Interferon-gamma is causatively involved in experimental inflammatory bowel disease in mice.

Cytokines may be crucially involved in the pathogenesis of inflammatory bowel diseases (IBD), but it remains controversial whether interferon (IFN)-gamma, a typical proinflammatory cytokine, is an essential mediator to cause the disorders. In the present study, IFN-gamma(-/-) and wild-type (WT) C57BL/6 mice were fed 2.5% dextran sodium sulphate (DSS) in drinking water for 7 days, in order to investigate DSS-induced intestinal inflammation. The DSS-treated WT mice exhibited a robust production of IFN-gamma in the gut, a remarkable loss of body weight, as well as high rate of mortality (60%). In striking contrast, IFN-gamma deficient mice did not develop DSS-induced colitis, as indicated by the maintenance of body weight and survival rate of 100%. Severe intestinal inflammation was demonstrated exclusively in WT animals in terms of the shortening of the bowel as well as the elevation of the disease activity index, myeloperoxidase (MPO) activity and serum haptoglobin level. Histological study of DSS-treated WT intestine revealed disruption of mucosal epithelium and massive infiltration of inflammatory cells, while the organ from IFN-gamma(-/-) mice remained virtually normal in appearance. Enzyme-linked immunosorbent assay (ELISA) analyses indicated abundant production of three chemokines, i.e. monokine induced by interferon-gamma (MIG), interferon-inducible protein 10 (IP-10) and monocyte chemoattractant protein-1 (MCP-1), in the DSS-irritated intestine of WT but not of IFN-gamma(-/-) mice. The present results demonstrate clearly that IFN-gamma plays indispensable roles in the initiation of DSS colitis, and some chemokines are produced in an IFN-gamma-dependent fashion.

Molecular identification and immunohistochemical localization of atrial natriuretic peptide in the heart of the dromedary camel (Camelus dromedarius).

Atrial and B-type natriuretic peptide (ANP and BNP) are cardiac hormones synthesized and secreted by the myoendocrine cells of the heart. They exert potent actions on body fluid balance. Since various body organs including the heart are under high physiological stress during water and food deprivation in the desert nomads, we intended to perform molecular biological and histological studies of ANP in the heart of the dromedary camel Camelus dromedarius. Initially, we isolated cDNAs encoding ANP from the atrium and BNP from the atrium and ventricle of the dromedary camel. Putative mature ANP, deduced from the cDNA sequence, was identical to that of human and pig ANP, but the putative mature BNP was more diverse and was most similar to pig BNP (94% identity). Thus, we used antisera raised against human ANP that did not cross-react with pig BNP in the subsequent immunohistochemical studies. The ANP-expressing myoendocrine cells are most concentrated in the right atrium, to a lesser extent in the left atrium, and almost absent in the left ventricle. The immuno-positive cells are scattered uniformly in each region and are characterized by the presence of immunoreactive granular deposits around the nucleus. The left atrium comprises some ramifications of conductive cells (Purkinje fibers), some of which also contained ANP-immunoreactive granules. At the electron microscopic level, myoendocrine cells possessed secretory granules primarily in the perinuclear zone and a well-developed Golgi apparatus. The present study is the first comprehensive report dealing with the molecular cloning and immunohistochemical localization of ANP in the heart of a desert dwelling mammal.

Functional organization of preoptic vasotocin and isotocin neurons in the brain of rainbow trout: central and neurohypophysial projections of single neurons.

