Search

Chile is the second-largest producer of Atlantic salmon in the world, and the Chilean salmon production accounts for 27% of the world’s production. One important step of the productive cycle in freshwater is the smoltification process that prepares the fish for the marine life stage. This review describes the biological basis of smoltification in Atlantic salmon, with particular attention on branchial osmoregulatory adaptations. We also discuss some of the infectious diseases and problems in smoltification (two of the main causes of losses in Chilean aquaculture) that could be related from a physiological point of view.

Studying the evolutionary history of gene families is a challenging and exciting task with a wide range of implications. In addition to exploring fundamental questions about the origin and evolution of genes, disentangling their evolution is also critical to those who do functional/structural studies to allow a deeper and more precise interpretation of their results in an evolutionary context. The sirtuin gene family is a group of genes that are involved in a variety of biological functions mostly related to aging. Their duplicative history is an open question, as well as the definition of the repertoire of sirtuin genes among vertebrates. Our results show a well-resolved phylogeny that represents an improvement in our understanding of the duplicative history of the sirtuin gene family. We identified a new sirtuin gene family member (SIRT3.2) that was apparently lost in the last common ancestor of amniotes but retained in all other groups of jawed vertebrates. According to our experimental analyses, elephant shark SIRT3.2 protein is located in mitochondria, the overexpression of which leads to an increase in cellular levels of ATP. Moreover,in vitroanalysis demonstrated it has deacetylase activity being modulated in a similar way to mammalian SIRT3. Our results indicate that there are at least eight sirtuin paralogs among vertebrates and that all of them can be traced back to the last common ancestor of the group that existed between 676 and 615 millions of years ago.

The TAR DNA Binding Protein (TARDBP) gene has become relevant after the discovery of its several pathogenic mutations. The lack of evolutionary history is in contrast to the amount of studies found in the literature. This study investigated the evolutionary dynamics associated with the retrotransposition of the TARDBP gene in primates. We identified novel retropseudogenes that likely originated in the ancestors of anthropoids, catarrhines, and lemuriformes, i.e. the strepsirrhine clade that inhabit Madagascar. We also found species-specific retropseudogenes in the Philippine tarsier, Bolivian squirrel monkey, capuchin monkey and vervet. The identification of a retropseudocopy of the TARDBP gene overlapping a lncRNA that is potentially expressed opens a new avenue to investigate TARDBP gene regulation, especially in the context of TARDBP associated pathologies.

Pathogen interactions with cultured fish populations are well studied, but their effects on native fishes have not been characterized. In Chile, the disease caused by bacterial species Piscirickettsia salmonis represents one of the main issues and is considered to be one of the important pathogens in the field of aquaculture. They have been found to infect native fish. Therefore, it is necessary to understand the impact of P. salmonis on native species of local commercial value, as well as the potential impact associated with the emergence of antibiotic-resistant strains of P. salmonis. Due to this purpose, the native fish Eleginops maclovinus was used in our study. Fish were randomly distributed in tanks and intraperitoneally inoculated with two strains of P. salmonis. No mortality was recorded during the experiment. Cortisol, glucose and total α-amino acid levels increased in fish injected with AUSTRAL-005 strain compared to sham-injected and LF-89-inoculated fish. Moreover, results showed an increase in the activity of carbohydrates and lipids metabolism in liver; and an increase in the carbohydrates, lipids and total α-amino acid metabolism in muscle after injection with AUSTRAL-005. Our results suggest that P. salmonis modulates the physiology of E. maclovinus and the physiological impact increase in the presence of the antibiotic-resistant strain AUSTRAL-005.

Pseudomyxoma peritonei (PMP) is a disease surrounded by misunderstanding and controversies. Knowledge about the etymology of pseudomyxoma is useful to remove the ambiguity around that term. The word pseudomyxoma derives from pseudomucin, a type of mucin. PMP was first described in a case of a woman alleged to have a ruptured pseudomucinous cystadenoma of the ovary, a term that has disappeared from today's classifications of cystic ovarian neoplasms. It is known today that in the majority of cases, the origin for PMP is an appendiceal neoplasm, often of low histological grade. Currently, ovarian tumors are wrongly being considered a significant recognized etiology of PMP. PMP classification continues to be under discussion, and experts' panels strive for consensus. Malignancy is also under discussion, and it is shown in this review that there is a long-standing historical reason for that. Surgery is the main tool in the treatment armamentarium for PMP, and the only therapy with potential curative option.

