Acidic Digestion in a Teleost: Postprandial and Circadian Pattern of Gastric pH, Pepsin Activity, and Pepsinogen and Proton Pump mRNAs Expression

Two different modes for regulation of stomach acid secretion have been described in vertebrates. Some species exhibit a continuous acid secretion maintaining a low gastric pH during fasting. Others, as some teleosts, maintain a neutral gastric pH during fasting while the hydrochloric acid is released only after the ingestion of a meal. Those different patterns seem to be closely related to specific feeding habits. However, our recent observations suggest that this acidification pattern could be modified by changes in daily feeding frequency and time schedule. The aim of this study was to advance in understanding the regulation mechanisms of stomach digestion and pattern of acid secretion in teleost fish. We have examined the postprandial pattern of gastric pH, pepsin activity, and mRNA expression for pepsinogen and proton pump in white seabream juveniles maintained under a light/dark 12/12 hours cycle and receiving only one morning meal. The pepsin activity was analyzed according to the standard protocol buffering at pH 2 and using the actual pH measured in the stomach. The results show how the enzyme precursor is permanently available while the hydrochloric acid, which activates the zymogen fraction, is secreted just after the ingestion of food. Results also reveal that analytical protocol at pH 2 notably overestimates true pepsin activity in fish stomach. The expression of the mRNA encoding pepsinogen and proton pump exhibited almost parallel patterns, with notable increases during the darkness period and sharp decreases just before the morning meal. These results indicate that white seabream uses the resting hours for recovering the mRNA stock that will be quickly used during the feeding process. Our data clearly shows that both daily illumination pattern and feeding time are involved at different level in the regulation of the secretion of digestive juices

Defining Global Gene Expression Changes of the Hypothalamic-Pituitary-Gonadal Axis in Female sGnRH-Antisense Transgenic Common Carp (Cyprinus carpio)

BACKGROUND: The hypothalamic-pituitary-gonadal (HPG) axis is critical in the development and regulation of reproduction in fish. The inhibition of neuropeptide gonadotropin-releasing hormone (GnRH) expression may diminish or severely hamper gonadal development due to it being the key regulator of the axis, and then provide a model for the comprehensive study of the expression patterns of genes with respect to the fish reproductive system. METHODOLOGY/PRINCIPAL FINDINGS: In a previous study we injected 342 fertilized eggs from the common carp (Cyprinus carpio) with a gene construct that expressed antisense sGnRH. Four years later, we found a total of 38 transgenic fish with abnormal or missing gonads. From this group we selected the 12 sterile females with abnormal ovaries in which we combined suppression subtractive hybridization (SSH) and cDNA microarray analysis to define changes in gene expression of the HPG axis in the present study. As a result, nine, 28, and 212 genes were separately identified as being differentially expressed in hypothalamus, pituitary, and ovary, of which 87 genes were novel. The number of down- and up-regulated genes was five and four (hypothalamus), 16 and 12 (pituitary), 119 and 93 (ovary), respectively. Functional analyses showed that these genes involved in several biological processes, such as biosynthesis, organogenesis, metabolism pathways, immune systems, transport links, and apoptosis. Within these categories, significant genes for neuropeptides, gonadotropins, metabolic, oogenesis and inflammatory factors were identified. CONCLUSIONS/SIGNIFICANCE: This study indicated the progressive scaling-up effect of hypothalamic sGnRH antisense on the pituitary and ovary receptors of female carp and provided comprehensive data with respect to global changes in gene expression throughout the HPG signaling pathway, contributing towards improving our understanding of the molecular mechanisms and regulative pathways in the reproductive system of teleost fish

Vitamin C Enhances Vitamin E Status and Reduces Oxidative Stress Indicators in Sea Bass Larvae Fed High DHA Microdiets

Docosahexaenoic acid (DHA) is an essential fatty acid necessary for many biochemical, cellular and physiological functions in fish. However, high dietary levels of DHA increase free radical injury in sea bass (Dicentrarchus labrax) larvae muscle, even when vitamin E (α-tocopherol, α-TOH) is increased. Therefore, the inclusion of other nutrients with complementary antioxidant functions, such as vitamin C (ascorbic acid, vitC), could further contribute to prevent these lesions. The objective of the present study was to determine the effect of vitC inclusion (3,600 mg/kg) in high DHA (5 % DW) and α-TOH (3,000 mg/kg) microdiets (diets 5/3,000 and 5/3,000 + vitC) in comparison to a control diet (1 % DHA DW and 1,500 mg/kg of α-TOH; diet 1/1,500) on sea bass larvae growth, survival, whole body biochemical composition and thiobarbituric acid reactive substances (TBARS) content, muscle morphology, skeletal deformities and antioxidant enzymes, insulin-like growth factors (IGFs) and myosin expression (MyHC). Larvae fed diet 1/1,500 showed the best performance in terms of total length, incidence of muscular lesions and ossification degree. IGFs gene expression was elevated in 5/3,000 diet larvae, suggesting an increased muscle mitogenesis that was confirmed by the increase in the mRNA copies of MyHC. vitC effectively controlled oxidative damages in muscle, increased α-TOH larval contents and reduced TBARS content and the occurrence of skull deformities. The results of the present study showed the antioxidant synergism between vitamins E and C when high contents of DHA are included in sea bass larvae diets

Transient up- and down-regulation of expression of myosin light chain 2 and myostatin mRNA mark the changes from stratified hyperplasia to muscle fiber hypertrophy in larvae of gilthead sea bream (Sparus aurata L.)

