Olfactory Imprinting of Amino Acids in Lacustrine Sockeye Salmon

Juvenile salmon have an olfactory ability to imprint their natal stream odors, but neither the odor properties of natal stream water nor the imprinting timing and duration have been clarified as yet. Here we show, using electrophysiological and behavioral experiments, that one-year-old lacustrine sockeye salmon (Oncorhynchus nerka) can be imprinted around the stage of parr-smolt transformation (PST) by a single amino acid, 1 µM L-proline (Pro), or L-glutamic acid (Glu). We also show by real-time PCR that changes occur in mRNA levels of the salmon olfactory imprinting-related gene (SOIG) around PST. The electro-olfactogram (EOG) responses of test fish exposed to Pro in March (before PST) and April–June (during PST) for 2 weeks were significantly (1.7-fold) greater than those of non-exposed control fish, but not those of test fish exposed in July (after PST). When Pro and control water were added to the water inlets of a two-choice test tank during the spawning season 2 years after the test water exposure, 80% of maturing and matured test fish exposed before and during PST showed a preference for Pro, whereas those exposed after PST did not. The EOG response of test fish exposed to Pro or Glu for 1 hour, 6 hours, 1 day, 7 days, or 14 days in May revealed that only the response after 14 days of exposure was significantly (1.8-fold) greater than the control. The expression levels of SOIG mRNA increased before and during PST, and decreased after PST. We conclude that one-year-old lacustrine sockeye salmon can be imprinted by a single amino acid before and during PST, and that imprinting requires exposure for at least 14 days.     

Olfactory gene expression in migrating adult sockeye salmon Oncorhynchus nerka

Expression of 12 olfactory genes was analysed in adult sockeye salmon Oncorhynchus nerka nearing spawning grounds and O. nerka that had strayed from their natal migration route. Variation was found in six of these genes, all of which were olfc olfactory receptors and had lower expression levels in salmon nearing spawning grounds. The results may reflect decreased sensitivity to natal water olfactory cues as these fish are no longer seeking the correct migratory route. The expression of olfactory genes during the olfactory‐mediated spawning migration of Pacific salmon Oncorhynchus spp. is largely unexplored and these findings demonstrate a link between migratory behaviours and olfactory plasticity that provides a basis for future molecular research on salmon homing.     

Lacustrine sockeye salmon return straight to their natal area from open water using both visual and olfactory cues

Mechanisms of the amazing ability of salmon to migrate a long distance from open water to natal streams for spawning are still unknown. Lacustrine sockeye salmon (Oncorhynchus nerka) in Lake Toya offers an excellent model system for studying the orientation mechanism in open water, because mature fish return to the natal area with a high degree of accuracy. First we examined the percentage of fish returning to the natal area after they were released 7 km south of the natal area. Forty percent of control male mature fish and 25% of the fish blinded by injection of a mixture of carbon toner and corn oil into the eyeball were captured in the natal area within 5 days. Forty-four percent of fish with brass rings (control) and 31% of fish with NdFe magnetic rings which interfere with the magnetic cue were captured in the natal area within 3 days. These experiments suggested that, although the number of blinded fish captured in the natal area was less than that of the controls, the difference was not statistically significant. In the fish captured in the natal area within 3 or 5 days, fish which found the natal area using their olfactory cue after random swimming for a long time and returned to that area may be included. Hence we tracked fish telemetrically using an ultrasonic tracking system, and found that mature males released at a long distance (3.6 or 6.8 km) from the natal area swam straight to the vicinity of the natal area. Interference of the magnetic cue by the attachment of a magnetic ring did not affect their direct return. Blockage of the visual cue caused them to move randomly. These data suggest that lacustrine sockeye salmon return straight to the vicinity of the natal area using their visual cue and finally reach the exact homing point using their olfactory cue.      

Gene expression of neuronal soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex components in the olfactory organ and brain during seaward and homeward migration by pink salmon (Oncorhynchus gorbuscha)

The expression of synaptic vesicle exocytosis-regulator SNARE complex component genes (snap25, stx1 and vamp2) was examined in the olfactory nervous system during seaward and homeward migration by pink salmon (Oncorhynchus gorbuscha). The expression levels of snares in the olfactory organ were higher in seaward fry than in feeding and homeward adults, reflecting the development of the olfactory nervous system. The expression of snap25a, b and stx1a was upregulated or stable in the adult olfactory bulb and telencephalon. This upregulated expression suggested alterations in olfactory neuronal plasticity that may be related to the discrimination of natal rivers. The expression of stx1b was downregulated in the adult olfactory bulb, but remained stable in the adult telencephalon. The expression of vamp2 was initially strong in seaward fry, but was downregulated in adults in both the olfactory bulb and telencephalon. Pink salmon has the lowest diversity of maturation age, the largest population, and the most evolutional position in Pacific salmon (genus Oncorhynchus). The expression of snares in the olfactory center of pink salmon reflected the timing of sexual maturation and homeward migration. The present results and our previous studies indicate that snares show distinct expression patterns between two salmon species that depend on physiological and ecological features of migration.     

