Adult Sticklebacks Research Articles

Assessing the potential for successful translocation and co‐management of two endangered aquatic species

AbstractTranslocation programmes for endangered species typically focus on a single species, but in areas where little native habitat remains, it may be necessary to translocate multiple species to the same sites. Interactions between translocated species, such as predation and competition, are among the factors that need to be considered when planning multispecies translocations. Translocation sites for aquatic species are particularly scarce in southern California, where a limited number of sites exist for historically co‐occurring endangered mountain yellow‐legged frogs Rana muscosa and unarmoured three‐spine sticklebacks Gasterosteus aculeatus williamsoni. To determine how these species would interact if translocated to the same sites, we carried out experiments ex situ with R. muscosa tadpoles and a surrogate subspecies of stickleback (G. a. microcephalus). We found that (1) adult sticklebacks preyed on hatchling tadpoles but did not consume R. muscosa eggs or large tadpoles; (2) tadpoles did not consume stickleback eggs or disturb sticklebacks nests; and (3) both species' microhabitat use shifted slightly when the other was present. Our results suggest that these species can likely be co‐managed successfully, if measures are taken to curb stickleback predation on tadpoles until the R. muscosa population is well established. Using ex situ studies to evaluate species interactions prior to translocation is an approach that could prove useful in other species recovery programmes. Multispecies translocations could make better use of available resources when habitat is limited and promote ecosystem recovery by re‐establishing interactions among native species.

Predator life history and prey ontogeny limit natural selection on the major armour gene,Eda, in threespine stickleback

AbstractNatural selection shapes the evolution of antipredator traits in prey. However, selection in the wild depends on ecological context, including features of predator and prey populations, making field studies of selection critical to understanding how predators shape selection on prey defences. Threespine stickleback (Gasterosteus aculeatus) is a classic system to study the effects of predators on the natural selection of prey. In lakes and rivers, fish predators have been shown to impose selection against low plated adult stickleback phenotypes and genotypes. We directly measured selection by predatory salmonids on theEctodysplasin‐A(Eda) gene in estuary stickleback from California. Despite previous studies showing a positive correlation between predator presence and frequency of theEda“complete” allele in estuary populations, we found thatEda“low” genotypes were not significantly more frequent in salmonid predator diets. Further, we found no evidence of changes inEdagenotype frequencies across generations that would suggest directional selection driven by predators. Prior selection studies have examined the effects of large resident trout on adult stickleback. In contrast, predators in this study were juvenile anadromous salmonids, which only ate juvenile stickleback whose plate phenotypes had not fully developed. Thus, in this case, predator life history and stickleback ontogeny may preclude strong selection on stickleback armour. Our results underscore the importance of selection studies in the wild for understanding the context‐dependent nature of selection in natural populations.

Differences in the contributions of sex linkage and androgen regulation to sex-biased gene expression in juvenile and adult sticklebacks.

Different evolutionary interests between males and females can lead to the evolution of sexual dimorphism. However, intersex genetic correlations due to the shared genome can constrain the evolution of sexual dimorphism, resulting in intra-locus sexual conflict. One of the mechanisms resolving this conflict is sex linkage, which allows males and females to carry different alleles on sex chromosomes. Another is a regulatory mutation causing sex-biased gene expression, which is often mediated by gonadal steroids in vertebrates. How do these two mechanisms differ in the contributions to the resolution of intra-locus sexual conflict? The magnitude of sexual conflict often varies between the juvenile and adult stages. Because gonadal steroids change in titre during development, we hypothesized that gonadal steroids play a role in sexual dimorphism expression only at certain developmental stages, whereas sex linkage is more important for sexual dimorphism expressed throughout life. Our brain transcriptome analysis of juvenile and adult threespine sticklebacks showed that the majority of genes that were sex-biased in both stages were sex-linked. The relative contribution of androgen-dependent regulation to the sex-biased transcriptome increased and that of sex linkage declined in adults compared to juveniles. The magnitude of the sex differences was greater in sex-linked genes than androgen-responsive genes, suggesting that sex linkage is more effective than androgen regulation in the production of large sex differences in gene expression. Overall, our data are consistent with the hypothesis that sex linkage is effective in resolving sexual conflict throughout life, whereas androgen-dependent regulation can contribute to temporary resolution of sexual conflict.

