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The ability to distinguish between different migratory behaviours (e.g., anadromy and potamodromy) in fish can provide important insights into the ecology, evolution, and conservation of many aquatic species. We present a simple stable carbon isotope (δ13C) approach for distinguishing between sockeye (anadromous ocean migrants) and kokanee (potamodromous freshwater residents), two migratory ecotypes of Oncorhynchus nerka (Salmonidae) that is applicable throughout most of their range across coastal regions of the North Pacific Ocean. Analyses of kokanee (n = 239) and sockeye (n = 417) from 87 sites spanning the North Pacific (Russia to California) show that anadromous and potamodromous ecotypes are broadly distinguishable on the basis of the δ13C values of their scale and bone collagen. We present three case studies demonstrating how this approach can address questions in archaeology, archival, and conservation research. Relative to conventional methods for determining migratory status, which typically apply chemical analyses to otoliths or involve genetic analyses of tissues, the δ13C approach outlined here has the benefit of being non-lethal (when applied to scales), cost-effective, widely available commercially, and should be much more broadly accessible for addressing archaeological questions since the recovery of otoliths at archaeological sites is rare.

To provide morphological bases for functional studies, the development of the photoreceptor cells in the retina of the cuttlefish Sepia esculenta from newly hatched juveniles to the adults was examined by light and electron microscopy. Four layers were distinguished in the retina: the rhabdomeric layer, subrhabdomeric layer, inner segmental layer, and plexiform layer. With the development of the retina, the structure of the plexiform layer becomes stronger. Morphometric data on the rhabdomeric microvilli from the newly hatched to the adult retinas are presented. The thickness of the rhabdomeric layer and the total surface area of the rhabdomeric microvilli present beneath a unit surface area of the retina show an exponential increase with developmental days; the diameter of the lens shows a linear increase with days. The results of our research show that the visual acuity and sensitivity of S. esculenta continuously increased with development.

Despite the importance of olfactory receptor neurons (ORNs) for homing migration, the expression of olfactory marker protein (OMP) is not well understood in ORNs of Pacific salmon (genus Oncorhynchus ). In this study, salmon OMP was characterized in the olfactory epithelia of lacustrine sockeye salmon ( O. nerka ) by molecular biological and histochemical techniques. Two cDNAs encoding salmon OMP were isolated and sequenced. These cDNAs both contained a coding region encoding 173 amino acid residues, and the molecular mass of the two proteins was calculated to be 19,581.17 and 19,387.11 Da, respectively. Both amino acid sequences showed marked homology (90%). The protein and nucleotide sequencing demonstrates the existence of high-level homology between salmon OMPs and those of other teleosts. By in situ hybridization using a digoxigenin-labeled salmon OMP cRNA probe, signals for salmon OMP mRNA were observed preferentially in the perinuclear regions of the ORNs. By immunohistochemistry using a specific antibody to salmon OMP, OMP-immunoreactivities were noted in the cytosol of those neurons. The present study is the first to describe cDNA cloning of OMP in salmon olfactory epithelium, and indicate that OMP is a useful molecular marker for the detection of the ORNs in Pacific salmon.

It is generally accepted that anadromous Pacific salmon (genus Oncorhynchus) imprint to odorants in their natal streams during their seaward migration and use olfaction to identify these during their homeward migration. Despite the importance of the olfactory organ during olfactory imprinting, the development of this structure is not well understood in Pacific salmon. Olfactory cues from the environment are relayed to the brain by the olfactory receptor neurons (ORNs) in the olfactory organ. Thus, we analyzed morphometric changes in olfactory lamellae of the peripheral olfactory organ and in the quantity of ORNs during life history from alevin to mature in chum salmon (Oncorhynchus keta). The number of lamellae increased markedly during early development, reached 18 lamellae per unilateral peripheral olfactory organ in young salmon with a 200 mm in body size, and maintained this lamellar complement after young period. The number of ORNs per olfactory organ was about 180,000 and 14.2 million cells in fry and mature salmon, respectively. The relationship between the body size (fork length) and number of ORNs therefore revealed an allometric association. Our results represent the first quantitative analysis of the number of ORNs in Pacific salmon and suggest that the number of ORNs is synchronized with the fork length throughout its life history.

Juveniles of chum salmon (Oncorhynchus keta) held at high population density were apparently smaller than those held at medium and low population densities. The effects of high population density on pituitary growth hormone (GH) cells in juvenile chum salmon were examined using immunocytochemical and in situ hybridization techniques. The ratio of GH-immunoreactive (ir) area to the whole pituitary was almost constant in all of the high, medium and low population density groups, although the number and sizes of GH-ir cells were decreased in the high population density group. Image-analysis of GH-ir cells indicated the presence of a population of heterogenous cells, in which medium or rather strongly stained smaller cells and as extreme weakly stained larger cells. The medium or rather strongly stained smaller cells predominated in the high population density group, while weakly stained larger cells in the low population density group. In situ hybridization study showed somewhat different distributions and intensities of hybridization signals for mRNAs encoding GH I and II precursors. The area showing signals for GH II mRNA in the high population density group was significantly smaller than those in the medium and low population density groups. In contrast, the sizes of areas showing signals for GH I mRNA did not differ among the groups, although the intensity was slightly higher in the high population density group. These results indicate that high population density decreased the number of weakly immunoreactive larger GH cells, and also suppressed expression of the gene encoding GH II precursor, which may result in retarded somatic growth.

