Straightforward upriver migration to spawning sites by chum salmon Oncorhynchus keta homing to coastal short rivers in the Sanriku region.

In chum salmon (Oncorhynchus keta) homed to the Sanriku region, Japan, most of the fish are matured in bays and spawn near river mouths in coastal short rivers; therefore, their upriver migration is extremely short, but their behavioural characteristics have remained unknown. Upriver migration in the Otsuchi River, a typical coastal river, was evaluated from behavioural and physiological aspects. Homing salmon tracked in Otsuchi Bay held in the inner bay for less than 1 day to more than 10 days before river entry. The varied holding duration was negatively correlated with plasma 17α, 20ß-dihydroxy-4-pregnen-3-one (DHP) concentration, an indicator of maturation. After river entry, however, most fish were captured in weirs near the river mouths within 2 days regardless of the DHP concentration. Of the 34 fish released in the river, on the contrary, eighteen and five fish were seen next day in the main spawning sites located at c. 1.5 km upstream and in the branch creek, respectively, and 85% of the fish held position there until their death. The mean survival time of released fish was 5.8 days. Plasma DHP level suggested that preparations for spawning were already completed at the timing of the release. Taken together, homing salmon completed spawning preparation in the bay, and then they moved to their spawning sites immediately after river entry and spawned there during their short remaining life. This upriver migration contrasts with those of other populations, such as early migrants and long river migrants, whose maturation is completed during upriver migration.

A comparison of avoidance to acoustic stimuli in fish with different auditory capabilities: juvenile chum salmon (Oncorhynchus keta) and common carp (Cyprinus carpio).

This study compared the repulsive effects of sound playbacks of intermittent 30, 150, 300, 600 and 900 Hz tones on two fish with different auditory capabilities: juvenile chum salmon (Oncorhynchus keta) and common carp (Cyprinus carpio). When 150 and 300 Hz tones were emitted from an underwater speaker, O. keta exhibited a moderate repulse reaction. Conversely, C. carpio exhibited a moderate repulse reaction to a tone with a frequency of 30 Hz, which indicates that a low-frequency component in complex broadband sound may be important for inducing a repulse reaction in cyprinids.

Chum salmon migrating upriver adjust to environmental temperatures through metabolic compensation.

Ectotherms adjust their thermal performance to various thermal ranges by altering their metabolic rates. These metabolic adjustments involve plastic and/or genetic traits and pathways depend on species-specific ecological contexts. Chum salmon (Oncorhynchus keta) are ecologically unique among the Pacific salmonids as early-run and late-run populations are commonly observed in every part of their range. In the Sanriku coastal area, Japan, early-run adults experience high water temperatures (12-24°C) during their migration, compared with those of the late-run adults (4-15°C), suggesting that the two populations might have different thermal performance. Here, we found population-specific differences in the thermal sensitivities of metabolic rates [resting metabolic rate, RMR, and maximum (aerobic) metabolic rate, MMR] and critical temperature maxima. Using these parameters, we estimated thermal performance curves of absolute aerobic scope (AAS). The populations had different thermal performance curves of AAS, and in both populations high values of AAS were maintained throughout the range of ecologically relevant temperatures. However, the populations did not vary substantially in the peak (AAS at optimal temperature, ToptAAS) or breadth (width of sub-optimal temperature range) of the performance curves. The AAS curve of early-run fish was shifted approximately 3°C higher than that of late-run fish. Furthermore, when the data for RMR and MMR were aligned to the thermal differences from ToptAAS, it became clear that the populations did not differ in the temperature dependence of their metabolic traits. Our results indicate that chum salmon thermally accommodate through compensatory alterations in metabolic rates. Our results imply that metabolic plasticity and/or the effect of genetic variance on plasticity might play a pivotal role in their thermal accommodation.

Spreading of River Water Guides Migratory Behavior of Homing Chum Salmon Oncorhynchus keta in Otsuchi Bay, a Narrow Inlet with Multiple River Flows.