The objective of this study was to investigate early postmortem (0.5 h) gene expression in beef Longissimus thoracis (LT) muscles from carcasses with NORMAL ( 5.9) ultimate pH (pHu). A total of 53 transcripts were differentially expressed (P-value

The aquaculture industry has many problems associated with bacteria, viruses, and parasites, and the stress caused by these infections can modulate the physiological response in fish. In Chile, the sea lice Caligus rogercresseyi constitutes a major problem affecting the Chilean salmonid industry, having a strong negative effect on salmon production. For this reason, the aim of this study was to investigate the neuroendocrine and stress response of the most commonly cultured salmonid species in Chile, the Atlantic salmon (Salmo salar) and the Coho salmon (Oncorynchus kisutch), which were experimentally infested with the sea lice Caligus rogercresseyi. We analyzed the monoamine response in the telencephalon, optic tectum, and hypothalamus, as well as cortisol levels in plasma. In the liver and muscle we analyzed the cellular response by measuring heat shock proteins (60, 70, 90) and glucocorticoid mRNA transcription. After 14 days post infection C. rogercresseyi infestation modified the neuroendocrine and stress response in Atlantic and Coho salmon, with Coho salmon presenting a faster and higher response than the Atlantic salmon.

Variations in the mRNA expression of hepatic and muscle genes that are related to calcium signaling were analyzed by real-time qPCR in farmed Atlantic salmon (Salmo salar L. 1758) to determine changes in expression between parr and smolt stages. These organs were selected due to their close relationship with calcium signaling and metabolism (e.g., glycolysis, oxidative phosphorylation, muscle contraction). Differential expression between smolt and parr specimens and between organs was observed. Compared to parr specimens, smolts exhibited upregulated expression of the calcitonin receptor precursor, calcitonin receptor, calcitonin isoform, parathyroid hormone, and calmodulin in the liver. This pattern was inverse in muscle, with the exception of calmodulin, which was significantly upregulated in smolts compared to parr. Additionally, plasma calcium was decreased in the smolt condition. This study is the first to characterize the expression pattern of calcium signaling-related genes in the liver and muscle of parr and smolt S. salar. However, further functional studies are required to obtain a wider understanding about the physiological changes that accompany the productive conditions during smoltification.

Piscirickettsiosis caused by Piscirickettsia salmonis constitutes one of the main problems in farmed salmonid and marine fishes. The objective of this study was to evaluate the modulation of genes involved in the oxidative stress in the liver and muscle of Salmo salar challenge with low dosage of P. salmonis. The treatment (in duplicate) were as follows: Control injection (culture medium) and P. salmonis injection (1 × 102 PFU/mL) with sampling (liver and muscle) at several time-points during the 42-days experimental period (dpi). In liver, the gene expression of superoxide dismutase (SOD) and acetylcholinesterase (AChE) had differences with the control group only at 7 dpi, compared with glutathione-S-transferase (GST) and heat shock protein 70 (HSP70) that presented increases at 7 and 21 dpi. The glutathione peroxidase (GPx) and catalase (CAT) mRNAs were elevated at 13 and 21 dpi, respectively. While glutathione reductase (GR) and cytochrome P450 (P450) did not show variations in their expression during the experimental course. In muscle, the expression of CAT and AChE was higher than in the control condition at 2 and 42 dpi, respectively. While the number of transcripts SOD, GPx, GR, GST, P450 and HSP70 showed increases at 7- and 42-days post injection. The results suggest a transcriptional activation of genes involved in oxidative stress in both liver and muscle, with expression profiles that were tissue-specific and dependent on the time. This is the first study that reveals the transcriptional participation of all these genes associated with oxidative stress in response to the injection of P. salmonis.

Gurltia paralysans is an angio-neurotropic metastrongyloid nematode that infects domestic and wild cats, invading the veins of the subarachnoid space of the spinal cord and mainly causing progressive paralysis of the pelvic limbs. The definitive diagnosis of feline gurltiosis can only be achieved by post-mortem examination that reveals the presence of the nematode in the spinal cord vein vasculature. An early diagnosis with conclusive results is required since laboratory and imaging findings are not sufficient. Therefore, the purpose of this study was to detect the presence of G. paralysans, via semi-nested PCR, in samples of cerebrospinal fluid (CSF) and the sera of domestic cats naturally infected with the parasite. A total of 12 cats with a diagnosis suggestive of feline gurltiosis were selected, and they underwent a complete neurological and imaging examination. DNA samples were analysed by semi-nested PCR, with universal (AaGp28Sa1/AaGp28Ss1) and specific (Gp28Sa3/Aa28Ss2) primers, for G. paralysans (G. paralysans 18S rRNA gene, partial sequence, ITS 1, 5.8S rRNA gene, and ITS 2, complete sequence, and 28S rRNA gene, partial sequence) and Aelurostrongylus abstrusus, obtaining amplifications of 356 and 300 bp, which indicated the presence or absence of nematode DNA, respectively. The presence of G. paralysans was detected in the CSF of four out of nine cats, and the sera of seven out of seven cats. In the sera analysis of five out of seven cats, a mixed infection with A. abstrusus was found, despite no alterations of the respiratory tract being observed during the necropsies. It is proposed that serum samples could be more effective than CSF in detecting the parasite by PCR analysis. Sequencing analysis showed high percentages of identity with G. paralysans, which indicated the feasibility of detection and the sensitivity/specificity of the method used, suggesting the implementation of semi-nested PCR as a routine diagnostic test for early and timely detection of feline gurltiosis.