Sodium perchlorate induces non-alcoholic fatty liver disease in developing stickleback

Perchlorate is a pervasive, water-soluble contaminant that competitively inhibits the sodium/iodide symporter, reducing the available iodide for thyroid hormone synthesis. Insufficient iodide uptake can lead to hypothyroidism and metabolic syndromes. Because metabolism, obesity and non-alcoholic fatty liver disease (NAFLD) are tightly linked, we hypothesized that perchlorate would act as an obesogen and cause NAFLD via accumulation of lipids in liver of developing threespine stickleback (Gasterosteus aculeatus). We performed an upshift/downshift exposure regime (clean water to perchlorate treated water or perchlorate treated water to clean water) on stickleback embryos at two concentrations (30 mg/L and 100 mg/L) plus the control (0 mg/L) over the course of 305 days. Adult stickleback were euthanized, H&E stained and analyzed for liver morphology. Specifically, we counted the number of lipid droplets, and measured the area of each droplet and the total lipid area of a representative section of liver. We found that perchlorate treated fish had more and larger lipid droplets, and a larger percentage of lipid in their liver than control fish. These data indicate that perchlorate causes NAFLD and hepatic steatosis in stickleback at concentrations commonly found at contaminated sites. These data also indicate the potential of perchlorate to act as an obesogen. Future studies should investigate the obesogenic capacity of perchlorate by examining organ specific lipid accumulation and whether perchlorate induces these effects at concentrations commonly found in drinking water. Work is also needed to determine the mechanisms by which perchlorate induces lipid accumulation.

The effect of top-predator presence and phenotype on aquatic microbial communities.

The presence of predators can impact a variety of organisms within the ecosystem, including microorganisms. Because the effects of fish predators and their phenotypic differences on microbial communities have not received much attention, we tested how the presence/absence, genotype, and plasticity of the predatory three‐spine stickleback (Gasterosteus aculeatus) influence aquatic microbes in outdoor mesocosms. We reared lake and stream stickleback genotypes on contrasting food resources to adulthood, and then added them to aquatic mesocosm ecosystems to assess their impact on the planktonic bacterial community. We also investigated whether the effects of fish persisted following the removal of adults, and the subsequent addition of a homogenous juvenile fish population. The presence of adult stickleback increased the number of bacterial OTUs and altered the size structure of the microbial community, whereas their phenotype affected bacterial community composition. Some of these effects were detectable after adult fish were removed from the mesocosms, and after juvenile fish were placed in the tanks, most of these effects disappeared. Our results suggest that fish can have strong short‐term effects on microbial communities that are partially mediated by phenotypic variation of fish.

Perchlorate Exposure Reduces Primordial Germ Cell Number in Female Threespine Stickleback.

Perchlorate is a common aquatic contaminant that has long been known to affect thyroid function in vertebrates, including humans. More recently perchlorate has been shown to affect primordial sexual differentiation in the aquatic model fishes zebrafish and threespine stickleback, but the mechanism has been unclear. Stickleback exposed to perchlorate from fertilization have increased androgen levels in the embryo and disrupted reproductive morphologies as adults, suggesting that perchlorate could disrupt the earliest stages of primordial sexual differentiation when primordial germ cells (PGCs) begin to form the gonad. Female stickleback have three to four times the number of PGCs as males during the first weeks of development. We hypothesized that perchlorate exposure affects primordial sexual differentiation by reducing the number of germ cells in the gonad during an important window of stickleback sex determination at 14–18 days post fertilization (dpf). We tested this hypothesis by quantifying the number of PGCs at 16 dpf in control and 100 mg/L perchlorate-treated male and female stickleback. Perchlorate exposure from the time of fertilization resulted in significantly reduced PGC number only in genotypic females, suggesting that the masculinizing effects of perchlorate observed in adult stickleback may result from early changes to the number of PGCs at a time critical for sex determination. To our knowledge, this is the first evidence of a connection between an endocrine disruptor and reduction in PGC number prior to the first meiosis during sex determination. These findings suggest that a mode of action of perchlorate on adult reproductive phenotypes in vertebrates, including humans, such as altered fecundity and sex reversal or intersex gonads, may stem from early changes to germ cell development.

