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      Effect of Fasting on the Spexin System in Broiler Chickens

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          Abstract

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          The regulation of physiological processes by biologically active substances such as peptides, proteins, or hormones is very important in the context of both the development of the basic sciences and their subsequent use in improving animal husbandry. One such substance is spexin (SPX), a recently discovered, very conservative peptide that has been shown in mammalian studies to be able to regulate food intake, as well as carbohydrate–lipid metabolism. Because there is no information on the role of SPX in the metabolism of birds in the literature, we first decided to determine whether the expression of the gene encoding this peptide is present in the various tissues of birds. A further object of the study was to determine whether the concentration of SPX in serum blood changes during the disturbance of the carbohydrate metabolism caused by starvation and whether these changes correlate with other metabolic parameters. These studies will help us fully understand the role of SPX in bird physiology, and this research should be further extended.

          Abstract

          Spexin (SPX) is a highly conservative peptide hormone containing 14 amino acids and was discovered in 2007 by bioinformatics methods. However, nothing is yet known about its role in the metabolism of birds, including broilers. The aim of this study was to investigate the effect of short-term fasting (2, 4, and 8 h) on the concentration of SPX in blood serum and the expression levels of the genes encoding this peptide ( SPX1) and its receptors, GALR2 and GALR3, in the tissues involved in carbohydrate and lipid metabolism (muscles, adipose tissue, and liver). We also analyzed the mRNA expression of these genes in various chicken tissues. Moreover, we studied the correlation between the serum level of SPX and other metabolic parameters (insulin, glucagon, glucose, triglycerides, and cholesterol). Using RT-qPCR, we found that SPX1, GALR2, and GALR3 are expressed in all investigated tissues in broiler chicken. Moreover, using a commercially available radio-immunoassay, we noted an increase of the SPX level in blood serum after 4 and 8 h of fasting compared to nonfasted animals ( p < 0.05). This increase was positively correlated with glucagon concentration (r = 0.341; p < 0.05) and negatively with glucose concentration (r = −0.484; p < 0.01). Additionally, we discovered that in the short term, food deprivation leads to the expression regulation of SPX1, GALR2, and GLAR3 in tissues associated with metabolism of carbohydrates and lipids. The obtained results indicate that SPX is involved in the regulation of metabolism in broiler chickens.

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          Most cited references27

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          Identification of novel peptide hormones in the human proteome by hidden Markov model screening.

          Peptide hormones are small, processed, and secreted peptides that signal via membrane receptors and play critical roles in normal and pathological physiology. The search for novel peptide hormones has been hampered by their small size, low or restricted expression, and lack of sequence similarity. To overcome these difficulties, we developed a bioinformatics search tool based on the hidden Markov model formalism that uses several peptide hormone sequence features to estimate the likelihood that a protein contains a processed and secreted peptide of this class. Application of this tool to an alignment of mammalian proteomes ranked 90% of known peptide hormones among the top 300 proteins. An analysis of the top scoring hypothetical and poorly annotated human proteins identified two novel candidate peptide hormones. Biochemical analysis of the two candidates, which we called spexin and augurin, showed that both were localized to secretory granules in a transfected pancreatic cell line and were recovered from the cell supernatant. Spexin was expressed in the submucosal layer of the mouse esophagus and stomach, and a predicted peptide from the spexin precursor induced muscle contraction in a rat stomach explant assay. Augurin was specifically expressed in mouse endocrine tissues, including pituitary and adrenal gland, choroid plexus, and the atrio-ventricular node of the heart. Our findings demonstrate the utility of a bioinformatics approach to identify novel biologically active peptides. Peptide hormones and their receptors are important diagnostic and therapeutic targets, and our results suggest that spexin and augurin are novel peptide hormones likely to be involved in physiological homeostasis.
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            Coevolution of the spexin/galanin/kisspeptin family: Spexin activates galanin receptor type II and III.