Hyperplasia and hypertrophy are the two mechanisms by which muscle develops and grows. We study these two mechanisms, during the early development of white muscle in Sparus aurata, by means of histology and the expression of structural and regulatory genes. A clear stage of stratified hyperplasia was identified early in the development of gilthead sea bream but ceased by 35 dph when hypertrophy took over. Mosaic recruitment of new white fibers began as soon as 60 dph. The genes mlc2a and mlc2b were expressed at various levels during the main phases of hyperplasia and hypertrophy. The genes myog and mlc2a were significantly up-regulated during the intensive stratified formation of new fibers and their expression was significantly correlated. Expression of mstn1 and igf1 increased at 35 dph, appeared to regulate the hyperplasia-to-hypertrophy transition, and may have stimulated the expression of mlc2a, mlc2b and col1a1 at the onset of mosaic hyperplasia. The up-regulation of mstn1 at transitional phases in muscle development indicates a dual regulatory role of myostatin in fish larval muscle growth

LOFAR 144-MHz follow-up observations of GW170817

ABSTRACT We present low-radio-frequency follow-up observations of AT 2017gfo, the electromagnetic counterpart of GW170817, which was the first binary neutron star merger to be detected by Advanced LIGO–Virgo. These data, with a central frequency of 144 MHz, were obtained with LOFAR, the Low-Frequency Array. The maximum elevation of the target is just 13${_{.}^{\circ}}$7 when observed with LOFAR, making our observations particularly challenging to calibrate and significantly limiting the achievable sensitivity. On time-scales of 130–138 and 371–374 d after the merger event, we obtain 3σ upper limits for the afterglow component of 6.6 and 19.5 mJy beam−1, respectively. Using our best upper limit and previously published, contemporaneous higher frequency radio data, we place a limit on any potential steepening of the radio spectrum between 610 and 144 MHz: the two-point spectral index $\alpha ^{610}_{144} \gtrsim$ −2.5. We also show that LOFAR can detect the afterglows of future binary neutron star merger events occurring at more favourable elevations.</jats:p

Effects of retinoic acid (RA) on gene expression of nuclear receptors and genes involved in the synthesis and degradation in Senegalese sole larvae (Solea senegalensis)

Retinoic acid (RA) is a potent morphogenetic nutrient that plays an essential role in the processes of morphogenesis, skeletogenesis and pigmentation of fishes by activating the expression of over 500 genes. However, still little is known about the transcriptional cascades mediated by RA in flatfish species. Therefore, the aim of this study was to evaluate the effects of RA during the larval development and metamorphosis of Senegalese sole and to determine the transcriptional responses of genes involved in RA metabolism and action. In a first experiment, pre-metamorphic larvae (7 days after hatching, dah) were treated with an inhibitor of RA synthesis (DEAB) until the end of metamorphosis (20dah). In a second experiment, post-metamorphic larvae pretreated for 3 days with DEAB and DMSO (solvent of DEAB) were subdivided into three additional tanks to which all-trans RA (ATRA) and TTNPB (rar agonist) were added. In each experiment, samples were taken at different times to quantify the expression of genes involved in RA synthesis (rdh10a, raldh2), degradation (cyP26a1), metabolism (rbp1, rbp4, crabp1a), mediators of its action (retinoic acid nuclear receptors, rar&alpha;1, rar&alpha;2, rar&gamma;##, rxr&alpha;, rxr&beta;1, rxr&beta;2, ror&beta;, rarres3) and other RA-related pathways (thr&alpha;a, thr&alpha;b, thr&beta;). &nbsp;Retinoic acid (RA) is a potent morphogenetic nutrient that plays an essential role in the processes of morphogenesis, skeletogenesis and pigmentation of fishes by activating the expression of over 500 genes. However, still little is known about the transcriptional cascades mediated by RA in flatfish species. Therefore, the aim of this study was to evaluate the effects of RA during the larval development and metamorphosis of Senegalese sole and to determine the transcriptional responses of genes involved in RA metabolism and action. In a first experiment, pre-metamorphic larvae (7 days after hatching, dah) were treated with an inhibitor of RA synthesis (DEAB) until the end of metamorphosis (20dah). In a second experiment, post-metamorphic larvae pretreated for 3 days with DEAB and DMSO (solvent of DEAB) were subdivided into three additional tanks to which all-trans RA (ATRA) and TTNPB (rar agonist) were added. In each experiment, samples were taken at different times to quantify the expression of genes involved in RA synthesis (rdh10a, raldh2), degradation (cyP26a1), metabolism (rbp1, rbp4, crabp1a), mediators of its action (retinoic acid nuclear receptors, rar&alpha;1, rar&alpha;2, rar&gamma;##, rxr&alpha;, rxr&beta;1, rxr&beta;2, ror&beta;, rarres3) and other RA-related pathways (thr&alpha;a, thr&alpha;b, thr&beta;). &nbsp;</p