α1 and ß3 adrenergic receptor-mediated excitatory effects of adrenaline on the caudal neurosecretory system (CNSS) in olive flounder, Paralichthys olivaceus.

Adrenaline is one of the most important neurotransmitters in the central nervous system and is produced during stress. In this study, we investigated the modulatory role of adrenaline and adrenergic receptors on the neuroendocrine Dahlgren cells in the caudal neurosecretory system (CNSS) of olive flounder. Ex vivo electrophysiological recordings revealed that adrenaline significantly increased the firing frequency and altered the firing pattern of Dahlgren cells. Moreover, treatment with adrenaline led to a significant upregulation of ion channels and major hormone secretion genes in CNSS at the mRNA levels. Additionally, treatment with adrenaline resulted in a significantly elevation in the expression levels of α1- and ß3-adrenergic receptors. Furthermore, the ß3-adrenergic receptor antagonist exerts a significant inhibitory effect on adrenaline-induced enhancement firing activities of Dahlgren cells, whereas the α1-adrenergic receptor antagonist displays a comparatively weaker inhibitory effect. Additionally, the enhanced firing activity induced by adrenaline could be effectively suppressed by both α1- and ß3-adrenergic receptor antagonists. Taken together, these findings provide strong evidence in favor of the excitatory effects of adrenaline through α1 and ß3 adrenergic receptors in CNSS to stimulate the secretion of stress-related hormones, ß3-adrenergic receptor plays a more dominant role in the modulation of firing activities of Dahlgren cells by adrenaline and thereby regulates the stress response in olive flounder.

Switch from fight-flight to freeze-hide: The impacts of severe stress and brain serotonin on behavioral adaptations in flatfish.

Animals often experience changes in their environment that can be perceived as stressful. Previous evidence indicates that different individuals may have distinct stress responses. The role of serotonin (5-HT) in stress adaptation is well established, but its relationship with different defense strategies and the persistence of physiological and behavioral responses in different individuals during repeated acute stress remain unclear. In this study, using olive flounder (Paralichthys olivaceus) as a model, we analyzed the relationship between boldness and neurotransmitter 5-HT activity. We found that 5-HT suppression with 5-HT synthesis inhibitor p-chlorophenylalanine (pCPA) and 5-HT receptor subtype 1A (5-HT1A) antagonist WAY-100635 increased their oxygen consumption rates and the boldness of shy individuals. We determined the metabolic and behavioral changes in bold and shy individuals to repeated acute stress. The results suggest that bold individuals switch on passive "energy-saving" personality by changing their defense behavior from "fight-flight" to "freeze-hide" during a threat encounter, which manifests high behavioral plasticity. Both behavioral types decreased their spontaneous activity levels, which were also strengthened by limiting metabolic rate. Interestingly, treatment with pCPA and WAY-100635 before stress procedure attenuated stress and increased the boldness across diverse behavioral types. This study provides the initial empirical evidence of how perception of stress impacts both individual defense behavior and personality in this species. These findings can enhance our comprehension of individual variability and behavioral plasticity in animals, thereby improving our ability to develop effective adaptive management strategies.

The excitatory effect of 5-HT1A and 5-HT2B receptors on the caudal neurosecretory system Dahlgren cells in olive flounder, Paralichthys olivaceus.

The neurotransmitter 5-hydroxytryptamine (5-HT, serotonin) plays an essential role in the regulation of neural activity via multiple receptors. Here, we investigated the functional role of serotoninergic input on the Dahlgren cell population in the caudal neurosecretory system (CNSS) of olive flounder. In this study, the effect of 5-HT on the firing activity of Dahlgren cells was explored in terms of changes in firing frequency and firing pattern using multicellular recording electrophysiology ex vivo, and the role of several 5-HT receptor subtypes in the regulation was determined. The results revealed that 5-HT increased the firing frequency in a concentration-dependent manner and altered the firing pattern of Dahlgren cells. The effect of 5-HT on the firing activity of Dahlgren cells was mediated through the 5-HT1A and 5-HT2B receptors, selective agonists of both receptors effectively increased the firing frequency of Dahlgren cells, and selective receptor antagonists could also effectively inhibit the increase in firing frequency caused by 5-HT. In addition, the mRNA levels of major signaling pathway-related genes, ion channels, and major secretion hormone genes were significantly upregulated in CNSS after treatment with 5-HT. These findings demonstrate that 5-HT acts as an excitatory neuromodulator on Dahlgren cells and enhances neuroendocrine activity in CNSS.