Development of the olfactory system of the white sucker,Catostomus commersoni,in relation to imprinting and homing: a comparison to the salmonid model

Synopsis White suckers,Catastomus commersoni, use olfactory cues to return to the same spawning stream year after year. If we assume that they follow a model similar to the well-known salmon model, olfactory imprinting must occur very early in their development. We describe the time of migration from the nursery stream in relation to the development of the white sucker olfactory system to determine if the requisite anatomical structures are present which would permit imprinting. At hatching the olfactory placode is present and beginning to differentiate, the lumen of the olfactory capsule is starting to form, and the olfactory tract projects into the telencephalon. Larvae migrate approximately 2 weeks later or at a size of 14 mm TL. At this time olfactory cilia are present, the olfactory tract is robust and the telencephalon is beginning to differentiate. Therefore, it appears that the fundamental neural structures necessary for imprinting are present. A comparison with salmon, however, clearly demonstrates that the white sucker olfactory apparatus is not as well developed as that of salmon at time of migration. This raises the question of the ability of white suckers to imprint in the same manner as salmon and whether the salmonid model is applicable to white suckers. Alternative imprinting hypotheses are discussed.     

Sensory mechanisms of natal stream imprinting and homing in Oncorhynchus spp.

Juvenile Oncorhynchus spp. can memorise their natal stream during downstream migration; juveniles migrate to feed during their growth phase and then they migrate long distances from their feeding habitat to their natal stream to reproduce as adults. Two different sensory mechanisms, olfaction and navigation, are involved in the imprinting and homing processes during short-distance migration within the natal stream and long-distance migration in open water, respectively. Here, olfactory functions are reviewed from both neurophysiological studies on the olfactory discrimination ability of natal stream odours and neuroendocrinological studies on the hormonal controlling mechanisms of olfactory memory formation and retrieval in the brain. These studies revealed that the long-term stability of dissolved free amino-acid composition in the natal stream is crucial for olfactory imprinting and homing. Additionally, the brain–pituitary–thyroid and brain–pituitary–gonadal hormones play important roles in olfactory memory formation and retrieval, respectively. Navigation functions were reviewed from physiological biotelemetry techniques with sensory interference experiments during the homing migration of anadromous and lacustrine Oncorhynchus spp. The experiments demonstrated that Oncorhynchus spp. used compass navigation mechanisms in the open water. These findings are discussed in relation to the sensory mechanisms involved in natal stream imprinting and homing in Oncorhynchus spp.     

Physiological mechanisms of imprinting and homing migration in Pacific salmon Oncorhynchus spp

After several years of feeding at sea, salmonids have an amazing ability to migrate long distances from the open ocean to their natal stream to spawn. Three different research approaches from behavioural to molecular biological studies have been used to elucidate the physiological mechanisms underpinning salmonid imprinting and homing migration. The study was based on four anadromous Pacific salmon Oncorhynchus spp., pink salmon Oncorhynchus gorbuscha, chum salmon Oncorhynchus keta, sockeye salmon Oncorhynchus nerka and masu salmon Oncorhynchus masou, migrating from the North Pacific Ocean to the coast of Hokkaido, Japan, as well as lacustrine O. nerka and O. masou in Lake Toya, Hokkaido, where the lake serves as the model oceanic system. Behavioural studies using biotelemetry techniques showed swimming profiles from the Bering Sea to the coast of Hokkaido in O. keta as well as homing behaviours of lacustrine O. nerka and O. masou in Lake Toya. Endocrinological studies on hormone profiles in the brain-pituitary-gonad axis of O. keta, and lacustrine O. nerka identified the hormonal changes during homing migration. Neurophysiological studies revealed crucial roles of olfactory functions on imprinting and homing during downstream and upstream migration, respectively. These findings are discussed in relation to the physiological mechanisms of imprinting and homing migration in anadromous and lacustrine salmonids.     