BK potassium channels are implicated in a variety of physiological processes and they have been characterized in many species from Drosophila to humans although detailed studies about their properties and functions in the osmoregulation of teleosts are still lacking. In this study, we have cloned 911 bp corresponding to a partial Salmo salar BK channel coding sequence with high level of homology to BK channel sequences from other teleosts. Also, we have followed the BK gene expression pattern in gills of Atlantic salmon during brackish water (BW) acclimation. Interestingly, transfer from freshwater to BW induced an upregulation in mRNA BK channel expression in gills 7 days after transfer to BW, followed by a decrease in the expression. These results are the first describing mRNA changes of BK channel in gills of Atlantic salmon during BW acclimation, and they could suggest a possible role for BK channel during this adaptive response. Additionally, our results could position BK potassium channels expressed in the gills of Salmo salar as new candidates to be a marker of smoltification process, which may complement, and possibly outperform NKA activity measurement, the current gold standard of the Chilean industry.

Smoltification under intensive aquaculture conditions involves important physiological changes to intestinal structures that facilitate acclimation to hyperosmotic environments. This study evaluated if genic expression of primary contributors during the smoltification process in the intestine are in synchrony with gill Na+/K+-ATPase activity. Discordance was found between gill Na+/K+-ATPase activity levels and intestinal genic expressions. In particular, the intestine showed increased mRNA expression for prolactin, a hormone that participates in hypoosmotic environments, and in mRNA expression of the growth hormone, which mediates the hydromineral balance in hyperosmotic environments. These findings indicate a lack of sufficient intestinal maturation in the fish at the moment when they were transferred to seawater environments in the normal aquaculture practices. This intestinal immaturity is a possible explanation for the high rates of desadapted individuals in the seawater phase in Chilean salmon aquaculture.

Failure of pancreaticoenteric anastomos is a major cause of morbidity after pancreaticoduodenectomy. Factors that contribute to the poor outcome of this anastomosis are soft glandular texture, and small pancreatic duct diameter. Invaginating pancreatogastrostomy has been deemed superior to pancreaticojejuostomy when those risk factors are present. Nevertheless, the suture between gastric mucosa and pancreas is often a bothersome step and has a potential contaminating load to the surgical field. Herein, a simplified method for suturing the pancreatic stump into the stomach is shown, avoiding the complexity of the suture and minimizing the manipulation into the gastric lumen.

Although Caligus rogercresseyi negatively impacts Chilean salmon farming, the metabolic effects of infection by this sea louse have never been completely characterized. Therefore, this study analyzed lactate responses in the plasma, as well as the liver/muscle lactate dehydrogenase (LDH) activity and gene expression, in Salmo salar and Oncorhynchus kisutch infested by C. rogercresseyi. The lactate responses of Atlantic and Coho salmon were modified by the ectoparasite. Both salmon species showed increasing in plasma levels, whereas enzymatic activity increased in the muscle but decreased in the liver. Gene expression was overexpressed in both Coho salmon tissues but only in the liver for Atlantic salmon. These results suggest that salmonids need more energy to adapt to infection, resulting in increased gene expression, plasma levels, and enzyme activity in the muscles. The responses differed between both salmon species and over the course of infection, suggesting potential species-specific responses to sea-lice infection.

Recombination in alphaherpesviruses was first described more than sixty years ago. Since then, different techniques have been used to detect recombination in natural (field) and experimental settings. Over the last ten years, next-generation sequencing (NGS) technologies and bioinformatic analyses have greatly increased the accuracy of recombination detection, particularly in field settings, thus contributing greatly to the study of natural alphaherpesvirus recombination in both human and veterinary medicine. Such studies have highlighted the important role that natural recombination plays in the evolution of many alphaherpesviruses. These studies have also shown that recombination can be a safety concern for attenuated alphaherpesvirus vaccines, particularly in veterinary medicine where such vaccines are used extensively, but also potentially in human medicine where attenuated varicella zoster virus vaccines are in use. This review focuses on the contributions that NGS and sequence analysis have made over the last ten years to our understanding of recombination in mammalian and avian alphaherpesviruses, with particular focus on attenuated live vaccine use.