Electronic Structure Manipulation of the Mott Insulator RuCl3 via Single-Crystal to Single-Crystal Topotactic Transformation.

The core task for Mott insulators includes how rigid distributions of electrons evolve and how these induce exotic physical phenomena. However, it is highly challenging to chemically dope Mott insulators to tune properties. Herein, we report how to tailor electronic structures of the honeycomb Mott insulator RuCl3 employing a facile and reversible single-crystal to single-crystal intercalation process. The resulting product (NH4 )0.5 RuCl3 ⋅1.5 H2 O forms a new hybrid superlattice of alternating RuCl3 monolayers with NH4 + and H2 O molecules. Its manipulated electronic structure markedly shrinks the Mott-Hubbard gap from 1.2 to 0.7 eV. Its electrical conductivity increases by more than 103 folds. This arises from concurrently enhanced carrier concentration and mobility in contrary to the general physics rule of their inverse proportionality. We show topotactic and topochemical intercalation chemistry to control Mott insulators, escalating the prospect of discovering exotic physical phenomena.

Interleukin 1 receptor type I (IL-1RI) is involved in the innate immune response of olive flounder (Paralichthys olivaceus) to resist pathogens.

The pleiotropic cytokine IL -1 is involved in important immune responses such as thymocyte proliferation and B cell growth and differentiation. Activation of the IL -1 pathway requires its functional receptor IL -1RI, making IL -1RI the critical point of the IL -1 pathway. In-depth study of IL -1RI will help to understand the immune mechanism involved in IL -1. In this study, we identified the cDNA of the IL -1RI gene of olive flounder (PoIL-1RI). The total length of the PoIL-1RI cDNA is 2490 bp, the open reading frame is 1689 bp long and encodes a protein of 562 amino acids. The protein has three Ig domains and a typical TIR domain, as in other mammals and fish. We found that PoIL-1RI is widely expressed in the tissues studied and shows a significant immune response after stimulation with bacteria and pathogen-associated molecular patterns (PAMPs) both in vitro and in vivo. After PoIL-1RI was overexpressed in olive flounder embryonic cell line (FEC), pro-inflammatory cytokines (IL -1ß, IL -6, IL -8, TNF-α) and interferon (IFN-α, IFN-γ) were significantly upregulated. And we found that after overexpressing PoIL-1RI in FEC, the antibacterial ability of FEC was significantly stronger than that of the control group, and we found that overexpression of PoIL-1RI gene significantly increased the activity of NF-κB signaling pathway. These results suggest that PoIL-1RI plays an important role in innate immune response.

GABAA receptor-mediated inhibition of Dahlgren cells electrical activity in the olive flounder, Paralichthys olivaceus.

γ-Aminobutyric acid (GABA) is a major inhibitory neurotransmitter in the central nervous system. We investigated its potential role as a neurotransmitter in the neuroendocrine Dahlgren cell population of the caudal neurosecretory system (CNSS) of the flounder Paralichthys olivaceus. The application of GABA in vitro resulted in a decrease in electrical activity of Dahlgren cells, followed by an increase of the number of silent cells, together with a decreased firing frequency of all three activity patterns (tonic, phasic, bursting). GABAA receptor agonist etomidate decreased Dahlgren cell firing activity, in a similar way to GABA. The response to GABA was blocked by the GABAA receptor antagonist bicuculline. GABAA receptor gamma2 subunit (Gabrg2) and chloride channel (Clcn2) mRNA expression were significantly upregulated in the CNSS after GABA superfusion. These data suggest that GABA may modulate CNSS activity in vivo mediated by GABAA receptors.