The Sanriku-ria coast of Japan, a homing area for chum salmon, Oncorhynchus keta, is characterized by a large number of small closed bays into which one or multiple short rivers flow. The present behavioral investigation of chum salmon in this region was designed to gain deeper insight into the migration of chum salmon to their natal rivers. Eighty-three fish caught at the middle part of Otsuchi Bay were tracked using an acoustic transmitter in the narrow inlet into which flow three rivers: the Otsuchi, Koduchi, and Unosumai. The majority of 18 fish that entered the Unosumai River, which flows into the southwest side of the bay, directly approached the river along the southern coast. More than half of fish that entered the Otsuchi and Koduchi Rivers, which flow into the northwest side, also migrated into the inner bay via the southerly route, and then entered these rivers frequently after passing the mouth of the Unosumai River. In the inner bay, the salinity of sea surface water suggested that water from the three rivers circulates in a counterclockwise direction at a depth of less than 1.0 m, flowing eastwardly along the southern coast. The observed migratory paths of homing salmon in Otsuchi Bay thus correspond well with the counterflow of surface river water in the bay. The present results suggest that homing migration of salmon in the Sanriku narrow inlet is guided by natal river flows.

Enhanced osmoregulatory ability marks the smoltification period in developing chum salmon (Oncorhynchus keta).

The freshwater (FW) life of chum salmon is short, as they migrate to the ocean soon after emergence from the substrate gravel of natal waters. The alevins achieve seawater (SW) acclimating ability at an early developmental stage and the details of smoltification are not clear. We examined the stage-dependent SW acclimating ability in chum salmon alevins and found a sharp increase in SW tolerance during development that resembles the physiological parr-smolt transformation seen in other salmonids. Perturbation of plasma Na+ after SW exposure was prominent from the hatched embryo stage to emerged alevins, but the plasma Na+ became highly stable and more resistant to perturbation soon after complete absorption of yolk. Marker gene expression for SW-ionocytes including Na/K-ATPase (NKA α1b), Na-K-Cl cotransporter 1a (NKCC1a), Na/H exchanger 3a (NHE3a), cystic fibrosis transmembrane conductance regulators (CFTR I and CFTR II) were all upregulated profoundly at the same stage when the alevins were challenged by SW, suggesting that the stability of plasma Na+ concentration was partly a result of elevated osmoregulatory capability. FW-ionocyte markers including NKA α1a and NHE3b were consistently downregulated independent of stage by SW exposure, suggesting that embryos at all stages respond to salinity challenge, but the increase in SW osmoregulatory capability is restricted to the developmental stage after emergence. We propose that the "smoltification period" is condensed and integrated into the early development of chum salmon, and our results can be extrapolated to the future studies on hormonal controls and developmental triggers for smoltification in salmonids.

Molecular characterization and gene expression of syntaxin-1 and VAMP2 in the olfactory organ and brain during both seaward and homeward migrations of chum salmon, Oncorhynchus keta.

Anadromous Pacific salmon (Genus Oncorhynchus) imprint odorants from their natal streams during their seaward migration, and adult salmon use olfaction to identify their natal streams during their homeward migration. However, little is known about the molecular mechanisms of olfactory imprinting in the salmon nervous system. Our previous study suggested that the snap25s gene (encoding a soluble N-ethylmaleimide-sensitive factor attachment protein receptor [SNARE] protein) is involved in pre-synaptic functions for olfactory imprinting and/or olfactory memory retrieval in chum salmon (O. keta). In this study, the expression of other SNARE proteins was analyzed in chum salmon brains. Three cDNAs, encoding salmon SNARE proteins (STX-1a, STX-1b, and VAMP2), were isolated and sequenced, which are well-conserved among vertebrates. Quantitative PCR detected the expression of stx1s and vamp2 in all regions of the brain, and especially highly in the olfactory bulb (OB) and telencephalon. The expression levels of snares in the olfactory rosette (OR) were higher during seaward migration than in adult life stages, subsequently vamp2 in the OB and telencephalon increased during seaward migration, corresponding well with development of the olfactory nervous system. Both stx1s in the OB and stx1b in the telencephalon were elevated in the seaward period, whereas stx1a in the telencephalon increased continuously until the feeding period. Both stx1s in the telencephalon increased in the last phase of upriver migration, possibly related to the retrieval of imprinted memory. Our results indicated the involvement and distinct roles of upregulated snares in synaptic plasticity for olfactory imprinting and/or olfactory memory retrieval in Pacific salmon.