Myelosupression resulting from chemotherapy has been widely described in veterinary medicine; however, there is limited information relating to alterations in neutrophil function after chemotherapy in dogs with cancer. The aim of this study was to determine the non-proliferative effects of vincristine, carboplatin, and cisplatin on canine neutrophils by evaluating activation of oxidative and non-oxidative responses. Neutrophils were isolated from venous blood. Levels of reactive oxygen species (ROS) and metalloproteinase 9 (MMP-9) were measured in vitro during neutrophil exposure to these chemotherapeutic agents for 15 min followed by stimulation with platelet activating factor (PAF). ROS production was detected via luminescence, and MMP- 9 liberation was determined by zymography. The chemotherapeutic agents caused an increase in PAF-induced ROS production, but no change in the non-oxidative response was observed. These results suggest that these chemotherapeutic agents may act as priming agents by increasing the oxidative response. These effects could be beneficial for dogs with cancer by supporting their immune systems; however, excessive ROS liberation has been associated with inflammation, neutrophil-mediated cell injury, carcinogenesis, and metastasis. Clinical studies are necessary to evaluate the significance of these findings.

The aim of this study was to examine the effects of Flavobacterium psychrophilum, a pathogen that is economically important in the aquaculture sector, on the neuroendocrine response of Oncorhynchus mykiss during a time course experiment with sampling at 0.5, 1, 2, 6, 10, and 30 days post injection (dpi). In the brain, serotonin (5HT) content increased in the infected group at all the measured time points, a similar pattern was observed for 5-hydroxyindole-3-acetic acid (5HIAA). Infected fish presented an increase in brain dopamine levels on day 0.5 and 1 dpi. A non-significant variation in noradrenaline levels was observed on all treatment days. Foregut 5-HT and 5-HIAA content in the infected group presented the highest 5-HT concentrations with 248.6 and 983.5 ng/g tissue at 0.5 dpi respectively. Midgut 5-HT and 5-HIAA levels presented the highest 5-HT concentrations, 486.9 ng/g tissue and 1006.4 ng/g tissue respectively, at the beginning of the experiment (0.5 dpi). 5-HT levels in the hindgut presented the highest concentrations with 233.9 ng/g tissue at 0.5 dpi, while 5-HIAA presented the highest concentrations, 690.5 ng/g tissue, at the same time point. After injection with F. psychrophilum the neuroendocrine response in rainbow trout was tissue dependent. Brain levels of 5HT and 5HIIA indicate that the neuroendocrine response increased together with dopamine following intramuscular infection. These increases are in line with reports from other authors, indicating an early response of catecholamines as neurotransmitters to stressful stimulus. In addition the intestinal response was also increased, implying that there could be a possible relationship between the serotonergic system at the intestinal level and the immune system.

Simple Summary Within innate immunity, polymorphonuclear neutrophils (PMN) are the most abundant leukocyte population. Alongside PMN, monocytes, eosinophils, and basophils are also known to exist. All of them can release extracellular traps (ETs), a complex web-like structure composed of chromatin decorated with nuclear histones, granular enzymes, peptides, and proteins, to firmly entrap invasive pathogens, thereby slowing dissemination and helping to develop proper immune responses against bacteria, fungi, viruses, and parasites. Here, we showed for the first time that Atlantic salmon-derived PMN released ETs-like structures in vitro, in response to highly pathogenic facultative intracellular rickettsial bacteria Piscirickettsia salmonis. The release of ET-like structures from PMN could be a new alternative to improve farmed salmon’s defense against pathogens. Abstract Extracellular traps (ETs) are webs of DNA, citrullinated histones, anti-microbial peptides, and proteins that were not previously reported in Atlantic salmon (Salmo salar). ETs are mainly released from polymorphonuclear neutrophils (PMN) and are considered a novel PMN-derived effector mechanism against different invasive pathogens. Here, we showed that Atlantic salmon-derived PMN released ETs-like structures in vitro in response to highly pathogenic facultative intracellular rickettsial bacteria Piscirickettsia salmonis. PMN were isolated from pre-smolt Atlantic salmon and stimulated in vitro with oleic acid and P. salmonis. Extracellular DNA was measured using the PicoGreen™ dye, while immunofluorescence image analysis was used to confirm the classical components of salmonid-extruded ETs. Future studies are required to better understand the role of Atlantic salmon-derived ETs orchestrating innate/adaptive immunity and the knowledge on regulation pathways involved in this cell death process. Thus, comprehension of salmonid-derived ETs against P. salmonis might represent novel alternative strategies to improve host innate defense mechanisms of farmed salmon against closely related rickettsial bacteria, as a complement to disease prevention and control strategies.

