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The aim of this study was to investigate the circadian rhythm of muscle-related gene expression in mackerel tuna under different weather conditions. The experiment was carried out under two weather conditions at four sampling times (6:00, 12:00, 18:00, and 24:00) to determine the expression of growth, function, and rhythm genes: white muscle rhythm genes were rhythmic on sunny and cloudy days, except for PER3 and RORA; all functional genes had daily rhythmicity. Red muscle had daily rhythmicity on both sunny and cloudy days; functional genes had daily rhythmicity except for MBNL. The expression levels of the rhythm gene PER1 were determined to be significantly different by independent t-test samples in white muscle at 6:00, 12:00, 18:00, and 24:00 under different weather conditions; the expression levels of the functional genes MBNL and MSTN were both significantly different. In the red muscle, the expression of the rhythm genes PER3, REVERBA, and BMAL1 was determined by independent t-test samples at 6:00, 12:00, 18:00, and 24:00 on cloudy and sunny days; the functional gene MBNL was significantly different. The present study showed that mackerel tuna muscle rhythm genes and functional genes varied significantly in expression levels depending on weather, time of day, and light intensity and that the expression levels of myogenic genes were closely related to clock gene expression. The fish were also able to adapt to changes in light intensity in different weather conditions through positive physiological regulation.

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KEY MESSAGE: Our findings suggest most wheat biological processes are under the control of the daily expressed genes. Plant circadian rhythms represent daily changes in the activity of various processes, which are based on changes in the levels of gene expression and protein synthesis. In wheat, some key components of plant circadian clock have been identified, but there is little data on the daily expression and interactions of these genes. To study the common wheat daily transcriptome, RNA sequencing was performed. Using these data, genes expressed in daily pattern and the metabolic pathways controlled by them were identified: responses to stimuli and nutrients, transport, photoperiodism, photomorphogenesis, synthesis and degradation of different metabolites, and regulation of the processes of RNA synthesis. It was shown that a significant part of the transcriptome can vary greatly daily. Five expression patterns were identified. They were characterized by peaks at different time points and described the genes underlying these patterns. The analysis of the enrichment of gene ontology terms with various patterns allowed us to describe the main metabolic pathways in each group. Wheat homologs of the genes related to circadian clock in Arabidopsis were identified. Most of them were represented by three homoeologous genes expressed uniformly. Comparison of their expression patterns demonstrated a shift in the expression peaks for some core and accessory genes; the majority of wheat circadian genes were expressed in accordance with Arabidopsis homologs. This may indicate a similar functional role of these genes in wheat.

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The hormone melatonin connects environmental cues, such as photoperiod and temperature, with a number of physiological and behavioural processes, including seasonal reproduction, through binding to their cognate receptors. This study reports the structural, functional and physiological characterization of five high-affinity melatonin receptors (Mtnr1aaα, Mtnr1aaß, Mtnr1ab, Mtnr1al, Mtnr1b) in Atlantic salmon. Phylogenetic analysis clustered salmon melatonin receptors into three monophyletic groups, Mtnr1A, Mtnr1Al and Mtnr1B, but no functional representative of the Mtnr1C group. Contrary to previous studies in vertebrates, pharmacological characterization of four receptors in COS-7, CHO and SH-SY5Y cell lines (Mtnr1Aaα, Mtnr1Aaß, Mtnr1Ab, Mtnr1B) showed induction of intracellular cAMP levels following 2-iodomelatonin or melatonin exposure. No consistent response was measured after N-acetyl-serotonin or serotonin exposure. Melatonin receptor genes were expressed at all levels of the hypothalamo-pituitary-gonad axis, with three genes (mtnr1aaß, mtnr1ab and mtnr1b) detected in the pituitary. Pituitary receptors displayed daily fluctuations in mRNA levels during spring, prior to the onset of gonadal maturation, but not in autumn, strongly implying a direct involvement of melatonin in seasonal processes regulated by the pituitary. To the best of our knowledge, this is the first report of cAMP induction mediated via melatonin receptors in a teleost species.

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This research aimed at investigating circadian rhythm expression of key genes involved in lipid metabolism in the liver of a teleost fish (Sparus aurata), and their synchronisation to different light-dark (L-D) and feeding cycles. To this end, 90 gilthead sea bream were kept in 12:12 h (light:dark, LD, lights on at ZT0) and fed a single daily meal at mid-light (ML = ZT6), mid-darkness (MD = ZT18) and randomly (RD) at a 1.5% body weight ration. A total of 18 tanks were used, six tanks per feeding treatment with five fishes per tank; locomotor activity was recorded in each tank. After 25 days of synchronisation to these feeding regimes, fishes were fasted for one day and liver samples were taken every 4 hours during a 24 h cycle (ZT2, 6, 10, 14, 18 and 22) and stored at -80 °C until analysis. To determine whether the rhythm expression presented an endogenous control, another experiment was performed using 30 fish kept in complete darkness and fed randomly (DD/RD). Samples were taken following the same procedure as above. The results revealed that all genes investigated exhibited well defined daily rhythms. The lipolysis-related and fatty acid turnover genes (hormone-sensitive lipase (hsl) and peroxisome proliferator-activated receptor-α (pparα)) exhibited a nocturnal achrophase (Ø = ZT18:03-19:21); lipoprotein lipase (lpl) also showed the same nocturnal achrophase (Ø = ZT20:04-21:36). In contrast, lipogenesis-related gene, fatty acid synthase (fas), and of fatty acid turnover, cyclooxygenase (cox-2), showed a diurnal rhythm (Ø = ZT2:27-8:09); while pparγ was nocturnal (Ø = ZT16:16-18:05). Curiously, feeding time had little influence on the phase of these daily rhythms, since all feeding groups displayed similar achrophases. Furthermore, under constant conditions pparα and hsl showed circadian rhythmicity. These findings suggest that lipid utilisation in the liver is rhythmic and strongly synchronised to the LD cycle, regardless of feeding time, which should be taken into consideration when investigating fish nutrition and the design of feeding protocols.

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