Preoptic magnocellular neurosecretory cells (NSCs) in the brain of rainbow trout show synchronization of periodic Ca(2+) pulses, patterns of which differ between vasotocin (VT) and isotocin (IT) neurons. To provide neuroanatomical bases of the synchronized periodic Ca(2+) pulses and their biological implications, we examined the organization of preoptic VT and IT neurons in the brain of rainbow trout. The cytoarchitecture of the preoptic neurosecretory system was characterized by a confocal double-color immunofluorescence. Two to five VT neurons, and also IT neurons, aggregate to form cell-type specific clusters. VT clusters tend to localize medially, while IT clusters laterally. VT neurons are closely apposed at the proximal neuronal processes. A Golgi-like immunohistochemistry demonstrated that VT and IT fibers distribute widely in the brain, such as ventral telencephalon, diencephalon, and various mesencephalic structures, in addition to the neurohypophysial projections. Projections from single VT and IT neurons were examined by an intracellular staining with biocytin injection in a sagittally hemisected brain preparation, which contains the entire forebrain region. Single VT and IT neurons project toward the pituitary and the extrahypothalamic regions. Some IT neurons, but not VT neurons, were dye-coupled. These results support the idea that the same types of NSCs are connected to form cell-type-specific networks responsible for the synchronization of periodic Ca(2+) pulses. The organization of the preoptic neurosecretory system shown in the present study is suitable for the simultaneous control of neurohypophysial and extrahypothalamic outputs through the synchronization of electrical activity.

Synchronized periodic Ca2+ pulses define neurosecretory activities in magnocellular vasotocin and isotocin neurons.

The electrical activity of magnocellular neurosecretory cells (NSCs) is correlated with the release rates of neurohypophysial hormones. NSCs may control their secretory activity in a cooperative manner by changing their electrical activity in response to changes in the internal milieu. In the present study, we applied confocal Ca(2+) imaging to a sagittally hemisected rainbow trout brain to simultaneously monitor the neuronal activity of a number of NSCs. We found that NSCs in vitro showed synchronized pulsatile elevations of intracellular Ca(2+) levels at regular intervals. Double immunostaining of vasotocin (VT) and isotocin (IT) after the confocal imaging clarified that each of the VT and IT neuronal populations showed a distinct pattern of periodic Ca(2+) pulses. Simultaneous cell-attached patch recordings ensured that individual Ca(2+) pulses were associated with a phasic burst firing. Depolarizing stimuli by increasing the extracellular K(+) concentration from 5 to 7-9 mm reversibly shortened the interpulse intervals in both VT and IT neurons. Interpulse intervals but not durations of pulses were shortened by hypo-osmotic stimuli and prolonged by hyperosmotic stimuli, consistent with the osmoregulatory function of teleost NSCs. We therefore hypothesize that NSCs use intervals of synchronized periodic burst discharges to fit the levels of secretory activity to physiological requirements.

Prepubertal increases in the levels of two salmon gonadotropin-releasing hormone mRNAs in the ventral telencephalon and preoptic area of masu salmon.

Ontogenic changes in the expression levels of two salmon gonadotropin-releasing hormone genes (sGnRH-I and -II) were examined in the forebrain region including the ventral telencephalon and preoptic area of masu salmon by competitive reverse transcription-polymerase chain reaction (RT-PCR). Two genes showed similar expression patterns throughout the lifetime in both sexes, although the levels of sGnRH-II mRNA were about 20 times higher than those of sGnRH-I mRNA. In males, the levels of sGnRH mRNAs increased at the beginning of the second year and reached their maximum in the autumn. The levels decreased gradually until the autumn of the third year when fish sexually matured. In females, the levels reached their maximum in the first autumn and fluctuated considerably along with the seasons in the third year. These results suggest that, in the salmon brain, sGnRH genes are activated long before the sexual maturation under sexually different control mechanisms.

Expression of Fushi tarazu factor 1 homolog and Pit-1 genes in the pituitaries of pre-spawning chum and sockeye salmon.

Fushi tarazu factor-1 (FTZ-F1) and Pit-1 are major pituitary transcription factors, controlling expression of genes coding for gonadotropin (GTH) subunits and growth hormone/prolactin/somatolactin family hormone, respectively. As a first step to investigate physiological factors regulating gene expression of these transcription factors, we determined their mRNA levels in the pituitaries of chum salmon (Oncorhynchus keta) at different stages of sexual maturation. FTZ-F1 gene expression was increased in males at the stage before spermiation, where the levels of GTH alpha and IIbeta subunit mRNAs were elevated. Pit-1 mRNA showed maximum levels at the final stage of sexual maturation in both sexes, when expression of somatolactin gene peaked. To clarify whether gonadotropin-releasing hormone (GnRH) is involved in these increases in FTZ-F1 and Pit-1 gene expression, we examined effects of GnRH analog (GnRHa) administration on their gene expression in maturing sockeye salmon (Oncorhynchus nerka). GnRHa stimulated Pit-1 gene expression in females only, but failed to stimulate FTZ-F1 gene expression in both sexes. The up-regulated expression of FTZ-F1 and Pit-1 genes at the pre-spawning stages suggest that the two transcription factors have roles in sexual maturation of salmonids. Physiological factors regulating gene expression of FTZ-F1 and Pit-1 are discussed in this review.