Small Variations in Early-Life Environment Can Affect Coping Behaviour in Response to Foraging Challenge in the Three-Spined Stickleback.

ContextAn increasing concern in the face of human expansion throughout natural habitats is whether animal populations can respond adaptively when confronted with challenges like environmental change and novelty. Behavioural flexibility is an important factor in estimating the adaptive potential of both individuals and populations, and predicting the degree to which they can cope with change.Study DesignThis study on the three-spined stickleback (Gasterosteus aculeatus) is an empiric illustration of the degree of behavioural variation that can emerge between semi-natural systems within only a single generation. Wild-caught adult sticklebacks (P, N = 400) were randomly distributed in equal densities over 20 standardized semi-natural environments (ponds), and one year later offspring (F1, N = 652) were presented with repeated behavioural assays. Individuals were challenged to reach a food source through a novel transparent obstacle, during which exploration, activity, foraging, sociability and wall-biting behaviours were recorded through video observation. We found that coping responses of individuals from the first generation to this unfamiliar foraging challenge were related to even relatively small, naturally diversified variation in developmental environment. All measured behaviours were correlated with each other. Especially exploration, sociability and wall-biting were found to differ significantly between ponds. These differences could not be explained by stickleback density or the turbidity of the water.FindingsOur findings show that a) differences in early-life environment appear to affect stickleback feeding behaviour later in life; b) this is the case even when the environmental differences are only small, within natural parameters and diversified gradually; and c) effects are present despite semi-natural conditions that fluctuate during the year. Therefore, in behaviourally plastic animals like the stickleback, the adaptive response to human-induced habitat disturbance may occur rapidly (within one generation) and vary strongly based on the system’s (starting) conditions. This has important implications for the variability in animal behaviour, which may be much larger than expected from studying laboratory systems, as well as for the validity of predictions of population responses to change.

Experimental Evidence of an Eco-evolutionary Feedback during Adaptive Divergence

Differences in how organisms modify their environment can evolve rapidly and might influence adaptive population divergence. In a common garden experiment in aquatic mesocosms, we found that adult stickleback from a recently diverged pair of lake and stream populations had contrasting effects on ecosystem metrics. These modifications were caused by both genetic and plastic differences between populations and were sometimes comparable in magnitude to those caused by the presence/absence of stickleback. Lake and stream fish differentially affected the biomass of zooplankton and phytoplankton, the concentration of phosphorus, and the abundance of several prey (e.g., copepods) and non-prey (e.g., cyanobacteria) species. The adult-mediated effects on mesocosm ecosystems influenced the survival and growth of a subsequent generation of juvenile stickleback reared in the same mesocosms. The prior presence of adults decreased the overall growth rate of juveniles, and the prior presence of stream adults lowered overall juvenile survival. Among the survivors, lake juveniles grew faster than co-occurring stream juveniles, except in mesocosm ecosystems previously modified by adult lake fish that were reared on plankton. Overall, our results provide evidence for reciprocal interactions between ecosystem dynamics and evolutionary change (i.e., eco-evolutionary feedbacks) in the early stages of adaptive population divergence.

First records of three-spined stickleback Gasterosteus aculeatus in Svalbard freshwaters: An effect of climate change?

