Individual variations in behavior and free cortisol responses to acute stress in tiger pufferfish Takifugu rubripes

Abstract:   The aim of the present study was for individual variation in behavior to be characterized and related to differences in physiological stress responses in juvenile tiger pufferfish (fugu) Takifugu rubripes . A set of siblings ( n  = 330) from wild parents was subjected to behavior tests, and some were classified into active and inactive groups ( n  = 42 each group). The active animals consisted of those individuals that kept active swimming during 6 min after transfer from one tank to another, and the inactive animals were those that stopped swimming within 6 min. Time-course acute stress responses, which were elicited by anesthesia and subsequent removal of a pectoral fin, were compared between the active and inactive fish. The stress indicators used were free and total plasma cortisol, and plasma glucose. Although no significant difference was observed at each sampling point between groups, the inactive fish required shorter recovery time from acute stress in all three indicators than the active fish. The percentage of free to total plasma cortisol in the pre-stress condition was significantly lower in the inactive group than in the active group. Our findings indicate that fugu has behavioral variation that is associated with stress responsiveness. The inactive fish are expected to be more tolerant of stress and suitable for aquaculture.     

Expression of podoplanin in various types of feline tumor tissues

Podoplanin is expressed in various human tumors where it promotes tumor progression, epithelial-mesenchymal transition, and distant metastasis. Podoplanin is also expressed in cancer-associated fibroblasts and induces tumor malignancy. The objective of this study was to evaluate podoplanin expression in various types of feline tumor tissues. Immunohistochemical analysis revealed that podoplanin was expressed in cells of 13/15 (87%) squamous cell carcinomas and 5/19 (26%) fibrosarcomas. Moreover, cancer-associated fibroblasts expressed podoplanin in most tumor types, including 18/21 (86%) mammary adenocarcinoma tissues. Our findings demonstrate that various types of feline tumor tissues expressed podoplanin, indicating the importance of the comparative aspects of podoplanin expression, which may be used as a novel research model for podoplanin biology.     

Common nucleotide sequence of structural gene encoding fibroblast growth factor 4 in eight cattle derived from three breeds

Fibroblast growth factor 4 (FGF4) is considered as a crucial gene for the proper development of bovine embryos. However, the complete nucleotide sequences of the structural genes encoding FGF4 in identified breeds are still unknown. In the present study, direct sequencing of PCR products derived from genomic DNA samples obtained from three Japanese Black, two Japanese Shorthorn and three Holstein cattle, revealed that the nucleotide sequences of the structural gene encoding FGF4 matched completely among these eight cattle. On the other hand, differences in the nucleotide sequences, leading to substitutions, insertions or deletions of amino acid residues were detected when compared with the already reported sequence from unidentified breeds. We cannot rule out a possibility that the structural gene elucidated in the present study is widely distributed in cattle. To the best of our knowledge, this is the first determination of the complete nucleotide sequence of the structural gene encoding bovine FGF4 in identified breeds.     

Changes in plasma somatolactin levels during spawning migration of chum salmon (Oncorhynchus keta)

Plasma somatolactin (SL) concentrations were examined in chum salmon in relation to gonadal maturation; immature salmon in the Bering Sea at various stages of maturation, and mature salmon during upstream migration caught at the ocean, bay and river. Plasma SL concentrations as well as plasma prolactin (PRL) and growth hormone (GH) levels in the immature fish caught in the Bering Sea were maintained essentially at similar levels. Plasma SL in mature salmon increased significantly from the fish in the ocean to the fish in the river in both sexes. Although all the fish had fully developed gonads, females completed ovulation while still in the bay, whereas final spermeation in males was achieved after entry into the river. Thus, no clear correlation was seen between plasma SL levels and final gonadal maturation. On the other hand, plasma PRL concentrations in both male and female fish were higher in the fish in the river than those in the ocean and bay, and plasma GH levels were higher in both sexes in the fish in the bay and river than those in the ocean. Plasma levels of triglycerides, glucose, free fatty acids and ionized sodium and calcium were also examined. Significant-negative correlations were seen between plasma SL and plasma ionized calcium in mature male salmon, and between plasma SL and plasma triglycerides in mature female salmon. Although our findings do not rule out the possibility of the involvement of SL in final maturation, the results indicate that SL seems to be involved at least in energy and/or calcium metabolism during the spawning migration.     