Francisella noatunensis subsp. noatunensis is the responsible agent of Francisellosis, a bacterial disease that affects an important amount of aquatic farmed species. Eleginops maclovinus is a fish that cohabits with salmonids cages in Chile and can also act as a vector of this bacterial disease. In the present study, we evaluated calcium metabolism in the liver of E. maclovinus injected intraperitoneally with different doses of F. noatunensis subsp. noatunensis (low 1.5 × 101, medium 1.5 × 105 and high doses 1.5 × 1010 cells/μL). Fish were sampled at 1, 3, 7, 14, 21 and 28 days post injection (dpi). No mortalities nor clinical signs were observed. Plasma calcium levels were higher in the high doses group of F. noatunensis subsp. noatunensis at day 7 and 14 compared to the control group (fish injected with bacterial medium alone). Hypercalcemic factors increased at day 14 and 21 for the medium and low dose (parathyroid hormone-related protein precursor), while vitamin D3 receptor increased its expression at times 1, 3 and 7 for the low dose. On the other hand, hypocalcemic factors such as calcitonin receptor and stanniocalcin increased its expression at time 7 and 14, respectively. Calmodulin involved in calcium storage decreased its expression during all experimental days in fish subjected to high bacterial dose. Proteins involved in calcium transport, such as L-type voltage-gated calcium channel and trpv5 increased their transcription at day 1 and 14, compared to calcium sensing-receptor and plasma membrane Ca2 +- ATPase that showed peak expression at times 14 and 28. The results suggest a clear alteration of calcium metabolism, mainly in high bacterial doses. This study provides new knowledge about the calcium metabolism in fish infected with bacteria.

Eleginops maclovinus is a native species with potential for Chilean aquaculture. However, its optimal stocking density and the impact on its physiology is unknown. The aim of this study is to determine the optimal stocking density range by evaluating parameters of growth, stress, intermediary metabolism, and osmoregulation. For this, 48 immature male fish (201.60 ± 32 g; 29.05 ± 1.45 cm) were randomly assigned to four stocking densities (3, 6, 12 and 24 kg/m3) and maintained for 60 days. No mortality was recorded, optimal growth was observed at a density of 3–6 kg/m3 and decreased with a higher stocking density. In plasma, glucose and protein levels were not affected by stocking density, while triglycerides, lactate, total amino acids, cortisol, and osmolality showed a significant decrease at a higher stocking density. Enzymatic activity of branchial and renal NKA, in addition to water content in muscle, increased at a greater stocking density, while intestinal NKA showed non-significant differences. The metabolism of carbohydrates, triglycerides, and amino acids/protein in liver, gill, kidney, and muscle was significantly affected by stocking density and showed significant tissue-specific responses. Glycogen levels increased at a higher stocking density in gills, kidney, and muscle, suggesting that glucose plays an important role against chronic stress produced by stocking density. The GH/IGF-1 axis and HSP70 showed significantly lower levels of mRNA transcript at the high density of 24 kg/m3. In conclusion, we suggest that stocking density affects the physiology of Eleginops maclovinus, activating the stress system and mobilizing energy substrates in a tissue-specific manner. Additionally, our data suggests that male E. maclovinus should be cultivated in the range of 3–12 kg/m3, avoiding the high density of 24 kg/m3.

Eleginops maclovinus is a sub-Antarctic fish endemic to the South American coast and is a monotypic species of the Eleginopidae family. A scarce amount of scientific information exists regarding the effects of temperature on E. maclovinus physiology. In this study, E. maclovinus specimens were acclimated for 2 weeks at 10 °C (control), 14, and 18 °C. Posterior assessments were performed for the intermediary metabolism of carbohydrates, lipids, and amino acids (in the plasma, liver, gills, and kidney) and on the osmoregulatory capacity [plasma osmolality and gill, kidney, and foregut Na+, K+-ATPase (NKA) activities]. In the plasma, only lactate and total amino acid levels were affected by the temperature. NKA activity presented a linear relationship with the increased temperatures. The liver exhibited no change in the carbohydrates metabolism, but the glycogen/glucose metabolite levels did differ. Amino acid metabolism showed a direct relationship between temperature and GDH and Asp-AT activities, and an inverse relationship between temperature and Ala-AT activity. Lipid metabolism was similar to Ala-AT. In the gills, carbohydrate metabolism (G6PDH and HK activities) presented a direct relationship with the increased temperatures. In the kidney, carbohydrate metabolism (only G6PDH activity) was greater at higher temperatures, and amino acid metabolism (GDH activity) showed a direct relationship with temperature. Lipid metabolism (G3PDH activity) was greatest at 14 °C, presenting significant differences compared to 10 °C. The obtained results suggest that E. maclovinus can acclimate to the projected thermal increase of climate change, with a reorganization of the intermediary metabolism components being tissue-dependent and osmoregulatory effects being affected by NKA activity and greater rates of nitrogenous waste extrusion.