Neuromodulatory effects of GnRH on the caudal neurosecretory Dahlgren cells in female olive flounder.

Gonadotropin-releasing hormone (GnRH) is considered a key player in reproduction. The caudal neurosecretory system (CNSS) is a unique neurosecretory structure of fish that may be involved in osmoregulation, nutrition, reproduction, and stress-related responses. However, a direct effect of GnRH on Dahlgren cells remains underexplored. Here, we examined the electrophysiological response of Dahlgren cell population of the CNSS to GnRH analog LHRH-A2 and the transcription of related key genes of CNSS. We found that GnRH increased overall firing frequency and may be changed the firing pattern from silent to burst or phasic firing in a subpopulation of Dahlgren cells. The effect of GnRH on a subpopulation of Dahlgren cells firing activity was blocked by the GnRH receptor (GnRH-R) antagonist cetrorelix. A positive correlation was observed between the UII and GnRH-R mRNA levels in CNSS or gonadosomatic index (GSI) during the breeding season. These findings are the first demonstration of the ability of GnRH acts as a modulator within the CNSS and add to our understanding of the physiological role of the CNSS in reproduction and seasonal adaptation.

Exceptionally High Average Power Factor and Thermoelectric Figure of Merit in n-type PbSe by the Dual Incorporation of Cu and Te.

Thermoelectric materials with high average power factor and thermoelectric figure of merit (ZT) has been a sought-after goal. Here, we report new n-type thermoelectric system CuxPbSe0.99Te0.01 (x = 0.0025, 0.004, and 0.005) exhibiting record-high average ZT ∼ 1.3 over 400-773 K ever reported for n-type polycrystalline materials including the state-of-the-art PbTe. We concurrently alloy Te to the PbSe lattice and introduce excess Cu to its interstitial voids. Their resulting strong attraction facilitates charge transfer from Cu atoms to the crystal matrix significantly. It follows the increased carrier concentration without damaging its mobility and the consequently improved electrical conductivity. This interaction also increases effective mass of electron in the conduction band according to DFT calculations, thereby raising the magnitude of Seebeck coefficient without diminishing electrical conductivity. Resultantly, Cu0.005PbSe0.99Te0.01 attains an exceptionally high average power factor of ∼27 µW cm-1 K-2 from 400 to 773 K with a maximum of ∼30 µW cm-1 K-2 at 300 K, the highest among all n- and p-type PbSe-based materials. Its ∼23 µW cm-1 K-2 at 773 K is even higher than ∼21 µW cm-1 K-2 of the state-of-the-art n-type PbTe. Interstitial Cu atoms induce the formation of coherent nanostructures. They are highly mobile, displacing Pb atoms from the ideal octahedral center and severely distorting the local microstructure. This significantly depresses lattice thermal conductivity to ∼0.2 Wm-1 K-1 at 773 K below the theoretical lower bound. The multiple effects of the dual incorporation of Cu and Te synergistically boosts a ZT of Cu0.005PbSe0.99Te0.01 to ∼1.7 at 773 K.

Inhibitory role of taurine in the caudal neurosecretory Dahlgren cells of the olive flounder, Paralichthys olivaceus.

Taurine plays role in neural development and physiological functions such as endocrine regulation in the central nervous system (CNS), and it is one of the most abundant free amino acid there. We investigated its potential effect as a neurotransmitter in the group of neuroendocrine Dahlgren cells at flounder Paralichthys olivaceus caudal neurosecretory system (CNSS). The application of taurine in vitro led to a reduction in electrical activity of Dahlgren cells, followed by a rise in the number of silent cells, at the same time the frequency of all three activity patterns (tonic, phasic, bursting) in Dahlgren cells was reduced. Both strychnine (a glycine receptor antagonist) and bicuculline (a GABAA receptor antagonist) can block the response to taurine separately. Transcriptome sequencing analysis showed the existence of glycine receptor (GlyR) and GABAA receptor (GABAAR) in the flounder CNSS, and the GlyR, GABAAR, and Cl- channel mRNA expression were significantly raised after taurine superfusion according to quantitative RT-PCR results. These data indicate that taurine may mediate Dahlgren cell population of CNSS activity in vivo through GlyR and GABAAR, thereby, regulating stress-response.