Long-lived volcanism on the lunar farside revealed by SELENE Terrain Camera

We determined model ages of mare deposits on the farside of the Moon on the basis of the crater frequency distributions in 10-meter-resolution images obtained by the Terrain Camera on SELENE (Selenological and Engineering Explorer) (Kaguya). Most mare volcanism that formed mare deposits on the lunar farside ceased at approximately 3.0 billion years ago, suggesting that mare volcanism on the Moon was markedly reduced globally during this period. However, several mare deposits at various locations on the lunar farside also show a much younger age, clustering at approximately 2.5 billion years ago. These young ages indicate that mare volcanism on the lunar farside lasted longer than was previously considered and may have occurred episodically.     

Hybrids as potential mediators spreading non-native genes: Comparison of survival,growth, and movement among native,introduced and their hybrid salmonids

Comparing multiple fitness components and potential movement of wild hybrids with their parental species is necessary to fully understand the consequences of human-mediated introgression, but studies tracking both parental species and their hybrids at the individual-level are limited. Here, we compared growth, survival and movement of sympatric introduced brook trout (BT: Salvelinus fontinalis) and native white-spotted charr (WSC: S. leucomaenis) with their hybrids (HYB) in a northern Japanese stream, using mark-recapture data (1,087 marked individuals) collected over 4 years (2013–2016). The mark-recapture data with a single cohort showed that HYB had a comparable or even higher growth rate to BT and WSC. In addition, there is no evidence that hybrid survival was lower than both parental species throughout the entire study period. Furthermore, HYB showed high mobility equivalent to WSC, while BT showed the lowest mobility. Although our previous studies have documented the reduction of BT distribution and lowered reproductive success of HYB, non-native genes can pose a threat to native WSC via relatively high survival, growth and/or mobility of HYB.     

Age‐related changes in the bovine corpus luteum function and progesterone secretion

Decreased fertility associated with maternal ageing is a well‐known critical problem, and progesterone (P4) concentration decreases during the menopause transition in women. The corpus luteum (CL) secretes P4, thereby supporting the implantation and maintenance of pregnancy. It is proposed that a bovine model is suitable for studying age‐associated decline of fertility in women because the physiology of cows is similar to that of women and cows have a greater longevity compared with other animal models. Thus, we investigated the age‐dependent qualitative changes and inflammatory responses in the bovine CL. In vivo experiment: Cows were divided into three groups, namely, young (mean age: 34.8 months), middle (80.1 months) and aged (188.9 months). Blood samples were collected on days 7 and 12 during the estrous cycle. In vitro experiments: Cows were divided into young (mean age: 27.6 months) and aged (183.1 months). The CL tissues of these groups were collected from a local slaughterhouse and used for tissue culture experiments. An in vivo experiment, plasma P4 concentration in aged cows was significantly lower than that in young cows, whereas no difference was found regarding the area of CL. An in vitro examination in the bovine CL tissues showed that the luteal P4 concentration, P4 secretion, and mRNA expression of StAR and 3β‐HSD were lower in aged cows compared with young cows, especially in the early luteal phase. However, no differences were detected in the mRNA expression of inflammation‐ and senescence‐related factors and inflammatory responses to lipopolysaccharides between the CL tissues from young and aged cows, indicating that an age‐dependent increase in inflammation is not involved in the luteal function. P4 production and secretion from the bovine CL diminish in old cows, especially during the early luteal phase, suggesting that senescence may affect the luteal function in cows.     