The behavioural homing response of adult chum salmon Oncorhynchus keta to amino-acid profiles.

Adult chum salmon Oncorhynchus keta homing behaviour in a two-choice test tank (Y-maze) was monitored using a passive integrated transponder (PIT)-tag system in response to river-specific dissolved free amino-acid (DFAA) profiles and revealed that the majority of O. keta showed a preference for artificial natal-stream water and tended to stay in this maze arm for a longer period; natal-stream water was chosen over a nearby tributary's water, but not when the O. keta were presented with a non-tributary water. The results demonstrate the ability of O. keta to discriminate artificial stream waters containing natural levels of DFAA.

Temperature seasonality during fry out-migration influences the survival of hatchery-reared chum salmon Oncorhynchus keta.

Among years, fry-to-adult survival of hatchery-reared chum salmon Oncorhynchus keta was positively correlated with the length (in days) of the fry out-migration period with temperatures suitable for migration. Furthermore, survival decreased with increasing difference in mean temperature between May and June. Thus, prolonged out-migration periods increased the probability of survival from fry to adult, lending support to the hypothesis that long migration periods decrease the risk of mortality (bet-hedging), and increase the probability of migration when environmental conditions in fresh water and the ocean are suitable (match-mismatch).

Circulating insulin-like growth factor I in juvenile chum salmon: relationship with growth rate and changes during downstream and coastal migration in northeastern Hokkaido, Japan.

Chum salmon (Oncorhynchus keta) migrate to the ocean in their first spring, and growth during early marine life is critical for survival. We examined the validity of circulating IGF-I and muscle RNA/DNA ratio as indices of growth rate using individually tagged juvenile chum salmon fed or fasted for 10 days. Serum IGF-I level was highly, positively correlated with individual growth rate. Muscle RNA/DNA ratio also showed a positive correlation, but its relation was not as high as that of IGF-I. We next measured these physiological parameters in chum salmon juveniles caught at river, estuary, port and nearshore of the northeastern Hokkaido, Japan, from May to June in 2013 and 2014, respectively. In both years, there was a trend that serum IGF-I levels were high in nearshore fish and low in river/estuarine fish in June. In contrast, muscle RNA/DNA ratio showed no clear temporal and spatial patterns. The present study shows that circulating IGF-I can be used as a growth index in juvenile chum salmon. Monitoring growth status using serum IGF-I suggests that growth of juvenile chum salmon in the survey area was activated when they left the coast.

[Downstream migration, behavior, and distribution of fish fry in the lower reaches of the Ozernaya River (southwestern Kamchatka)].

Fry of five species of salmonids are found in the lower reaches of the Ozernaya River. The most abundant are chum salmon and pink salmon which compose the bulk of fry which migrate downstream from the river to the sea. The dates and duration of migration of particular species differed according to the specific traits of their biology. Pink salmon is characterized by a simple migration strategy: it migrated downstream in a short time after emergence from theground. Chum salmon has two strategies of downstream migration: some fry start migration soon after emergence, and others remained in the river for several weeks. Downstream migration of pink salmon occurred mainly at night in contrast to that of chum salmon, over 24 h, the part of daytime increased with growth, of the fish. Migration of pink salmon was passive. Passive migration of chum salmon changed into active-passive with growth of the fish. The ratio of fish in the inshore zone and in the current was different in the course of 24 h. The number of fish in the inshore zone decreased in the period of intensive downstream migration.

Olfactory homing of chum salmon to stable compositions of amino acids in natal stream water.