Shedding Light on the Intrinsic Characteristics of 3D Distorted Fluorite-Type Zirconium Tellurite Single Crystals.

Transition-metal tellurites have motivated growing research interest in both fundamental and applied chemistry, and the corresponding single crystals could serve as rich and fascinating platforms to regulate, explore, and elucidate the intrinsic characteristics of different structures from 0D to 3D architectures. In this context, a zirconium tellurite (namely, ZrTe3O8) single crystal featuring a 3D distorted fluorite-type structure with a size of 35 × 32 × 21 mm3 was successfully harvested by the top-seeded solution growth (TSSG) technique. The X-ray diffraction rocking curve reflects that the crystallinity of the as-grown ZrTe3O8 crystal is quite perfect with a small full-width at half-maximum (fwhm) value (∼39 arcsec). The temperature dependence of the thermophysical properties of the ZrTe3O8 single crystal has been systematically analyzed. The ZrTe3O8 single crystal exhibits a wide transparency window, as the UV and IR absorption cutoff edges are respectively 278 and 7788 nm. The refractive indices over the region from the visible to the near-IR have been determined and manifested relatively large values of 2.0889-2.0370 over a wavelength range of 632.8-1553 nm. Furthermore, the fundamental physical characteristics of the ZrTe3O8 single crystal associated with its distinctive 3D framework structure have been evaluated with density functional theory (DFT) calculations.

Gene expression and hormone secretion profile of urotensin I associated with osmotic challenge in caudal neurosecretory system of the euryhaline flounder, Platichthys flesus.

The caudal neurosecretory system (CNSS) is a part of stress response system, a neuroendocrine structure unique to fish. To gain a better understanding of the physiological roles of CNSS in fluid homeostasis, we characterized the tissue distribution of urotensin I (UI) expression in European flounder (Platichthys flesus), analyzed the effect chronic exposure to seawater (SW) or freshwater (FW), transfer from SW to FW, and reverse transfer on mRNA levels of UI, L-type Ca2+ channels and Ca-activated K+ channels transcripts in CNSS. The tissue distribution demonstrated that the CNSS is dominant sites of UI expression, and UI mRNA level in fore brain appeared greater than other non-CNSS tissues. There were no consistent differences in CNSS UI expression or urophysis UI content between SW- and FW-adapted fish in July and September. After transfer from SW to FW, at 8 h CNSS UI expression was significantly increased, but urophysis UI content was no significantly changes. At 24 h transfer from SW to FW, expression of CNSS UI was no apparent change and urophysis UI content was reduced. At 8 h and 24 h after transfer from FW to SW UI expression and urophysis UI content was no significantly effect. The expression of bursting dependent L-type Ca2+ channels and Ca-activated K+ channels in SW-adapted fish significantly decreased compared to those in FW-adapted. However, there were no differences in transfer from SW to FW or from FW to SW at 8 h and 24 h. Thus, these results suggest CNSS UI acts as a modulator in response to osmotic stress and plays important roles in the body fluid homeostasis.

Rational Design of a LiNbO3-like Nonlinear Optical Crystal, Li2ZrTeO6, with High Laser-Damage Threshold and Wide Mid-IR Transparency Window.