Several fish species, including three-spined stickleback, Gasterosteus aculeatus, have shown a more northerly distribution in the Norwegian Sea into the eastern part of the Arctic Ocean during the last decades. Three-spined stickleback has however never been recorded in freshwaters on Svalbard, whereas Arctic charr, Salvelinus alpinus, has been the only reproducing fish with anadromous, resident, and landlocked populations. On this background, we sampled fish by gillnetting, electrofishing, and trapping in two watercourses on the island of Spitsbergen, Svalbard, during the summers 2001 and 2006. During these surveys, three-spined sticklebacks were captured in both localities. To our knowledge, this represents the first records of the species in Svalbard freshwaters. The presence of a few adult sticklebacks does not provide evidence that breeding populations are established. However, increased temperatures associated with climate change in recent years in both Arctic marine and freshwater environments may have facilitated the appearance of the species in these localities and thus may be a first indication of the potential establishment of this species in Svalbard freshwaters.

Perchlorate exposure does not modulate temporal variation of whole-body thyroid and androgen hormone content in threespine stickleback

Previously we showed that exposure of threespine stickleback (Gasterosteus aculeatus) to the endocrine disruptor perchlorate results in pronounced structural changes in thyroid and gonad, while surprisingly, whole-body thyroid hormone concentrations remain unaffected. To test for hormone titer variations on a finer scale, we evaluated the interactive effects of time (diel and reproductive season) and perchlorate exposure on whole-body contents of triiodothyronine (T3), thyroxine (T4), and 11-ketotestosterone (11-KT) in captive stickleback. Adult stickleback were exposed to 100ppm perchlorate or control water and sampled at 4-h intervals across the 24-hday and at one time-point (1100h) weekly across the reproductive season (May–July). Neither whole-body T3 nor T4 concentration significantly differed across the day in control or perchlorate treated stickleback. Across the reproductive season, whole-body T3 concentration remained stable while T4 significantly increased. However, neither hormone concentration was significantly affected by perchlorate, verifying our previous studies. The concentration of whole-body 11-KT, a major fish androgen, displayed significant diel variation and also steadily declined across the reproductive season in untreated males; perchlorate exposure did not influence the concentration of 11-KT in either diel or reproductive season schedules. Diel and reproductive season variations in 11-KT content in male stickleback are likely related to reproductive physiology and behavior. The observed increase in T4 content across the reproductive season may be reflective of increased energy investment in reproduction near the end of the life cycle.

Behavioural Disturbances in a Temperate Fish Exposed to Sustained High-CO2 Levels

As atmospheric CO2 levels rise, the CO2 concentration in ocean surface waters increases through a process commonly referred to as ocean acidification. Recently, surprising behavioural modifications has been detected in the early life stages of tropical coral reef fish exposed to ocean acidification-relevant CO2 concentrations, but it has been unclear if this effect could occur in temperate waters. Here we show several severe behavioural disturbances, including effects on boldness, exploratory behaviour, lateralisation, and learning in a temperate fish, the three-spined stickleback (Gasterosteus aculeatus). The behavioural effects were consistent throughout the exposure period and increased in effect size with exposure time. We observed the effects on adult sticklebacks, a species known to be tolerant to other environmental stressors. Our findings suggest that behavioural abnormalities that stem from CO2 exposure are not restricted to sensitive tropical species or early life stages and may therefore affect fish on a global scale. The severity of disturbances and the possibility of a serious behavioural problem for fish across the globe is cause for concern.

Prolonged life span among endemicGasterosteuspopulations

Throughout their circumboreal coastal distribution, the threespine stickleback ( Gasterosteus aculeatus L., 1758) typically reproduce at 1 or 2 years of age (second or third summer) and die during the year of the reproductive cycle. Extending from a previous study on the Haida Gwaii archipelago, western Canada, that identified an exceptionally long life span (8 years) in a population of large-bodied threespine stickleback, we use pelvic spine annuli to examine age of the five largest adult stickleback of 12 additional populations comprising five populations with average-sized adults (45–60 mm standard length (SL)) and seven populations with large-bodied adult stickleback (>75 mm SL). Each of the small-bodied populations had a maximum age of 1 or 2 years typical for the taxon. Among the large-bodied populations, which also reached adult size in the 2nd year, adult stickleback in the populations ranged from 3 to 6 years, indicating extended longevity. Low productivity habitats and refuge against gape-limited piscivores, each of which theoretically predicts reduced rate of senescence, are associated with the greatest longevity among these populations. These data combined with the recent full genome sequence for stickleback provide opportunities for locating genetic markers for extended longevity.