Many attempts have been made to identify natal stream odors for salmon olfactory homing. It has recently been hypothesized that odors are dissolved free amino acids; however, it is unknown whether these odors change on a seasonal or annual basis. We analyzed dissolved free amino acid (DFAA) concentration and composition of water from the Teshio River in Hokkaido, Japan, where chum salmon (Oncorhynchus keta) returned for spawning, during juvenile downstream migration in spring and adult upstream migration in autumn with a 4-year difference. Among the 19 amino acids found in the Teshio River water, DFAA concentrations fluctuated largely, but 5-7 stable DFAA compositions (mole %) were found between the spring and autumn samples over a 4-year span. Two kinds of artificial stream water (ASW) were prepared using the same DFAA concentration in the Teshio River during the time of juvenile imprinting in spring (jASW) and adult homing in autumn (aASW), after a 4-year period. In behavioral experiments of upstream selective movement in a 2choice test tank, 4-year-old mature male chum salmon captured in the Teshio River showed significant preference for either jASW or aASW when compared to control water, but did not show any preference with respect to jASW or aASW. In electro-olfactogram experiments, adults were able to discriminate between jASW and aASW. Our findings demonstrate that the long-term stability of the DFAA compositions in natal streams may be crucial for olfactory homing in chum salmon.

Chum salmon Oncorhynchus keta respond to moonlight during homeward migrations.

The swimming depth of chum salmon Oncorhynchus keta equipped with archival tags was investigated off the Pacific Ocean coast of Hokkaido and North Honshu, Japan. As shown from movements of the fish with disc tags, O. keta swam at shallower depths during the full-moon phase than in the other phases and their swimming speed during this phase was faster compared to other phases. In addition, the circadian rhythm suggests a biological clock. These observations are all consistent with the view that O. keta make use of moonlight in order to navigate at night-time during homeward migration.

EMG telemetry studies on upstream migration of chum salmon in the Toyohira River, Hokkaido, Japan.

The movements of 28 adult chum salmon, Oncorhynchus keta (Walbaum) tagged with electromyogram (EMG) transmitters were tracked along the Toyohira river, Hokkaido, Japan, in October of 2007 and 2008 to investigate and evaluate the upstream migratory behavior through the protection bed and fishway of ground sills. The approach time of fish that ascended successfully through the protection bed and fishway was shorter than that of unsuccessful fish. The unsuccessful fish were observed to swim in currents with high water velocity and shallow water depth at swimming speeds that exceeded their critical swimming speed (U (crit)) during the approach to these structures. In consequence, unsuccessful fish frequently alternated between burst and maximum sustained speeds without ever ascending the fishway, and eventually became exhausted. It is important that fishway are constructed to enable chum salmon to find a passage way easily, so that they can migrate upstream rapidly without wasting excessive energy.

Molecular characterization of transient receptor potential vanilloid 4 (TRPV4) gene transcript variant mRNA of chum salmon Oncorhynchus keta in response to salinity or temperature changes.

Transient receptor potential vanilloid 4 (TRPV4) is an osmosensory cation channel that respond to an increase in cell volume and participates in various physiological functions. Among organisms in aquatic environments, euryhaline teleost is are suitable experimental models to study ion channel proteins related to physiological functions involving osmosensing. Among the studies of various regulatory molecules that mediate osmotic regulation in fish, however, information is lacking, particularly on the TRP family. This study investigated the structural characteristics of theTRPV4 gene of chum salmon (Oncorhynchus keta) and their responses to changes in salinity and temperature. Interestingly, TRPV4 generates transcript variants of the intron-retention form through alternative splicing, resulting in a frameshift leading to the generation of transcripts of different structures. In particular, TRPV4 x1 and TRPV x2 mRNAs were predominant in the gill and skin including at the lateral line. The expression levels of chum salmon TRPV4 x1 were significantly increased with increase in salinity and temperature, whereas TRPV4 x2 mainly responded to temperature decrease. Overall, these results demonstrate for the first time the effects of salinity and temperature on the expression of two salmonid TRPV4 transcript variants, suggesting their contribution to the regulation of hydromineral balance.