With existing and emerging technologies urgently demanding the expansion of the laser wavelengths, high-performance nonlinear optical (NLO) crystals are becoming indispensable. Here, a prospective NLO crystal, Li2ZrTeO6, is rationally designed by the element substitution of Nb for Zr and Te from LiNbO3, which has been recognized as one of the most commercial NLO crystals. Li2ZrTeO6 with R3 symmetry inherits the structural merits of LiNbO3 (space group R3 c) and thus meets the requirements for NLO applications, including noncentrosymmetric crystal structure, moderate birefringence, and phase-matchability. Moreover, it can be exploited to achieve more outstanding optical damage resistant behavior (>1.3 GW cm-2), exceeding 22 times that of LiNbO3, which is more suitable for high-energy laser applications. Notably, this compound displays the widest IR absorption edge (7.4 µm) among all of the noncentrosymmetric tellurates reported so far. These excellent attributes suggest that Li2ZrTeO6 is a promising candidate for providing high NLO performance. The substitution of Nb for Zr and Te from LiNbO3 demonstrates a viable strategy toward the rational design of NLO crystals with anticipated properties.

Modulatory effect of glutamate GluR2 receptor on the caudal neurosecretory Dahlgren cells of the olive flounder, Paralichthys olivaceus.

A neuromodulatory role for glutamate has been reported for magnocellular neuroendocrine cells in mammalian hypothalamus. We examined the potential role of glutamate as a local intercellular messenger in the neuroendocrine Dahlgren cell population of the caudal neurosecretory system (CNSS) in the euryhaline flounder Paralichthys olivaceus. In pharmacological experiments in vitro, glutamate (Glu) caused an increase in electrical activity of Dahlgren cells, recruitment of previously silent cells, together with a greater proportion of cells showing phasic (irregular) activity. The glutamate substrate, glutamine (Gln), led to increased firing frequency, cell recruitment and enhanced bursting activity. The glutamate effect was not blocked by the N-methyl-D-aspartate (NMDA) receptor antagonist MK-801, or the GluR1/GluR3 (AMPA) receptor antagonist IEm1795-2HBr, but was blocked by the broad-spectrum α-amino-3-hydroxy- 5- methyl-4-isoxazo-lepropionic acid (AMPA) receptor antagonist ZK200775. Our transcriptome sequencing study revealed three AMPA receptor (GluR1, GluR2 and GluR3) in the olive flounder CNSS. Quantitative RT-PCR revealed that GluR2 receptor mRNA expression was significant increased following dose-dependent superfusion with glutamate in the CNSS. GluR1 and GluR3 receptor mRNA expression were decreased following superfusion with glutamate. L-type Ca2+ channel mRNA expression had a significant dose-dependent decrease following superfusion with glutamate, compared to the control. In the salinity challenge experiment, acute transfer from SW to FW, GluR2 receptor mRNA expression was significantly higher than the control at 2 h. These findings suggest that GluR2 is one of the mechanisms which can medicate glutamate action within the CNSS, enhancing electrical activity and hence secretory output.

Promoter methylation and Hoxd4 regulate UII mRNA tissue-specific expression in olive flounder (paralichthys olivaceus).

The peptide urotensin II (UII) mediates multiple physiology effects in mammals and fishes, and UII expression shows a tissue-specific pattern. However the mechanism is still unknown. In the present study high level of UII mRNA was detected in the caudal neurosecretory system (CNSS) of the olive flounder when compared to other tissues. We examined whether epigenetic mechanisms of DNA methylation are involved in UII gene expression. Methylation DNA immune precipitation (MeDIP) assay showed low methylation of UII promoter in CNSS tissue compared with muscle and spinal cord. Methylation of UII promoter was further assessed through bisulphate sequencing analysis. Low level methylation (31%) in CpG island of UII promoter was detected in CNSS tissue, while methylation status in muscle and spinal cord was 89% and 91%, respectively. In addition, high conserved sites of Hoxd4 in UII promoter were found. Activation of Hoxd4 mRNA using transretinoic acid (RA) resulted in 18-fold increase of UII mRNA expression in CNSS and high locomotor activity in medaka, confirming that Hoxd4 is also involved in UII gene transcriptional regulation. Taken together, our data provide the first evidence of the epigenetic mechanism of promoter methylation in transcriptional regulation of UII expression in a tissue-specific manner, and Hoxd4 may also participate in UII gene transcription in flounder.