Effect of prochloraz fungicide on biotransformation enzymes and oxidative stress parameters in three-spined stickleback (Gasterosteus aculeatus L.)

The aim of this study was to characterize biomarker responses in three-spined sticklebacks exposed to prochloraz (Pcz). For this purpose, adult sticklebacks were exposed for 2 weeks to prochloraz at 0, 10, 50, 100 and 500 μg/L prior to one week of depuration in clean water. At days 7, 14 and 21, several hepatic biomarkers were measured including 7-ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), glutathione peroxidase (GPx), catalase (CAT), total glutathione (GSH) content and thiobarbituric acid reactive substances (TBARS). Pcz induced a transient increase of antioxidant enzymes and a depletion of glutathione content during the first 7 days of exposure. This study showed that EROD activity and antioxidants were disrupted in a transient manner. GST was rapidly induced in a dose-dependent manner and this induction was persistent and observed also after depuration. GST appeared as a valuable biomarker to assess the exposure to Pcz.

The optimal foraging strategy of its stickleback host constrains a parasite's complex life cycle

AbstractThe cestode parasite Schistocephalus solidus' growth is limited by the size of its second intermediate host, the three-spined stickleback, Gasterosteus aculeatus. S. solidus should thus prefer a large stickleback as host. Since the stickleback is a predator of the parasite's previous intermediate host, a small copepod, the stickleback that consumes the infected copepod will probably be of a size for which this copepod has the optimal prey size. The optimal foraging decision of the stickleback may or may not be compatible with the parasite's preference. Infected copepods are present in early summer when both many size classes of young of the year and adult sticklebacks are potential predators. We offered laboratory bred three-spined sticklebacks of four size classes individually the choice among five prey types: two size classes of copepods, two classes of Daphnia of corresponding size as alternative prey and a third Daphnia size class that was larger than the larger copepod. We found that small copepods, the potential hosts of S. solidus, were most accepted by the smallest sticklebacks of about 1.5 cm of length, larger fish consumed a decreasing proportion; fish larger than 3.8 cm did not consume them at all. Experience with copepods over several weeks increased the acceptance for this prey to some extend but hardly in the largest fish. This suggests that S. solidus will end up usually in sticklebacks that are too small for the parasite so that it has to allow its host's further growth after infection to reach its definitive size.

Copper-induced oxidative stress in three-spined stickleback: relationship with hepatic metal levels

The aim of this study was to characterise biomarker responses in three-spined sticklebacks exposed to copper. For this purpose, adult sticklebacks were exposed for 3 weeks to copper sulphate at 0, 25, 100 and 200 μg L −1 as Cu. At days 4, 8, 12 and 21, several parameters were measured including liver, gonad and spleen somatic indexes, hepatic biomarkers (catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione (GSH), glutathione- S-transferase (GST) and 7-ethoxyresorufin-O-deethylase (EROD)) and hepatic copper and zinc concentrations. Copper induced a rapid and transient increase of antioxidant enzymes and a depletion of glutathione content during the first 8 days of exposure. Significant copper and zinc accumulation in fish liver were observed for the two higher exposure concentrations after 8 and 12 days, respectively. This study showed that copper induced an oxidative stress in fish liver before significant metal accumulation in the liver could be detected, suggesting the involvement of differential mechanisms in copper uptake and metabolism. Three-spined stickleback appears to be a sensitive model to study oxidative stress induced by metals.