Voltage-gated ion channels are the molecular determinants of cellular excitability. This group of ion channels is one of the most important pharmacological targets in excitable tissues such as nervous system, cardiac and skeletal muscle. Moreover, voltage-gated ion channels are expressed in non-excitable cells, where they mediate key cellular functions through intracellular biochemical mechanisms rather than rapid electrical signaling. This review aims at illustrating the pharmacological impact of these ion channels, highlighting in particular the structural details and physiological functions of two of them - the high conductance voltage- and Ca(2+)-gated K(+) (BK) channels and voltage-gated proton (Hv1) channels- in non-excitable cells. BK channels have been implicated in a variety of physiological processes ranging from regulation of smooth muscle tone to modulation of hormone and neurotransmitter release. Interestingly, BK channels are also involved in modulating K(+) transport in the mammalian kidney and colon epithelium with a potential role in the hyperkalemic phenotype observed in patients with familial hyperkalemic hypertension type 2, and in the pathophysiology of hypertension. In addition, BK channels are responsible for resting and stimulated Ca(2+)-activated K(+) secretion in the distal colon. Hv1 channels have been detected in many cell types, including macrophages, blood cells, lung epithelia, skeletal muscle and microglia. These channels have a central role in the phagocytic system. In macrophages, Hv1 channels participate in the generation of reactive oxygen species in the respiratory burst during the process of phagocytosis.

The main role of voltage-gated proton channels (Hv1) is to extrude protons from the intracellular milieu when, mediated by different cellular processes, the H+ concentration increases. Hv1 are exquisitely selective for protons and their structure is homologous to the voltage sensing domain (VSD) of other voltage-gated ion channels like sodium, potassium, and calcium channels. In clear contrast to the classical voltage-dependent channels, Hv1 lacks a pore domain and thus permeation necessarily occurs through the voltage sensing domain. Hv1 channels are activated by depolarizing voltages, and increases in internal proton concentration. It has been proposed that local conformational changes of the transmembrane segment S4, driven by depolarization, trigger the molecular rearrangements that open Hv1. However, it is still unclear how the electromechanical coupling is achieved between the VSD and the potential pore, allowing the proton flux from the intracellular to the extracellular side. Here we provide a revised view of voltage activation in Hv1 channels, offering a comparative scenario with other voltage sensing channels domains.

Caligus rogercresseyi, a marine ectoparasite, causes notable economic losses for the Chilean salmonid industry. Nevertheless, the immunological responses of infected fish remain poorly understood, including proinflammatory cytokine generation and the respective modulatory effects of various cytokine receptors. This study evaluated mRNA expression of the NLRC5, major histocompatibility complex (MHC) class II, I-kappa-B-alpha, a regulatory that inhibits NF-kappa-B, and proinflammatory cytokines (IL-1&beta, and IL-18) in the liver and muscle of Atlantic salmon (Salmo salar) and Coho salmon (Oncorhynchus kisutch) during a time-course C. rogercresseyi infestation trial. All assessed mRNA were strongly regulated during infestation, but S. salar showed up-regulated expression, possibly accounting for the high infestation vulnerability of this salmonid. In conclusion, this work helps to understand the modulation of the expression of different transcripts involved over short periods of C. rogercresseyi infestation in two salmonid species (S. salar and O. kisutch).

The NOD-like receptors (NLRs) were recently identified as an intracellular pathogen recognition receptor family in vertebrates. While the immune system participation of NLRs has been characterized and analyzed in various mammalian models, few studies have considered NLRs in teleost species. Therefore, this study analyzed the Atlantic salmon (Salmo salar) NLRC5. Structurally, Atlantic salmon NLRC5 presented leucine-rich repeat subfamily genes. Phylogenetically, NLRC5 was moderately conserved between S. salar and other species. Real-time quantitative PCR revealed NLRC5 expression in almost all analyzed organs, with greatest expressions in the head kidney, spleen, and hindgut. Furthermore, NLRC5 gene expression decreased during smolt stage. These data suggest that NLRC5 participates in the Atlantic salmon immune response and is regulated, at least partly, by the smoltification process, suggesting that there is a depression of immune system from parr at smolt stage. This is the first report on the NLRC5 gene in salmonid smolts.