Modulatory effect of dopamine receptor 5 on the neurosecretory Dahlgren cells of the olive flounder, Paralichthys olivaceus.

A neuromodulatory role for dopamine has been reported for magnocellular neuroendocrine cells in the mammalian hypothalamus. We examined its potential role as a local intercellular messenger in the neuroendocrine Dahlgren cell population of the caudal neurosecretory system (CNSS) of the euryhaline flounder Paralichthys olivaceus. In vitro application of dopamine (DA) caused an increase in electrical activity (firing frequency, recorded extracellularly) of Dahlgren cells, recruitment of previously silent cells, together with a greater proportion of cells showing phasic (irregular) activity. The dopamine precursor, levodopa (L-DOPA), also increased firing frequency, cell recruitment and enhanced bursting and tonic activity. The effect of dopamine was blocked by the D1, D5 receptor antagonist SCH23390, but not by the D2, D3, D4 receptor antagonist amisulpride. Transcriptome sequencing revealed that all DA receptors (D1, D2, D3, D4, and D5) were present in the flounder CNSS. However, quantitative RT-PCR revealed that D5 receptor mRNA expression was significantly increased in the CNSS following dopamine superfusion. These findings suggest that dopamine may modulate CNSS activity in vivo, and therefore neurosecretory output, through D5 receptors.

Quantitative proteomic reveals the dynamic of protein profile during final oocyte maturation in zebrafish.

The molecular mechanisms underlying final oocyte maturation in zebrafish (Danio rerio) remain poorly understood. The present study aimed to employ iTRAQ approach for a comprehensive characterization of during zebrafish oocyte maturation proteome and for comparison between fully-grow immature and mature oocytes prior to ovulation. A total of 1568 proteins were identified, which was representing the largest zebrafish isolated oocytes proteome dataset to date. Differential expression analysis revealed 190 proteins significantly changes between immature and mature oocytes, which 136 proteins were up-regulated and 54 proteins were down-regulated in mature oocytes comparison with immature oocytes. Functional analysis revealed that these differential proteins were mostly involved in cellular response to estrogen stimulus, cellular components, extracellular region, and enzyme regulator activity, etc. The revealed differentially changes in protein expression patterns associated with oocyte maturation suggest that several of the examined proteins, such as vitellogenin(Vtg3), protein S100(S100A10), 17-beta hydroxysteroid dehydrogenase(HSD17B1), pentaxin, zona pellucida (ZP3.2), elongation factor1-alpha, caluemnin B, and 14-3-3 protein may play a specific role during zebrafish final oocyte maturation. These data will provide powerful information for understanding the molecular mechanism underlying zebrafish oocyte maturation, and these proteins may potentially act as markers to predict control oocyte maturation of zebrafish oocytes.

Dynamic responses of prolactin, growth hormone and their receptors to hyposmotic acclimation in the olive flounder Paralichthys olivaceus.

Prolactin (PRL) and growth hormone (GH) play important roles in regulating salt and water balance through osmoregulatory organs in vertebrates. The aim of this study was to investigate the dynamic changes of GH/PRL hormone gene expressions in the pituitary gland and their receptors in gill and kidney, as well as the plasma osmolality when the olive flounder fish Paralichthys olivaceus were acclimated in freshwater (FW) conditions. After transfer from seawater (SW) to freshwater (FW), the osmolality of FW-adaption fish reached the lowest level at 1d which rose slightly afterwards. However, the hormone gene expression of PRL increased from 2d, reaching its peak at 5d, and then decreased at 14d. At this time, the value was still significantly higher than the control, showing a similar trend to the plasma hormone PRL. In contrast, the pituitary mRNA level of GH significantly decreased at 1d and then returned to normal levels. The mRNA levels of PRL receptor (PRLR) in both gill and kidney displayed a similar trend to the pituitary PRL. We also observed the synchronous expression trend of the renal PRLR with pituitary PRL (5d) and the asynchronous expression peaks between branchial (8d) and renal PRLR (5d). Significant responses of GH and its receptor (GHR) in both gill and kidney during the FW-acclimation were not observed. Nevertheless, the gene expression of GH receptor variant (GHR-V) in both gill and kidney declined at 2d, indicating unknown osmoregulatory functions of GHR-V. Collectively, our results provided more insights of the PRL, GH and their corresponding receptors in modulating osmoregulatory responses, representing an important aspect of FW-acclimation in flounder fish.

