differences in the effect of DR. I show that body composition is predicted by dietary lipid intake, with sticklebacks targeting a lower ratio of protein : fat within the body, potentially via metabolism and excretion of protein. These results hint at a link between conversion and excretion of protein and survival costs associated with high protein diets. I show that mortality risk is reduced at balanced protein : lipid intakes in males and generally at low protein : lipid intakes for females. However, the effect in females is not consistent throughout life. I further show that reproduction is maximised on high protein : lipid intakes for both sexes. These results suggest a macronutrient mediated trade-off between lifespan and reproduction in male three-spine sticklebacks. Finally I show a positive effect of lipid intake on male condition (a possible indicator of overall health) hinting at a relationship between lipid intake, adiposity, health and lifespan in male three-spine sticklebacks.

xvi, 112 p. : ill. (some col.) / In seasonal environments, the ability to take advantage of the favorable seasons and avoid or mitigate the effects of the unfavorable ones is essential for organismal fitness. Many polar and temperate organisms use photoperiod (length of day) to time seasonal life history events because photoperiod's regular annual cycle makes it a very reliable indicator of seasonality. This reliability allows organisms to anticipate and properly prepare for seasonal change. Although photoperiodism is widespread in polar and temperate vertebrates, little is known relative to invertebrates regarding how its use varies with environment and this method's underlying genetic and physiological basis. This dissertation is focused on demonstrating the proper methodology for the study of photoperiodism and establishing the threespine stickleback as a model of vertebrate photoperiodism. Chapter I is an introduction to photoperiodism, how it is influenced by environment, the physiological basis of its output, and a summary of the chapters that follow. Chapter II explains an analytical method to test for causality and applies this method to data that have been interpreted as evidence that the circadian clock is causally involved in photoperiodism. Chapter III describes the photoperiodic response of threespine stickleback Gasterosteus aculeatus populations from two latitudes. These results are used to inform an empirical examination of the previously described assertion that the circadian clock is causally involved in photoperiodism. Chapter IV examines the physiological basis of early photoperiodic response using the threespine stickleback as a model teleost fish. Chapter V summarizes the previous chapters, describes their significance, and suggests future research directions. This dissertation includes both previously published and co-authored material. Supplementary Excel files demonstrating the analyses used in Chapter III are also included in this dissertation. / Committee in charge: Eric Johnson, Chairperson; William Cresko, Advisor; William Bradshaw, Member; Judith Eisen, Member; Patricia McDowell, Outside Member

Across vertebrates, skeletal shapes are diverse, and much of this variation appears to be adaptive. In contrast, the early developmental programs of these structures are highly conserved across vertebrates. The question then becomes where in the conserved genetic programs of skeletal development does variation lie to direct diversity? In threespine stickleback, rapid changes in head and body shape have been documented during repeated and independent invasions of oceanic fish into freshwater habitats in regions deglaciated approximately 13,000 years ago. However, recent research indicates that similar phenotypic and genetic divergence can occur in decades. A remaining challenge is to link stickleback population genomic variation to causal genes that underlie such rapid phenotypic evolution. Here I use genome wide association studies (GWAS) in natural populations of stickleback to uncover genomic regions that contribute to variation of two dermal bone derived traits, lateral plate number and opercle shape. The decrease of lateral plate body armor and change in opercle bone shape, important for feeding mechanics, are classically associated with freshwater divergence. GWAS has recently begun to be used in natural populations but is still under scrutiny for performance among different populations. Using a population of phenotypically variable stickleback in Oregon, GWAS proved an effective method to uncover genomic regions and genetic variants known to contribute to lateral plate number and opercle shape, as well as new genomic regions and candidate genes not previously implicated in phenotypic variation. Although successful, using similar methods on decades old stickleback populations in Alaska revealed the challenges that accompany controlling population structure created by strong natural selection. Together, I found that although lateral plate number and opercle shape rapidly evolve in a coordinated fashion during adaptation from marine to freshwater environments, phenotypic variation is largely driven by independent genetic architectures. However, in very rapidly evolving populations, despite this independence of genetic architecture, the genetic variants contributing to the traits co-localize to similar genomic regions. This finding could be either biological or methodological which highlights the promise and limitations of using GWAS to identify genetic variation that gives rise to phenotypic diversity. This dissertation includes unpublished co-authored material.

Evolution shapes ecosystems but the processes by which this occurs are not well understood. Adaptive change in resource expensive traits may underlie one such process, as evolution altering a species’ resource needs may effect how that species interacts with ecosystem resources. For this, Ecological Stoichiometry (ES) may be a tractable framework, as it simplifies organisms into elemental ratios and then applies mass-balance to predict changes in diet and waste interactions. ES detects variation in resource expensive traits as variation in elemental ratios, and predicts compensation via parallel changes in diet (e.g. high phosphorous individuals consume high phosphorus diets) and/or offsetting changes in waste (e.g. high phosphorous individuals release low phosphorus waste). To test the utility of this framework and improve our understanding of eco-evolutionary dynamics, I studied variation in phenotypic traits, genetics, elemental content and resource interactions within and across natural populations of highly regarded eco-evolutionary model species threespine stickleback (Gasterosteus aculeatus). First, I related heritable variation in phosphorus rich bony traits and genetics commonly under natural selection with variation in elemental content (N:P) to determine the magnitude and basis of intraspecific variation in N:P. Second, I investigated the ecosystem consequences of variation in elemental content by determining whether stickleback compensate through changes in diet choice and excretion rates. I found stickleback vary widely in elemental composition (3.0 – 9.4:1 N:P) which models explained well with four bone related traits: bone mineralization, body size, lateral plating and pelvis size (R2 > 0.52). Additional genetic models linked variation in Eda alleles (which underlie lateral plating) with a 12% shift in stickleback N:P. Stickleback compensated for this variation in N:P demand by altering diet choice rather than excretion rates, and by maximizing dietary inputs through changes in gut morphology. Within and across populations, high phosphorus stickleback consumed a larger proportion of high phosphorus prey and contained longer gastrointestinal tracts that more efficiency process diet resources. These results demonstrate that heritable variation in elemental composition is ecologically relevant with individual traits and genetics having large effects. As individuals compensated by altering resource acquisition rather than release, the direct ecological consequences of evolutionary change in these resource expensive traits is likely larger for food web structure and abundance than nutrient dynamics. / Graduate / 2018-12-19