Caligus rogercresseyi sea lice negatively impact Chilean salmonid farming, but no complete characterization for the metabolic effects of Caligus infestation currently exists. Therefore, the aim of this study was to analyze the effects of C. rogercresseyi infestation on the metabolic responses of Salmo salar and Oncorhynchus kisutch. Energy metabolism responses to C. rogercresseyi were examined over a time-course infestation of both salmonid species. Plasma metabolite levels and enzymatic activities related to intermediate metabolisms of carbohydrate, amino acid, and lipid were evaluated in the liver and muscle. Plasma glucose levels changed in both salmonid species, increasing at 1–3 days post-infestation. In turn, triglyceride levels increased at days 3 and 7 in O. kisutch and S. salar, respectively, while protein and total α-amino acids increased in O. kisutch but decreased in S. salar during infestation. Amino acid intermediate catabolic metabolism in the liver and muscle of O. kisutch increased during infestation, indicating a higher use of the gluconeogenic pathway than S. salar. Lipid intermediate anabolic metabolism increased in O. kisutch liver, remained unchanged in S. salar liver, and increased 1 day post-infestation in the muscle of both salmonids. Liver and muscle carbohydrate intermediate anabolic metabolism in O. kisutch increased during infestation, suggesting that this species preferentially uses the glycogenolytic-glucolytic pathway, in contrast with S. salar. In conclusion, amino acid and carbohydrate catabolism enzymes in O. kisutch activated soon after initial sea lice infestation, which would allow this species to dispose of energy substrates earlier than S. salar. This physiological data contributes towards the ability of O. kisutch to more adeptly cope with the increased energy demand imposed by C. rogercresseyi infestation.

The BK channel is one of the most broadly expressed ion channels in mammals. In many tissues, the BK channel pore-forming α-subunit is associated to an auxiliary β-subunit that modulates the voltage- and Ca2+-dependent activation of the channel. Structural components present in β-subunits that are important for the physical association with the α-subunit are yet unknown. Here, we show through co-immunoprecipitation that the intracellular C-terminus, the second transmembrane domain (TM2) and the extracellular loop of the β2-subunit are dispensable for association with the α-subunit pointing transmembrane domain 1 (TM1) as responsible for the interaction. Indeed, the TOXCAT assay for transmembrane protein–protein interactions demonstrated for the first time that TM1 of the β2-subunit physically binds to the transmembrane S1 domain of the α-subunit. Structured summary of protein interactions: BK channel subunit alpha physically interacts with BK channel subunit beta-2 by anti tag coimmunoprecipitation ( View interaction )

This study describes de novo transcriptome sequencing and annotation analyses for the head kidney of the sub-Antarctic notothenioid fish Eleginops maclovinus, a sister group of the Antarctic notothenioid fish clade. Moreover, E. maclovinus is one of the most eurythermal and euryhaline representatives of the Notothenioidei suborder. RNA-seq data were generated by the 454 GS Junior system, resulting in 11,207 contigs that were then assembled by the Genomic Workbench CLC software. The transcriptome was annotated by BLASTing each sequence against the universal, non-redundant NCBI database (National Center for Biotechnology Information) using the AUSTRAL-omics computer cluster. A significant number of transcripts related to innate and adaptive immunity were found in the sequences, which could be used as references in future immunological studies in E. maclovinus.

A number of potassium channels including members of the KCNQ family and the Ca2+ activated IK and SK, but not BK, are strongly and reversibly regulated by small changes in cell volume. It has been argued that this general regulation is mediated through sensitivity to changes in membrane stretch. To test this hypothesis we have studied the regulation of KCNQ1 and BK channels after expression in Xenopus oocytes. Results from cell-attached patch clamp studies (∼50 μm2 macropatches) in oocytes expressing BK channels demonstrate that the macroscopic volume-insensitive BK current increases with increasing negative hydrostatic pressure (suction) applied to the pipette. Thus, at a pipette pressure of −5.0 ± 0.1 mmHg the increase amounted to 381 ± 146% (mean ±s.e.m., n= 6, P < 0.025). In contrast, in oocytes expressing the strongly volume-sensitive KCNQ1 channel, the current was not affected by membrane stretch. The results indicate that (1) activation of BK channels by local membrane stretch is not mimicked by membrane stress induced by cell swelling, and (2) activation of KCNQ1 channels by cell volume increase is not mediated by local tension in the cell membrane. We conclude that stretch and volume sensitivity can be considered two independent regulatory mechanisms.

The movement of ions across cell membranes is essential for a wide variety of fundamental physiological processes, including secretion, muscle contraction, and neuronal excitation. This movement is possible because of the presence in the cell membrane of a class of integral membrane proteins dubbed ion channels. Ion channels, thanks to the presence of aqueous pores in their structure, catalyze the passage of ions across the otherwise ion-impermeable lipid bilayer. Ion conduction across ion channels is highly regulated, and in the case of voltage-dependent K(+) channels, the molecular foundations of the voltage-dependent conformational changes leading to the their open (conducting) configuration have provided most of the driving force for research in ion channel biophysics since the pioneering work of Hodgkin and Huxley (Hodgkin, A. L., and Huxley, A. F. (1952) J. Physiol. 117, 500-544). The voltage-dependent K(+) channels are the prototypical voltage-gated channels and govern the resting membrane potential. They are responsible for returning the membrane potential to its resting state at the termination of each action potential in excitable membranes. The pore-forming subunits (alpha) of many voltage-dependent K(+) channels and modulatory beta-subunits exist in the membrane as one component of macromolecular complexes, able to integrate a myriad of cellular signals that regulate ion channel behavior. In this review, we have focused on the modulatory effects of beta-subunits on the voltage-dependent K(+) (Kv) channel and on the large conductance Ca(2+)- and voltage-dependent (BK(Ca)) channel.