Expression of GnRH genes is elevated in discrete brain loci of chum salmon before initiation of homing behavior and during spawning migration.

Our previous studies suggested the importance of gonadotropin-releasing hormones (GnRHs) for initiation of spawning migration of chum salmon, although supporting evidence had been not available from oceanic fish. In farmed masu salmon, the amounts of salmon GnRH (sGnRH) mRNAs in the forebrain increased in the pre-pubertal stage from winter through spring, followed by a decrease toward summer. We thus hypothesized that gene expression for GnRHs in oceanic chum salmon changes similarly, and examined this hypothesis using brain samples from winter chum salmon in the Gulf of Alaska and summer fish in the Bering Sea. They were classified into sexually immature and maturing adults, which had maturing gonads and left the Bering Sea for the natal river by the end of summer. The absolute amounts of GnRH mRNAs were determined by real-time PCRs. The amounts of sGnRH mRNA in the maturing winter adults were significantly larger than those in the maturing summer adults. The amounts of sGnRH and chicken GnRH mRNAs then peaked during upstream migration from the coast to the natal hatchery. Such changes were observed in various brain loci including the olfactory bulb, terminal nerve, ventral telencephalon, nucleus preopticus parvocellularis anterioris, nucleus preopticus magnocellularis and midbrain tegmentum. These results suggest that sGnRH neurons change their activity for gonadal maturation prior to initiation of homing behavior from the Bering Sea. The present study provides the first evidence to support a possible involvement of neuropeptides in the onset of spawning migration.

Changes in the plasma levels of insulin-like growth factor-I from the onset of spawning migration through upstream migration in chum salmon.

An increase in activity of the pituitary-gonadal axis (PG-axis) and gonadal development are essential for the onset of spawning migration of chum salmon from the Bering Sea. In the Bering Sea, fish with larger body sizes initiated gonadal development and commenced spawning migration to the natal river by the end of summer. We thus hypothesized that insulin-like growth factor-I (IGF-I), a somatotropic signal that interacts with the PG-axis, can be one of such factors responsible for the onset of migration, and examined changes in plasma levels and hepatic expression of IGF-I gene in oceanic and homing chum salmon in 2001-2003. The plasma IGF-I levels and corresponding body sizes in maturing adults, which had developing gonads, were significantly higher than those in immature fish in all years examined. Such increase in the plasma IGF-I levels in maturing fish was observed even in the Gulf of Alaska during February 2006, while coincident increase was not observed in the hepatic amounts of IGF-I mRNA. In autumn, the plasma IGF-I levels in homing adults decreased during upstream migration in the Ishikari River-Ishikari bay water system in Hokkaido, Japan. In conclusion, the plasma IGF-I levels increased with gonadal development when chum salmon migrated from the winter Gulf of Alaska to the summer Bering Sea. Circulating IGF-I may interact with the PG-axis and promote gonadal development that is inseparable from the onset of spawning migration. Circulating IGF-I levels were thereafter lowered in accordance with final maturation during upstream migration in the breeding season.

Changes in gene expression for GH/PRL/SL family hormones in the pituitaries of homing chum salmon during ocean migration through upstream migration.

Gene expression for growth hormone (GH)/prolactin (PRL)/somatolactin (SL) family hormones in the pituitaries of homing chum salmon were examined, because gene expression for these hormones during ocean-migrating phases remains unclear. Fish were collected in the winter Gulf of Alaska, the summer Bering Sea and along homing pathway in the Ishikari River-Ishikari Bay water system in Hokkaido, Japan in autumn. The oceanic fish included maturing adults, which had developing gonads and left the Bering Sea for the natal river by the end of summer. The absolute amounts of GH, PRL and SL mRNAs in the pituitaries of the maturing adults in the summer Bering Sea were 5- to 20-fold those in the winter Gulf of Alaska. The amount of GH mRNA in the homing adults at the coastal seawater (SW) areas was smaller than that in the Bering fish, while the amount of PRL mRNA remained at the higher level until fish arrived at the Ishikari River. The gill Na(+),K(+)-ATPase activity in the coastal SW fish and the plasma Na(+) levels in the brackish water fish at the estuary were lowered to the levels that were comparable to those in the fresh water (FW) fish. In conclusion, gene expression for GH, PRL and SL was elevated in the pituitaries of chum salmon before initiation of homing behavior from the summer Bering Sea. Gene expression for GH is thereafter lowered coincidently with malfunction of SW adaptability in the breeding season, while gene expression for PRL is maintained high until forthcoming FW adaptation.