Ontogenetic shifts in olfactory rosette morphology of the sockeye salmon,Oncorhynchus nerka

Sockeye salmon, Oncorhynchus nerka, are anadromous, semelparous fish that breed in freshwater—typically in streams, and juveniles in most populations feed in lakes for 1 or 2 years, then migrate to sea to feed for 2 or 3 additional years, before returning to their natal sites to spawn and die. This species undergoes important changes in behavior, habitat, and morphology through these multiple life history stages. However, the sensory systems that mediate these migratory patterns are not fully understood, and few studies have explored changes in sensory function and specialization throughout ontogeny. This study investigates changes in the olfactory rosette of sockeye salmon across four different life stages (fry, parr, smolt, and adult). Development of the olfactory rosette was assessed by comparing total rosette size (RS), lamellae number, and lamellae complexity from scanning electron microscopy images across life stages, as a proxy for olfactory capacity. Olfactory RS increased linearly with lamellae number and body size (p < .001). The complexity of the rosette, including the distribution of sensory and nonsensory epithelia and the appearance of secondary lamellar folding, varied between fry and adult life stages. These differences in epithelial structure may indicate variation in odor-processing capacity between juveniles imprinting on their natal stream and adults using those odor memories in the final stages of homing to natal breeding sites. These findings improve our understanding of the development of the olfactory system throughout life in this species, highlighting that ontogenetic shifts in behavior and habitat may coincide with shifts in nervous system development.     

Two differing salmon GnRH precursor mRNAs are co-expressed in the brain of sockeye salmon (Oncorhynchus nerka)

The localization of two salmon-type gonadotropin-releasing hormone (sGnRH) precursors, pro-sGnRH-I (short type) and pro-sGnRH-II (long type), was investigated by using in situ hybridization techniques in the brain of the landlocked sockeye salmon, Oncorhynchus nerka. We used 30-mer oligonucleotide probes complementary to pro-sGnRH-I and pro-sGnRH-II cDNA. No significant differences were observed in the localization of sGnRH neurons expressing pro-sGnRH-I and pro-sGnRH-II mRNAs; both were expressed in the olfactory nerve, the olfactory bulbs, the regions between the olfactory bulb and telencephalon, the ventral telencephalon, the preoptic area, and the hypothalamus. Almost all sGnRH neurons examined co-expressed both precursors. The expression of two sGnRH precursors in the same neuron and the wide distribution of such neurons in the brain suggest that there are no functional differences between the two precursors.     

Pacific salmon in hot water: applying aerobic scope models and biotelemetry to predict the success of spawning migrations

Concern over global climate change is widespread, but quantifying relationships between temperature change and animal fitness has been a challenge for scientists. Our approach to this challenge was to study migratory Pacific salmon (Oncorhynchus spp.), fish whose lifetime fitness hinges on a once-in-a-lifetime river migration to natal spawning grounds. Here, we suggest that their thermal optimum for aerobic scope is adaptive for river migration at the population level. We base this suggestion on several lines of evidence. The theoretical line of evidence comes from a direct association between the temperature optimum for aerobic metabolic scope and the temperatures historically experienced by three Fraser River salmon populations during their river migration. This close association was then used to predict that the occurrence of a period of anomalously high river temperatures in 2004 led to a complete collapse of aerobic scope during river migration for a portion of one of the sockeye salmon (Oncorhynchus nerka) populations. This prediction was corroborated with empirical data from our biotelemetry studies, which tracked the migration of individual sockeye salmon in the Fraser River and revealed that the success of river migration for the same sockeye population was temperature dependent. Therefore, we suggest that collapse of aerobic scope was an important mechanism to explain the high salmon mortality observed during their migration. Consequently, models based on thermal optima for aerobic scope for ectothermic animals should improve predictions of population fitness under future climate scenarios.     

Cortisol and Pacific Salmon: A New Look at the Role of Stress Hormones in Olfaction and Home-stream Migration

Pacific salmon (genus Oncorhynchus) exhibit an interesting anduncommon life-history pattern that combines semelparity, anadromy,and navigation (homing). During smoltification, young salmonimprint on the chemical composition of their natal stream water(the home-stream olfactory bouquet or "HSOB"); they then migrateto the ocean where they spend a few years feeding prior to migratingback to their natal freshwater stream to spawn. Upstream migrationis guided by the amazing ability to discriminate between thechemical compositions of different stream waters and thus identifyand travel to their home-stream. Pacific salmon demonstratemarked somatic and neural degeneration changes during home-streammigration and at the spawning grounds. The appearance of thesepathologies is correlated with a marked elevation in plasmacortisol levels. While the mechanisms of salmonid homing arenot completely understood, it is known that adult salmon continuouslyutilize two of their primary sensory systems, olfaction andvision, during homing. Olfaction is the primary sensory systeminvolved in freshwater homing and "HSOB" recognition, and willbe emphasized here. Previously, we proposed that the increasein plasma cortisol during Pacific salmon home-stream migrationis adaptive because it enhances the salmon's ability to recallthe imprinted memory of the "HSOB" (Carruth, 1998; Carruth etal., 2000b). Elevated plasma concentrations of cortisol couldprime the hippocampus or other olfactory regions of the brainto recall this memory and, therefore, aid in directing the fishto their natal stream. Thus, specific responses of salmon tostressors could enhance reproductive success.     