Dietary differences between phenotypes with symmetrical and asymmetrical pelvis in the stickleback Gasterosteus aculeatus

Differential parasitism among phenotypes within populations can result from intrinsic factors such as immunocompetence or extrinsic factors such as ecological overlap with pathogens. In a recent study of a population of sticklebacks (Gasterosteus aculeatus) from Boulton Lake, British Columbia, relative cestode infections were related to pelvic-girdle symmetry. Adult sticklebacks with an asymmetrical pelvis had a higher incidence of infections than fish with a symmetrical pelvis, yet the pattern was reversed among yearlings. In the current study we test whether this unexpected result for yearlings, which is inconsistent with general theory coupling asymmetry with reduced immuno competence, might be due to ecological factors rather than to differences in immunocompetence. We analyze the diet of 9089 uninfected sticklebacks collected during a 15-year study and show that male and female yearling sticklebacks with an asymmetrical pelvis are more benthic than symmetrical phenotypes. This could reduce the probability of exposure to pelagic copepods, the primary hosts of the cestode Schistocephalus solidus, and thereby lead to lower infection rates relative to pelagic fish. This finding would account for the unusual reversal in relative parasitism in this population and raises the possibility of finding further diet-induced causes of differential parasitism in studies of asymmetry.

Characterization and localization of gonadotropin-releasing hormone in the brain and pituitary of the three-spined stickleback, Gasterosteus aculeatus

Radioimmunoassay (RIA) studies on highperformance liquid chromatography (HPLC) fractions of brain extracts of the three-spined stickleback, Gasterosteus aculeatus, provided evidence for at least two forms of gonadotropin-releasing hormone (GnRH). One form showed chromatographic and immunological properties similar to that of synthetic salmon GnRH (sGnRH). A second, unidentified form of GnRH eluted in the same position as chicken GnRH I (cGnRH-I); however, it did not cross-react in a cGnRH-I RIA. Furthermore, it cannot be excluded that chicken GnRH II (cGnRH-II) and maybe one other unidentified form are present in the stickleback. The distribution of GnRH in the brain of breeding adult male sticklebacks was studied by use of immunohistochemistry. Two antisera against sGnRH and antisera against mGnRH and cGnRH-II were applied on cryosections and visualized using the peroxidase-antiperoxidase method. Staining patterns were similar after incubations with all four antisera. Immunoreactive fibers were found in most parts of the brain. Three distinct groups of GnRH-immunoreactive perikarya were found in the nucleus olfactoretinalis, in the nucleus anterior periventricularis, and in the nucleus lateralis tuberis. Moreover, weakly stained cells occurred in a periventricular position in the midbrain. The proximal pars distalis of the pituitary, housing the gonadotropic cells, was richly innervated by GnRH-positive fibers. In the pars intermedia and in the rostral pars distalis, immunoreactive fibers were absent.

UNDERSTANDING THE EVOLUTION OF BEHAVIOR IN THREESPINE STICKLEBACK: THE VALUE OF GEOGRAPHIC VARIATION

AbstractThe endemic radiation of the threespine stickleback in North America possesses features that will permit us to gain unique insights into evolutionary pattern and process. Already, comparison of populations has demonstrated a remarkable level of adaptive differentiation in reproductive behavior across populations in both southern British Columbia, Canada, and the Cook Inlet Region of Alaska, U.S.A. In small, shallow lakes, threespine stickleback feed predominantly on benthic invertebrates in the shallow littoral zone. They form large groups that attack nests defended by males, and consume all of the young within. In contrast, in larger, more oligotrophic lakes, adult stickleback feed primarily on plankton in large open-water foraging groups. They do not form groups that cannibalize young in nests. In these non-cannibalistic populations, males perform more conspicuous courtship behavior than do those in cannibalistic populations, apparently because risk of attack by visually hunting conspecifics is low. Courtship in these lakes more often includes the conspicuous zig-zag dance, and less often, meandering and dorsal pricking. Anadromous populations tend to be cannibalistic, but also are often behaviorally intermediate. They incorporate the zig-zag dance in courtship more often than do males in cannibalistic freshwater populations, but also usually incorporate the less conspicuous dorsal pricking behavior. These results suggest that the ancestral condition was intermediate and that both cannibalistic and non-cannibalistic freshwater populations display some derived elements in their reproductive repertoires. This kind of comparison among populations thought to have been independently derived from the marine form can provide insights into the patterns and causes of adaptive diversification. For populations such as these, carefully chosen from disparate geographic locations so as to preclude common evolutionary history after colonization of freshwater,

Developmental changes in the brain-stem serotonergic nuclei of teleost fish and neural plasticity.