Effect of the application of polluted wheat (Triticum aestivum L. Thell.) straw during plowing on the transfer of radiocesium from the soil to komatsuna (Brassica rapa L. var. perviridis)

Radioactive substances were released into the environment after the nuclear accident at the Fukushima Daiichi Nuclear Power Station; this led to the contamination of the soil at Fukushima Prefecture. Mixing of organic matter with soil during plowing is known to influence radiocesium (¹³⁴Cs and ¹³⁷Cs) absorption by crops. However, the effect of mixing organic matter polluted by radioactive substances during plowing on radiocesium absorption by plants is not yet known. The aim of this study was to investigate the effect on the radiocesium absorption by komatsuna (Brassica rapa L. var. perviridis) cultivated in a 45-L container containing Andosol (14,300 Bq kg^?1) or Gray Lowland soil (33,500 Bq kg^?1) mixed with polluted wheat (Triticum aestivum L. Thell.) straw (2080 Bq kg^?1). The radiocesium concentration of the plants and the soil and the amount of exchangeable radiocesium in the soil were determined using a germanium semiconductor. The transfer of radiocesium from the soil to plants decreased by 53 and 27% in Andosol and Gray Lowland soil, respectively, after the application of 10 t ha^?1 polluted wheat straw. This reduction in the level of radiocesium transfer might be attributed to potassium contained in the wheat straw, which might compete with cesium during membrane transport and thereby block the transport of cesium from the soil solution to the roots and from the roots to the shoots. Alternatively, the applied wheat straw probably absorbed radiocesium and decreased the amount of exchangeable radiocesium in the soil. Our findings suggest that the mixing of polluted wheat straw with contaminated soil might influence the absorption of radiocesium content by agricultural products. Further studies are warranted to determine the long-term effects of the application of polluted wheat straw on the rate of radiocesium transfer to crops.     

CTLA-4 recombinant protein genetically fused to canine Fcepsilon receptor Ialpha enhances allergen specific lymphocyte responses in experimentally sensitized dogs

Vaccination with a recombinant antigen fused to a targeting molecule is a potential strategy for inducing efficient immune responses. For the therapeutic purpose of allergic diseases in dogs, a DNA construct which expresses recombinant fusion protein with two functional domains, cytotoxic T lymphocyte antigen (CTLA-4) and Fcepsilon receptor Ialpha, was developed to bridge antigen-presenting cells and IgE-allergen complex. The recombinant fusion protein expressed by the DNA construct was demonstrated to retain the ability to bind monocytes in PBMC and dog IgE, respectively. Additionally, the recombinant protein induced enhancement of allergen-induced lymphoproliferation in experimentally sensitized dogs under conditions of suboptimal allergen stimulation. These results indicated that the DNA construct could enhance allergen-induced immune responses in vivo, implying its usefulness for perspective application in immunotherapy in dogs.     

Cytokine profiles of peripheral blood mononuclear cells from dogs experimentally sensitized to Japanese cedar pollen

Japanese cedar (Cryptomeria japonica, CJ) pollinosis is mediated by type-I hypersensitivity and induces seasonal rhinitis and conjunctivitis in humans. Previous studies showed that dogs could be experimentally sensitized with CJ pollen. In this study, we carried out quantitative analysis of mRNA levels of various cytokines in the peripheral blood mononuclear cells (PBMC) of 12 dogs experimentally sensitized to Japanese cedar pollen. Experimental sensitization was carried out by injection of crude CJ pollen extract with aluminium hydroxide gel. The expression levels of interleukin (IL)-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, IL-18, interferon (IFN)-gamma, transforming growth factor (TGF)-beta(1), and tumor necrosis factor (TNF)-alpha mRNAs in the PBMC were quantified using a real-time sequence detection system. In the PBMC tested without culture, the expression levels of IL-8 and TNF-alpha mRNAs in experimentally sensitized dogs were significantly higher than those in control dogs. The expression level of IFN-gamma mRNA in the sensitized group was significantly lower than that in the control group. When the PBMCs were cultured in the presence of CJ pollen extract, the level of IL-4 mRNA expression was markedly increased in the PBMC from the experimentally sensitized dogs. In the PBMC stimulated with the CJ pollen extract, the expression level of IL-2 mRNA in the sensitized group was also significantly higher than that in the control group. Our data indicated that a Th2 response and proliferation of PBMC occur in response to the sensitizing antigen in dogs experimentally sensitized with CJ pollen, and revealed the presence of antigen-specific Th2 cells in this canine model. In addition, the expression levels of the mRNAs encoding proinflammatory cytokines were shown to be elevated after CJ pollen sensitization, indicating the activation of monocytes and macrophages.