Comparative transcriptome profiling of selected osmotic regulatory proteins in the gill during seawater acclimation of chum salmon (Oncorhynchus keta) fry.

Salmonid fishes, chum salmon (Oncorhynchus keta) have the developed adaptive strategy to withstand wide salinity changes from the early life stage. This study investigated gene expression patterns of cell membrane proteins in the gill of chum salmon fry on the transcriptome level by tracking the salinity acclimation of the fish in changing environments ranging from freshwater (0 ppt) to brackish water (17.5 ppt) to seawater (35 ppt). Using GO analysis of DEGs, the known osmoregulatory genes and their functional groups such as ion transport, transmembrane transporter activity and metal ion binding were identified. The expression patterns of membrane protein genes, including pump-mediated protein (NKA, CFTR), carrier-mediated protein (NKCC, NHE3) and channel-mediated protein (AQP) were similar to those of other salmonid fishes in the smolt or adult stages. Based on the protein-protein interaction analysis between transmembrane proteins and other related genes, we identified osmotic-related genes expressed with salinity changes and analyzed their expression patterns. The findings of this study may facilitate the disentangling of the genetic basis of chum salmon and better able an understanding of the osmophysiology of the species.

Behavioral responses by migratory chum salmon to amino acids in natal stream water.

We propose that amino acids in natal stream water have Important roles in Pacific salmon homing. This study hypothesized that amino acids found in natal stream water have a role in the ability of mature male chum salmon (Oncorhynchus keta) to home to the Osaru River (OR), Hokkaido, Japan. Behavioral experiments were conducted in a two-choice test tank using various combinations of control water (natural Toya Lake water; NLW and three artificial stream waters using amino acids: 1) artificial OR water (AOR); 2) AOR without L-glutamic acid, the major amino acid in OR water (AOR-E); and 3) artificial water matching another stream (ALS) that had much higher amino acid concentrations than OR. In behavioral tests, the fish did not select between AOR and AOR-E, but still chose AOR over NLW, AOR-E over NLW, and AOR over ALS. These results suggest that migratory male chum salmon respond to amino acid mixtures in their natal stream water and appear to be affected by multiple amino acids.

Ontogenic change in tissue osmolality and developmental sequence of mitochondria-rich cells in Mozambique tilapia developing in freshwater.

We investigated a change in tissue fluid osmolality and developmental sequences of mitochondria-rich (MR) cells during embryonic and larval stages of Mozambique tilapia, Oreochromis mossambicus, developing in freshwater. Tissue osmolality, representing body fluid osmolality, ranged from 300 to 370 mOsm/kg during embryonic and larval stages. This suggests that tilapia embryos and larvae are also able to regulate body fluid osmolality to some extent, although the levels are somewhat higher and fluctuate more greatly in embryos and larvae than in adults. Na(+)/K(+)-ATPase-immunoreactive MR cells were first detected in the yolk-sac membrane 3 days before hatching (day -3), followed by their appearance in the body skin on day -2. Subsequently, MR cells in both the yolk-sac membrane and body skin increased in number, and most densely observed on days -1 and 0. Whereas yolk-sac and skin MR cells decreased after hatching, MR cells in turn started developing in the gills after hatching. Thus, the principal site for MR cell distribution shifted from the yolk-sac membrane and body skin during embryonic stages to the gills during larval stages, and tilapia could maintain continuously their ion balance through those MR cells during early life stages.