Signal transduction pathways and transcription factors involved in the gonadotropin-releasing hormone-stimulated gonadotropin subunit gene expression.

Gonadotropin-releasing hormone (GnRH) stimulates gonadotropin (GTH) subunit gene expression via G protein-coupled membrane receptors. GnRH-stimulated GTH subunit gene expression is mediated by protein kinase C (PKC) and Ca(2+) signaling pathways. Recent numerous reports on signal transduction pathways which are involved in GnRH stimulation of mammalian GTH subunit genes showed differential sensitivity of GTH subunit genes to the two signaling pathways. Our recent studies on salmon GTH (sGTH) IIbeta subunit gene showed that its stimulation by GnRH is dependent on the PKC pathway. Furthermore, gel retardation and mutagenesis studies suggested that pituitary homeo box 1 (Ptx1) and Sp1 mediate the GnRH-induced PKC signaling on the sGTHIIbeta gene. However, both PKC and Ca(2+) pathways are involved in the GnRH-stimulated GTH alpha and LHbeta genes. Different preference to the pathways were often reported in a certain GTH subunit gene in different circumstances, suggesting that molecular targets of the two signaling pathways are different. Ets-related factor and cAMP response element binding protein have been proposed as targets of GnRH signaling on GTH alpha genes. Sp1 and early growth response protein 1 play pivotal roles in GnRH-stimulated LHbeta gene expression in synergism with steroidogenic factor-1 and Ptx1. Activating protein-1 mediates GnRH-induced PKC signaling to stimulate FSHbeta gene expression. Therefore, divergent transcription factors are involved in GnRH stimulation of GTH subunit gene expression, and molecular mechanisms of GnRH stimulation may be partially conserved between sGTH IIbeta and mammalian LHbeta genes.

Effects of gonadotropin-releasing hormone analog on expression of genes encoding the growth hormone/prolactin/somatolactin family and a pituitary-specific transcription factor in the pituitaries of prespawning sockeye salmon.

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.

Seasonal changes in expression of neurohypophysial hormone genes in the preoptic nucleus of immature female masu salmon.

In relevance to osmoregulatory and reproductive functions, activity of the hypothalamic magnocellular neurosecretory system may vary seasonally in teleosts. The changes in the expression of vasotocin (VT) and isotocin (IT) genes were thus studied by an in situ hybridization technique and an immunohistochemical avidin-biotin complex method in immature female masu salmon (Oncorhynchus masou). The plasma levels of testosterone and estradiol were also measured by enzyme immunoassay. Fish were sampled in March, May, August, and November 1994 and January 1995. The intensity of autoradiographic hybridization signals and immunoreactivity were determined in individual neurosecretory cells (NSC) in the rostroventral, middle, and dorsocaudal regions of the magnocellular part of the preoptic nucleus (PM). The VT hybridization signals and immunoreactivity were high in November, along with the elevation of plasma levels of testosterone and estradiol. These results suggest that sex steroid hormones are involved in seasonal regulation of VT gene expression. The hybridization signals for IT mRNA were increased in May and decreased in November, whereas IT immunoreactivity was low in March and high in November. NSCs thus showed seasonal variations in the intensity of hybridization signals for VT and IT mRNAs and immunoreactivity of VT and IT, although the patterns of changes were different between VT and IT. VT and IT genes may be seasonally expressed under different regulatory mechanisms.

Differences in seasonal expression of neurohypophysial hormone genes in ordinary and precocious male masu salmon.