Atlantic salmon (Salmo salar) and Rainbow trout (Oncorhynchus mykiss) are two fish species that spawn in fresh water (FW) and, during development, acclimate to seawater (SW) by secreting excess NaCl to the environment. The salmon industry measures Na+/K+ ATPase (NKA) activity as a molecular marker to determine the timing of smolt transfer from FW to SW. However, the lack of other accurate molecular markers of smoltification remains a major issue for the fish farming industry. The molecular mechanism of NaCl secretion in gills from SW-acclimated fish has a thermodynamic requirement to recycle K+ out of the cell via potassium channels therefore we hypothesised that potassium channel expression in gills may be a suitable candidate to monitor the smoltification process. In support of this hypothesis, we observed increased expression of BK potassium channel mRNA in gills from S. salar under conditions of high salinity (1.2%) compared to animals in FW. In this work, we designed a real-time PCR analysis in order to quantify mRNA levels of BK potassium channels in S. salar organ samples. We found differences in mRNA expression among gills, kidney and intestine. We also found a unique real-time PCR product in S. salar gills through melting curve analysis, agarose gel electrophoresis and cDNA sequencing. This PCR product showed a 98% of identity with the BK channel portion recorded by the NCBI Database and was differentially expressed in gills, kidney and intestine. This real-time PCR assay may become an important tool to study BK potassium channels expressed in the gills of S. salar and its changes during smoltification as putative new candidate to monitor this process.

Producción Científica, Anthocyanins are pigments with antihyperglycemic properties, and they are potential candidates for developing functional foods for the therapy or prevention of Diabetes mellitus type 2 (DM2). The mechanism of these beneficial effects of anthocyanins are, however, hard to explain, given their very low bioavailability due to poor intestinal absorption. We propose that free fatty acid receptor 1 (FFA1, also named GPR40), is involved in an inhibitory effect of the anthocyanidin delphinidin over intestinal glucose absorption. We show the direct effects of delphinidin on the intestine using jejunum samples from RF/J mice, and the human intestinal cell lines HT-29, Caco-2, and NCM460. By the use of specific pharmacological antagonists, we determined that delphinidin inhibits glucose absorption in both mouse jejunum and a human enterocytic cell line in a FFA1-dependent manner. Delphinidin also affects the function of sodium-glucose cotransporter 1 (SGLT1). Intracellular signaling after FFA1 activation involved cAMP increase and cytosolic Ca2+ oscillations originated from intracellular Ca2+ stores and were followed by store-operated Ca2+ entry. Taken together, our results suggest a new GPR-40 mediated local mechanism of action for delphinidin over intestinal cells that may in part explain its antidiabetic effect. These findings are promising for the search for new prevention and pharmacological treatment strategies for DM2 management., INNOVA CORFO (grant 12IDL2-16254), Comisión Nacional de Investigación Científica y Tecnológica (grant 21130644), Ministerio de Economía, Industria y Competitividad (grant BFU2015-70131-R)

Cholinesterase is an enzyme with an unknown physiologic function. Some situations are related to a decreased serum cholinesterase activity but those situations are not well defined and the responsible mechanisms are unknown. Malignancy is one of these situations. The aim of the study was to compare the cholinesterase activity between patients with and without colorectal cancer.Case-control study of patients with colorectal cancer who underwent surgery in our hospital from March 05 to January 07.Patients from the cancer group had a significant decreased serum cholinesterase activity.Investigation of cholinesterase function in humans may result in new research lines in oncology treatment.

In this chapter, a detailed protocol is given for ion-channel reconstitution in the two most used model membranes: planar bilayers and liposomes. In the planar bilayer section, methods are described for the expression of ion channels in Xenopus laevis oocytes, the isolation of their membranes, the insertion of ion channels into the bilayer by vesicle fusion, and the recording of single-ion channel current measurements at a constant applied voltage. The reconstitution of bacterial channels in liposomes is also given. It includes the expression and purification of bacterial channels in E. Coli host strain XL1-blue, the insertion of the channels in liposomes, and the recording of their currents by patch clamping.