Rheoreaction type of early juveniles of sockeye salmon Oncorhynchus nerka in the Ozernaya River and Lake Kurilskoe

Rheoreaction of early juveniles of sockeye salmon Oncorhynchus nerka is experimentally investigated, including different spatial groups (denatant migrants, contranatant migrants, specimens living in bays and at closed river banks, juveniles from the coastal zone of the lake). Investigation is made in water of the river flowing from the lake and in water of the stream flowing into the lake. It is shown that the rheoreaction (the type of rheoreaction) is one of behavioral mechanisms performing feeding migration of early juveniles of sockeye salmon. Contranatant migrants that reached the feeding lake retain the positive type of rheoreaction, which favors the exit of juveniles from the zone of discharge exclusion and prevents the drift of juveniles to the river at night at poorer conditions for orientation. Juveniles placed into the water of the inflowing stream manifest a sharply negative rheoreaction type contributing to their rapid lakeward migration for feeding. In water of two steams (taking its beginning in the lake and flowing into it) such type of rheoreaction is observed which ensures migration of sockeye juveniles to their principal feeding water body-Lake Kurilskoe.     

Going with the Flow: Spatial Distributions of Juvenile Coho Salmon Track an Annually Shifting Mosaic of Water Temperature

A critical challenge for ecologists is to understand the functional significance of habitat heterogeneity and connectivity for mobile animals. Here, we explore how a thermo-regulating fish responds to annual variation in the spatial patterning of thermal and trophic resources. In a third-order stream in coastal Alaska, juvenile coho salmon forage on sockeye salmon eggs at night in cold water and then move to warmer water to increase their digestive capacity. We mapped the spatial distributions of water temperature, juvenile coho salmon, and spawning sockeye salmon across a 5-year period during which summer discharge varied by greater than fivefold. In low flow years, warm water (9–12°C) was only available in thalweg (that is, main-channel) habitat at least approximately 400 m upstream of the cooler habitat (3–7°C) where sockeye salmon spawned. In high flow years, the entire stream thalweg was isothermal at 7–8°C, but inundated off-channel areas generated warm habitats (9–12°C) laterally adjacent to the downstream regions where sockeye salmon spawned. The daytime spatial distribution of juvenile coho salmon shifted from headwater thalweg habitats in low flow years, to downstream off-channel habitats in high flow years. In all years, the majority of juvenile coho salmon sampled during the daytime were found in warm habitat units without sockeye salmon present, yet they exhibited diet contents comprised virtually entirely of sockeye salmon eggs. Thus, thermoregulatory movements by coho salmon were able to track an annually shifting mosaic of water temperature. Our results demonstrate how the spatial habitat heterogeneity and connectivity of intact floodplains can in turn buffer aquatic organisms from high levels of temporal variation in habitat conditions and resource abundance.     

Elevated CO2 impairs olfactory‐mediated neural and behavioral responses and gene expression in ocean‐phase coho salmon (Oncorhynchus kisutch)

Elevated concentrations of CO₂ in seawater can disrupt numerous sensory systems in marine fish. This is of particular concern for Pacific salmon because they rely on olfaction during all aspects of their life including during their homing migrations from the ocean back to their natal streams. We investigated the effects of elevated seawater CO₂ on coho salmon (Oncorhynchus kisutch) olfactory‐mediated behavior, neural signaling, and gene expression within the peripheral and central olfactory system. Ocean‐phase coho salmon were exposed to three levels of CO₂, ranging from those currently found in ambient marine water to projected future levels. Juvenile coho salmon exposed to elevated CO₂ levels for 2 weeks no longer avoided a skin extract odor that elicited avoidance responses in coho salmon maintained in ambient CO₂ seawater. Exposure to these elevated CO₂ levels did not alter odor signaling in the olfactory epithelium, but did induce significant changes in signaling within the olfactory bulb. RNA‐Seq analysis of olfactory tissues revealed extensive disruption in expression of genes involved in neuronal signaling within the olfactory bulb of salmon exposed to elevated CO₂, with lesser impacts on gene expression in the olfactory rosettes. The disruption in olfactory bulb gene pathways included genes associated with GABA signaling and maintenance of ion balance within bulbar neurons. Our results indicate that ocean‐phase coho salmon exposed to elevated CO₂ can experience significant behavioral impairments likely driven by alteration in higher‐order neural signal processing within the olfactory bulb. Our study demonstrates that anadromous fish such as salmon may share a sensitivity to rising CO₂ levels with obligate marine species suggesting a more wide‐scale ecological impact of ocean acidification.     