Our previous study showed the seasonal variations in expression of vasotocin (VT) and isotocin (IT) genes in preoptic magnocellular neurons of female masu salmon (Oncorhynchus masou). The changes in the level of VT mRNA were coincident with those in plasma testosterone and estradiol levels. In the present study, generality of this phenomenon in salmonid was verified in males. We examined changes in expression of VT and IT genes by an in situ hybridization technique and an immunohistochemical avidin-biotin complex method in the preoptic nuclei of ordinary and precocious male masu salmon. Plasma levels of testosterone and estradiol were measured by enzyme immunoassay. Fish were sampled in March, May, August, and November 1994 and January 1995. The intensities of hybridization signals for VT and IT mRNAs, as well as immunoreactivity of VT and IT, showed seasonal variations, although the profiles were different between the ordinary and precocious males. In the ordinary males, the intensities of hybridization signals for VT and IT mRNAs were high in January. These strong hybridization signals, representing elevation of VT and IT gene expression, were accompanied by increases in plasma levels of testosterone and estradiol. However, in precocious males, changes in VT and IT mRNA levels were not coincident with variation of plasma levels of sex steroid hormones. The sensitivity to sex steroid hormones of VT and IT gene expression may be different between the ordinary and precocious male masu salmon.

Changes in the levels of mRNAs for GH/prolactin/somatolactin family and Pit-1/GHF-1 in the pituitaries of pre-spawning chum salmon.

Changes in the levels of pituitary mRNAs encoding GH, prolactin (PRL) and somatolactin (SL) were determined in pre-spawning chum salmon (Oncorhynchus keta) caught at a few key points along their homing pathway in 1994 and 1995. Furthermore, we analyzed relationships between expression of pituitary-specific POU homeodomain transcription factor (Pit-1/GHF-1) and GH/PRL/SL family genes. In 1994, seawater (SW) fish and matured fresh-water (FW) fish were sequentially captured at two points along their homing pathway, the coast and the hatchery. In addition to these two points, maturing FW fish were captured at the intermediate of the two points in 1995. The levels of hormonal mRNAs were determined by a quantitative dot blot analysis using single-stranded sense DNA as the standard. Relative levels of Pit-1/GHF-1 mRNAs were estimated by Northern blot analysis. In 1994, the levels of GH/PRL/SL family mRNAs except for PRL mRNA in the male FW fish were 1.8-4 times higher than those in the SW fish. In 1995, the level of PRL mRNA was somewhat sharply elevated in the maturing FW fish soon after entry into the FW environment, while that of SL mRNA was gradually increased during upstream migration from the coast to the hatchery. The levels of 3 kb Pit-1/GHF-1 mRNA in the FW fish were higher than those in the SW fish in both 1994 and 1995. The present results indicate that expression of genes for the GH/PRL/SL family and Pit-1/GHF-1 is coincidentally enhanced in homing chum salmon. Moreover, the present study suggests that expression of the SL gene is elevated with sexual maturation, whereas that of PRL gene is elevated with osmotic change during the final stages of spawning migration.

Expression of salmon corticotropin-releasing hormone precursor gene in the preoptic nucleus in stressed rainbow trout.

The behavior of genes encoding the corticotropin-releasing hormone (CRH) precursor in response to stress has not been extensively studied in teleosts. To clarify this problem, CRH cDNAs were isolated from a hypothalamic cDNA library of sockeye salmon, Oncorhynchus nerka, by screening with PCR products amplified from the hypothalamic mRNA with primers deduced from the sequence of the sucker CRH precursor. Two types of PCR products with a high degree of sequence homology were identified (CRH-I and CRH-II). A cDNA encompassing the entire coding sequence of the salmon CRH-I precursor was isolated. The salmon CRH-I cDNA encodes a 167-amino-acid precursor, which consists of a signal sequence, a cryptic peptide, and the carboxyl terminal 41-amino-acid sequence of CRH. The deduced amino acid sequence of salmon CRH peptide exhibits 66 to 80% homology with mammalian, Xenopus, and sucker CRHs, whereas it shows about 50% homology with sucker, carp, or sole urotensin I, a CRH-related neuropeptide in teleost fish. In situ hybridization histochemistry demonstrated CRH mRNA-positive perikarya in the preoptic nucleus in rainbow trout, Oncorhynchus mykiss, when the fish were stressed by confinement. Adjacent sections hybridized with probes for salmon vasotocin (VT) precursor showed many VT mRNA-positive neurons also in the preoptic nucleus, suggesting a colocalization of CRH and VT mRNAs in the same magnocellular neurons in the rainbow trout brain. The present results suggest that CRH may have important roles in the control of stress responses in salmonid fish.