Molecular characterisation and expression of parathyroid hormone-related protein in the caudal neurosecretory system of the euryhaline flounder, Platichthys flesus.

Parathyroid hormone-related protein (PTHrP) is a hypercalcemic factor in fish, but the source of circulating PTHrP remains unclear. In this study investigation of the caudal neurosecretory system (CNSS), considered one of major sources of PTHrP in fish, provided valuable insights into this regulatory system. We report pthrpa and pthrpb gene cloning, characterization, expression, and responses to low salinity and hypocalcemia challenge in flounder. The pthrpa and pthrpb precursors, isolated from a European flounder CNSS library, consist of 166 and 192 amino acid residues, respectively, with an overall homology of approximately 59.2%. Both precursors contain a signal peptide and a mature peptide with cleavage and amidation sites. The flounder PTHrPA and PTHrPB peptides share only 41% sequence identity with human PTHrPA. Quantitative PCR analysis demonstrated that the bone and bladder, are respectively major sites of pthrpa and pthrpb expression in flounder. Urophysectomy confirmed the CNSS as a likely contributor to circulating PTHrP peptides. There were no significant differences in CNSS pthrpa and pthrpb mRNA expression or plasma PTHrP levels between seawater (SW) and freshwater (FW)-adapted fish, though plasma total calcium concentrations were higher in FW animals. The intraperitonial administration of EGTA rapidly induced hypocalcemia and concomitant elevation in plasma PTHrP accompanied by increases in both pthrpa and pthrpb expression in the CNSS. Together, these findings support an evolutionary conserved role for PTHrP in the endocrine regulation of calcium.

Antioxidant response of the hard shelled mussel Mytilus coruscus exposed to reduced pH and oxygen concentration.

Ocean acidification (OA) and hypoxic events are increasing worldwide problems, their interactive effects have not been well clarified, although their co-occurrence is prevalent. The East China Sea (the Yangtze River estuary area) suffers from not only coastal hypoxia but also pH fluctuation, representing an ideal study site to explore the combined effect of OA and hypoxia on marine bivalves. We experimentally evaluated the antioxidant response of the mussel Mytilus coruscus exposed to three pH levels (8.1, 7.7 and 7.3) at two dissolved oxygen (DO) levels (2.0mgL-1 and 6.0mgL-1) for 72h. Activities of superoxide dismutase, catalase, glutathione peroxidase, acid phosphatase, and alkaline phosphatase and levels of malondialdehyde were measured in gills and hemolymph. All enzymatic activities in hemolymph and gills followed a similar pattern throughout the experiment duration. Generally, low DO showed greater effects on enzyme activities than elevated CO2. Significant interactions between DO, pH and time were only observed at superoxide dismutase and catalase in both tissues. PCA revealed positive relationships between most enzyme activities in both gills and hemolymph with the exception of alkaline phosphatase activity and the level of malondialdehyde in the hemolymph. Overall, our results suggested that decreased pH and low DO induced similar antioxidant responses in the hard shelled mussel, and showed an additive effect on most enzyme activities. The evaluation of multiple environmental stressors, a more realistic scenario than single ones, is crucial to predict the effect of future global changes on coastal species and our results supply some insights on the potential combined effects of reduced pH and DO on marine bivalves.