Environmental rearing conditions produce forebrain differences in wild Chinook salmon Oncorhynchus tshawytscha

Recent studies suggest that hatchery-reared fish can have smaller brain-to-body size ratios than wild fish. It is unclear, however, whether these differences are due to artificial selection or instead reflect differences in rearing environment during development. Here we explore how rearing conditions influence the development of two forebrain structures, the olfactory bulb and the telencephalon, in juvenile Chinook salmon (Oncorhynchus tshawytscha) spawned from wild-caught adults. First, we compared the sizes of the olfactory bulb and telencephalon between salmon reared in a wild stream vs. a conventional hatchery. We next compared the sizes of forebrain structures between fish reared in an enriched NATURES hatchery and fish reared in a conventional hatchery. All fish were size-matched and from the same genetic cohort. We found that olfactory bulb and telencephalon volumes relative to body size were significantly larger in wild fish compared to hatchery-reared fish. However, we found no differences between fish reared in enriched and conventional hatchery treatments. Our results suggest that significant differences in the volume of the olfactory bulb and telencephalon between hatchery and wild-reared fish can occur within a single generation.     

Elevation of gene expression for salmon gonadotropin-releasing hormone in discrete brain loci of prespawning chum salmon during upstream migration

Our previous studies suggested that salmon gonadotropin-releasing hormone (sGnRH) neurons regulate both final maturation and migratory behavior in homing salmonids. Activation of sGnRH neurons can occur during upstream migration. We therefore examined expression of genes encoding the precursors of sGnRH, sGnRH-I, and sGnRH-II, in discrete forebrain loci of prespawning chum salmon, Oncorhynchus keta. Fish were captured from 1997 through 1999 along their homing pathway: coastal areas, a midway of the river, 4 km downstream of the natal hatchery, and the hatchery. Amounts of sGnRH mRNAs in fresh frozen sections including the olfactory bulb (OB), terminal nerve (TN), ventral telencephalon (VT), nucleus preopticus parvocellularis anterioris (PPa), and nucleus preopticus magnocellularis (PM) were determined by quantitative real-time polymerase chain reactions. The amounts of sGnRH-II mRNA were higher than those of sGnRH-I mRNA, while they showed similar changes during upstream migration. In the OB and TN, the amounts of sGnRH mRNAs elevated from the coast to the natal hatchery. In the VT and PPa, they elevated along with the progress of final maturation. Such elevation was also observed in the rostroventral, middle, and dorsocaudal parts of the PM. The amounts of gonadotropin IIbeta and somatolactin mRNAs in the pituitary also increased consistently with the elevation of gene expression for sGnRH. These results, in combination with lines of previous evidence, indicate that sGnRH neurons are activated in almost all the forebrain loci during the last phases of spawning migration, resulting in coordination of final gonadal maturation and migratory behavior to the spawning ground.     

Non-direct homing behaviours by adult Chinook salmon in a large, multi-stock river system

Two non-direct homing behaviours, overshoot of natal tributaries and temporary non-natal tributary use, were evaluated for 5150 radio-tagged spring–summer Chinook salmon Oncorhynchus tshawytscha from 40 populations in the large Columbia River system. Over 7 years, 2–44% (mean = 15%) of individuals within each group temporarily entered presumed non-natal tributaries. In addition, many Chinook salmon from lower river tributaries initially travelled 3 to >250 km upstream in the main-stem river beyond confluences with presumed natal tributaries before returning to the natal sites ('overshoot'). Both overshoot and temporary tributary use behaviours declined exponentially with increasing distance from the natal tributary. Non-direct homing also increased later in the season as water temperatures rose and was associated with hatchery origin in some cases. The behaviours may reflect a mix of active searching for olfactory cues from natal sites, behavioural thermoregulation and orientation challenges in a large-river migration corridor transformed by dams and reservoirs. While anadromous salmonid homing is generally accurate and precise, these results indicate that route finding can be non-direct, potentially increasing energetic costs and harvest risks during migration.