1. During early ontogeny, the serotonergic neurons in the brain stem of the three-spined stickleback shows a temporal and spatial developmental pattern that closely resembles that of amniotes. 2. However, in the adult fish, only the midline nuclei of the rostral group (dorsal and median raphe nuclei) and the dorsal lateral tegmental nucleus are consistently serotonin-immunoreactive (5-HTir), whereas the groups of the upper and lower rhombencephalon (raphe pontis, raphe magnus, and raphe pallidus/obscurus nuclei) are variable and, when present, contain relatively small numbers of 5-HTir neurons. 3. Using specific antisera against tryptophan 5-hydroxylase and aromatic L-amino acid decarboxylase, we have shown that the lateral B9 group and the groups of the upper and lower rhombencephalon are consistently present in adult sticklebacks. The results are discussed in relation to other known instances of neurotransmitter plasticity or transient neurotransmitter expression in teleost fish. 4. While there are several instances of transient expression of neurotransmitter markers by discrete neuronal populations, there is so far no evidence of changes from one neurotransmitter phenotype to another in the brain of teleost fish. However, there are indications of plasticity of expression of catecholamines and indoleamines, and their respective synthesizing enzymes, as reflected in age-dependent changes and variation between individuals of different physiological status. 5. As the brain grows continuously in teleost fish, and new neurons are added from proliferative regions, synaptic connections may be expected to undergo remodeling in all brain regions throughout life. Thus, the teleostean brain may be considered a suitable model for experimental studies of different aspects of neural plasticity.

Development of tyrosine hydroxylase-, dopamine- and dopamine β-hydroxylase-immunoreactive neurons in a teleost, the three-spined stickleback

The development of catecholaminergic neuronal systems in the brain of a teleost, the three-spined stickleback, was studied through embryonic to early larval stages by immunocytochemistry using specific antibodies against dopamine, tyrosine hydroxylase and dopamine β-hydroxylase. By analysing the spatiotemporal patterns of development for the catecholaminergic nuclei, possible homologies with nuclei in amniote brains have been identified. The noradrenergic neurons in the isthmus region of the rostral rhombencephalon originate in the same manner as the A4–A7 + subcoeruleus group in mammals. Their developmental characteristics show the largest similarities with the subcoeruleus group of birds and mammals, although some features are shared with developing A6 (locus coeruleus) neurons. Catecholaminergic neurons never appear during development in the ventral mesencephalon of the three-spined stickleback. A group of large dopaminergic neurons that accompany the cerebrospinal fluid (CSF)-contacting neurons follows the border between the hypothalamus and the ventral thalamus into the caudal hypothalamus, where they are continuous with the dopaminergic neurons in the posterior tuberculum. They are thus topologically comparable with the dopaminergic neurons of the zona incerta in mammals. The dopaminergic CSF-contacting neurons that line the median, lateral and posterior recesses of the third ventricle do not contain tyrosine hydroxylase-immunoreactivity at any developmental stage. This indicates that they take up and accumulate exogenous dopamine or l-dihydroxyphenylalanine, and do not synthesize dopamine from tyrosine at any developmental stage. Tyrosine hydroxylase-immunoreactive neurons appear in the pineal organ on the day of hatching (120 h post-fertilization). They were still observed in 240-h-old larvae, but are absent in the pineal organ of adult sticklebacks. The initial appearance and subsequent differentiation of catecholaminergic neurons in the stickle-back embryo follow essentially the same spatial and temporal pattern as in amphibian, avian and mammalian embryos. This observation supports the hypothesis that morphologically, topologically and chemically similar monoaminergic neurons in different vertebrate classes are homologous.