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.

Changes in expression of neurohypophysial hormone genes during spawning migration in chum salmon, Oncorhynchus keta.

We analyzed changes in the hypothalamic levels of vasotocin (VT) and isotocin (IT) mRNA in chum salmon during spawning migration to the Ishikari river. The fish were caught at Atsuta, a fisherman's village facing the Ishikari bay, and at Chitose, an upstream branch of the Ishikari river. The former are referred to as sea water (SW) fish, and the latter as freshwater (FW) fish. The levels of VT and IT mRNA in the forebrains were determined by quantitative Northern blot analysis using single-stranded DNA with the same mRNA sequences as the standards. Levels of VT mRNA were higher in the FW males than the FW females, although no such difference was seen in the SW fish. Changes in the levels of VT mRNA were markedly different in males and females. In the males, no significant differences were seen in the levels of VT-I and VT-II mRNA between the SW and FW fish. However, in the females, the levels of VT mRNA in the FW fish were significantly lower than those in the SW fish. Changes in the levels of IT-I and IT-II mRNA were essentially similar in the males and females. These results suggest that the control of VT gene expression is different in males and females during spawning migration, although the neuroendocrine mechanism is not known.

cDNA for ribosomal protein S2 in sockeye salmon, Oncorhynchus nerka.

We isolated a cDNA encoding ribosomal protein S2 in sockeye salmon, Oncorhynchus nerka, using a reverse transcriptase-polymerase chain reaction (RT-PCR) method. The cDNA encoding ribosomal protein S2 is composed of 933 nucleotides, and has a 5'-noncoding sequence of 9 bases, a 885 base open reading frame coding for a 294 amino acid polypeptide, and a 39 base 3'noncoding sequence. The amino acid sequence of sockeye salmon S2 protein deduced from the nucleotide sequence is highly homologous to those from the rat (86.1%) and Drosophila melanogaster (73.6%). The N-terminal region of S2 protein is rich in arginine-glycine sites, including eight tandem repeats, and has two consecutive copies of the RGGF motif. The sequences are considered to be requisites for nucleolar localization and binding to RNA for nucleolar proteins. Southern blot analysis indicates that there may be only a single copy of the S2 gene, which is a multiple copy gene in the rat and the fruit fly. Northern blot analysis shows that the S2 gene is expressed in the brain, pituitary, heart, liver kidney, muscle, testis and ovary of sockeye salmon.

Hydropathy profiles of predicted thyrotropin-releasing hormone precursors are highly conserved despite low similarity of primary structures.

Two types of cDNAs encoding thyrotropin-releasing hormone (TRH) precursors (TRH-A and TRH-B) were amplified from hypothalamic mRNA of sockeye salmon by reverse transcriptase-polymerase chain reaction (RT-PCR). The amplification was achieved using two primers which correspond to TRH progenitor sequence (Lys/Arg-Arg-Gln-His-Pro-Gly-Lys/Arg-Arg). A full length cDNA encoding TRH-A was obtained by 5'- and 3'-RACE methods. It has a length of 1324 base pairs (bp) that contains sequences of 5' and 3' untranslated regions and an open reading frame of 259 codons. The sockeye salmon TRH-A deduced from the nucleotide sequence tandemly contains 8 copies of TRH progenitor sequences. Another cDNA which encodes a part of TRH-B consists of 242 bp, and the sequence homology between TRH-A and -B cDNAs is 90%. The result of Southern blot analysis of sockeye and masu salmon genomic DNAs supported the evidence that there are at least two TRH genes in the salmonid. A RT-PCR analysis of TRH gene expression in various tissues of sockeye salmon showed that strong expression was observed only in the brain. The primary structure of the sockeye salmon TRH-A shares low similarity to those of human, rat and Xenopus TRH precursors (35, 27 and 44%, respectively). However, their hydropathy profiles were almost the same with each other. The profile of sockeye salmon TRH-A showed the presence of two discrete hydrophobic regions, one in the N-terminal region which corresponds to the signal peptide and the other in the C-terminal region. All of the repetitive TRH progenitor sequences are included in three hydrophilic regions easily recognizable. The present results thus suggest that the three-dimensional structures of TRH precursors are highly conserved, although the primary structures of TRH precursors have diverged through the evolutionary pathway of vertebrates.