Gonadotropin-releasing hormone analog (GnRHa), testosterone (T) or 17α, 20β-dihydroxy-4pregnen-3-one (DHP) was implanted in lacustrine sockeye salmon (Oncorhynchus nerka) to examine their effects on homing behavior prior to spawning. Maturing adult fish in Lake Shikotsu were captured by a set net adjacent to their natal hatchery in September and October, 1997. They were tagged, implanted with hormones and released at the center of lake. More than 70% of released fish returned to the hatchery. In September, GnRHa-implanted fish returned significantly earlier than the controls regardless of sexes. DHP also shortened homing duration in the females but not in the males. T tended to reduce homing duration in the males, however it did not have any significant effect in the females. In October, fish in all groups quickly returned within a few days after the release. Hence, the shortening effect of GnRHa on homing duration was not seen. The October fish may be already well primed by endogenous hormones. The present study showed that, in September, GnRHa consistently shortened homing duration, whereas sex steroid actions varied depending on sex in lacustrine sockeye salmon.

Adult sockeye salmon (Oncorhynchus nerka) in Lake Shikotsu were captured in September, October and November adjacent to their natal hatchery prior to spawning. They were sampled for hormones, tagged and released in the center of lake. Fish were again sampled at recapture to characterize changes in steroid hormone levels in individual migrants as well as homing percentage and duration in each month. All males returned faster than females early in the breeding season, although a half of the tagged males did not return to the natal site late in the season (November). A high percentage of females always returned, and homing duration shortened late in the season. In males, the shortening of homing duration coincided with an increase in serum testosterone (T) and 11-ketotestosterone levels. In females, the shortening of homing duration corresponded to an elevation of serum T and 17α,20β-dihydroxy-4-pregnen-3-one (DHP) levels, and a drop in serum estradiol-17β levels. Sustained administration of gonadotropin-releasing hormone analog (GnRHa, via implants) in September greatly shortened homing duration, especially in females. GnRHa treatment caused a dramatic increase in serum DHP levels in both sexes on average. Individual GnRHa-treated males which rapidly returned, however, showed higher serum T levels and lower serum DHP levels than slower returning males. The present study indicates sexual differences in homing profiles as well as shortening of homing duration following GnRHa implantation in lacustrine sockeye salmon in Lake Shikotsu which may be reflective of changes in serum steroid hormone levels.

Mature male Pacific salmon (Genus Oncorhynchus) develop a dorsal hump, as a secondary male sexual characteristic, during the spawning period. Previous gross anatomical studies have indicated that the dorsal humps of salmon are mainly composed of cartilaginous tissue (Davidson [1935] J Morphol 57:169-183.) However, the histological and biochemical characteristics of such humps are poorly understood. In this study, the detailed microstructures and components of the dorsal humps of pink salmon were analyzed using histochemical techniques and electrophoresis. In mature males, free interneural spines and neural spines were located in a line near to the median septum of the dorsal hump. No cartilaginous tissue was detected within the dorsal hump. Fibrous and mucous connective tissues were mainly found in three regions of the dorsal hump: i) the median septum, ii) the distal region, and iii) the crescent-shaped region. Both the median septum and distal region consisted of connective tissue with a high water content, which contained elastic fibers and hyaluronic acid. It was also demonstrated that the lipid content of the dorsal hump connective tissue was markedly decreased in the mature males compared with the immature and maturing males. Although, the crescent-shaped region of the hump consisted of connective tissue, it did not contain elastic fibers, hyaluronic acid, or lipids. In an ultrastructural examination, it was found that all of the connective tissues in the dorsal hump were composed of collagen fibers. Gel electrophoresis of collagen extracts from these tissues found that the collagen in the dorsal hump is composed of Type I collagen, as is the case in salmon skin. These results indicate that in male pink salmon the dorsal hump is formed as a result of an increase in the amount of connective tissue, rather than cartilage, and the growth of free interneural spines and neural spines.

Gonadotropin-releasing hormone (GnRH) is a possible secretagogue of growth hormone (GH) and somatolactin (SL) in teleosts. Effects of GnRH on the levels of pituitary mRNAs encoding GH, prolactin (PRL), and SL were therefore examined in prespawning sockeye salmon (Oncorhynchus nerka). A capsule of GnRH analog (GnRHa) was implanted into the dorsal muscle of maturing sockeye salmon for 3 weeks. The levels of hormonal mRNAs were then determined by a quantitative dot blot analysis using single-stranded sense DNA of the same sequence of mRNA as the standard. Further, we analyzed effects of GnRHa on expression of the genes encoding pituitary-specific transcription factor (Pit-1/GHF-1). Relative levels of Pit-1/GHF-1 mRNAs were estimated by Northern blot analysis, which showed specific 2- and 3-kb bands of mRNAs. GnRHa significantly increased the level of SL mRNA in the males, but not in the females, compared to the control fish. It did not induce significant increases in the levels of GH and PRL mRNAs in both the males and the females. The levels of Pit-1/GHF-1 mRNAs in the control males tended to be higher than those in the initial controls, so that GnRHa might not be effective in enhancing expression of Pit-1/GHF-1 gene, except for the level of 3-kb Pit-1/GHF-1 mRNA in the females treated with 150 microg GnRHa. The pattern of changes in the levels of Pit-1/GHF-1 mRNAs were similar to those of GH and PRL mRNAs in both the males and the females and to that of SL mRNA in the females. These results indicate that, in prespawning sockeye salmon, GnRH can stimulate SL gene expression, but probably not through the Pit-1/GHF-1-dependent system.