Two types of cDNAs encoding proopiomelanocortin of sockeye salmon, Oncorhynchus nerka.

To investigate regulatory mechanisms of proopiomelanocortin (POMC) gene expression in sockeye salmon, we have isolated and characterized cDNAs encoding two types of sockeye salmon POMC, which are referred to as ssPOMC-A and -B. Two types of PCR products were amplified from total RNA of sockeye salmon pituitaries by use of rainbow trout sequences. Full length cDNA clones encoding ssPOMC-A and ssPOMC-B were obtained from a pituitary cDNA library of sockeye salmon using the PCR products as probes. The ssPOMC-A and -B cDNAs have a length of 1072 and 1709 bps, respectively. Northern blot analysis showed that both ssPOMC-A and -B mRNAs were expressed only in the pituitary, and their sizes were about 1.2 kb and 1.8 kb, respectively. The presence of two ssPOMC genes was confirmed by Southern blot analysis of genomic DNA obtained from a single sockeye salmon. The deduced amino acid sequences of the ssPOMC-A and -B contained 230 and 226 residues, respectively. The amino terminal of beta-endorphin in ssPOMC-B which corresponds to Met-enkephalin domain is YSGFM, which is different from YGGFM of Met-enkephalin found in many other vertebrate species. The homology of nucleotide sequences between ssPOMC-A and -B is 59% in the entire coding region, whereas alpha-MSH coding regions are highly homologous (91%). Although the deduced amino acid sequences of ssPOMs show 43% overall similarity, their hydropathy profiles are coincident with those of several other vertebrate species, particularly the amino terminal of N-terminal peptide (NPP) shows almost the same pattern with other vertebrate NPPs.

Cytophysiology of gonadotropin-releasing-hormone neurons in chum salmon (Oncorhynchus keta) forebrain before and after upstream migration.

Cytophysiology of gonadotropin-releasing-hormone neurons in chum salmon (Oncorhynchus keta) was examined before and after upstream migration by an immunocytochemical technique with a specific antiserum to salmon gonadotropin-releasing hormone and an in situ hybridization technique with an oligonucleotide encoding salmon gonadotropin-releasing hormone precursor (pro-salmon gonadotropin- releasing hormone). In the forebrain (olfactory nerve, olfactory bulb, telencephalon, and preoptic area), salmon gonadotropin-releasing hormone-immunoreactive neurons and neurons showing signals for pro-salmon gonadotropin-releasing hormone mRNA were compared between fish from the coastal sea and those from the spawning ground. Neurons in the dorsal region of the olfactory nerve and in the ventral region of the transitional area between olfactory nerve and olfactory bulb showed strong salmon gonadotropin-releasing-hormone immunoreactivity and strong hybridization signals in fish from the coastal sea, but these activities and signals were not observed or were decreased in number in fish from the spawning ground. The neurons in the olfactory bulb, telencephalon, and preoptic area consistently revealed salmon gonadotropin-releasing-hormone immunoreactivity and hybridization signals, and the hybridization signals of salmon gonadotropin-releasing hormone in the telencephalon and the preoptic area were stronger in fish from the spawning ground than in those from the coastal sea. These findings suggest that salmon gonadotropin-releasing-hormone neurons in the olfactory nerve and the transitional area between olfactory nerve and olfactory bulb have different patterns of hormone production than those in the telencephalon and the preoptic area.