            The novel neuropeptide spexin (SPX) was discovered using bioinformatics. The function of this peptide is currently under investigation. Here, we identified SPX along with a second SPX gene (SPX2) in vertebrate genomes. Syntenic analysis and relocating SPXs and their neighbor genes on reconstructed vertebrate ancestral chromosomes revealed that SPXs reside in the near vicinity of the kisspeptin (KISS) and galanin (GAL) family genes on the chromosomes. Alignment of mature peptide sequences showed some extent of sequence similarity among the 3 peptide groups. Gene structure analysis indicated that SPX is more closely related to GAL than KISS. These results suggest that the SPX, GAL, and KISS genes arose through local duplications before 2 rounds (2R) of whole-genome duplication. Receptors of KISS and GAL (GAL receptor [GALR]) are phylogenetically closest among rhodopsin-like G protein-coupled receptors, and synteny revealed the presence of 3 distinct receptor families KISS receptor, GALR1, and GALR2/3 before 2R. A ligand-receptor interaction study showed that SPXs activate human, Xenopus, and zebrafish GALR2/3 family receptors but not GALR1, suggesting that SPXs are natural ligands for GALR2/3. Particularly, SPXs exhibited much higher potency toward GALR3 than GAL. Together, these results identify the coevolution of SPX/GAL/KISS ligand genes with their receptor genes. This study demonstrates the advantage of evolutionary genomics to explore the evolutionary relationship of a peptide gene family that arose before 2R by local duplications.
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              Spexin is a Novel Human Peptide that Reduces Adipocyte Uptake of Long Chain Fatty Acids and Causes Weight Loss in Rodents with Diet-induced Obesity*

              Objective Microarray studies identified Ch12:orf39 (Spexin) as the most dysregulated gene in obese human fat. Therefore we examined its role in obesity pathogenesis. Design and Methods Spexin effects on food intake, meal patterns, body weight, Respiratory Exchange Ratio (RER), and locomotor activity were monitored electronically in C57BL/6J mice or Wistar rats with dietary-induced obesity (DIO). Its effects on adipocyte [3H]-oleate uptake were determined. Results In humans, Spexin gene expression was down-regulated 14.9-fold in obese omental and subcutaneous fat. Circulating Spexin changed in parallel, correlating (r = −0.797) with Leptin. In rats, Spexin (35 μg/kg/day s.c) reduced caloric intake ~32% with corresponding weight loss. Meal patterns were unaffected. In mice, Spexin (25 μg/kg/day i.p.) significantly reduced the RER at night, and increased locomotion. Spexin incubation in vitro significantly inhibited facilitated fatty acid (FA) uptake into DIO mouse adipocytes. Conditioned taste aversion testing (70μg/kg/day i.p.) demonstrated no aversive Spexin effects. Conclusions Spexin gene expression is markedly down-regulated in obese human fat. The peptide produces weight loss in DIO rodents. Its effects on appetite and energy regulation are presumably central; those on adipocyte FA uptake appear direct and peripheral. Spexin is a novel hormone involved in weight regulation, with potential for obesity therapy.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Animals (Basel)
                Animals (Basel)
                animals
                Animals : an Open Access Journal from MDPI
                MDPI
                2076-2615
                17 February 2021
                February 2021
                : 11
                : 2
                : 518
                Affiliations
                [1 ]Department of Animal Physiology, Biochemistry and Biostructure, Poznan University of Life Sciences, 60-637 Poznan, Poland; ewa.pruszynska@ 123456up.poznan.pl (E.P.-O.); maciej.sassek@ 123456up.poznan.pl (M.S.); natalia.leciejewska@ 123456up.poznan.pl (N.L.); jakub.bien@ 123456up.poznan.pl (J.B.)
                [2 ]Department of Animal Breeding and Product Quality Assessment, Poznan University of Life Sciences, Sloneczna 1, 62-002 Zlotniki, Poland; marcin.hejdysz@ 123456up.poznan.pl (M.H.); piotr.slosarz@ 123456up.poznan.pl (P.Ś.)
                [3 ]Laboratory of Neurobiology, Department of Zoology, Faculty of Veterinary Medicine and Animal Science, Poznan University of Life Sciences, 60-625 Poznan, Poland; kamil.ziarniak@ 123456up.poznan.pl
                [4 ]Department of Preclinical Sciences and Infectious Diseases, Faculty of Veterinary Medicine and Animal Science, Poznan University of Life Sciences, 60-637 Poznan, Poland
                [5 ]Department of Animal Nutrition, Poznan University of Life Sciences, 60-637 Poznan, Poland; marta.kubis@ 123456up.poznan.pl (M.K.); sebastian.kaczmarek@ 123456up.poznan.pl (S.K.)
                Author notes
                [* ]Correspondence: pawel.kolodziejski@ 123456up.poznan.pl or pawelbigi@ 123456o2.pl ; Tel.: +48-511-468-396
                Author information
                https://orcid.org/0000-0003-0715-0223
                https://orcid.org/0000-0003-1772-7997
                https://orcid.org/0000-0003-3179-503X
                https://orcid.org/0000-0003-2732-2344
                https://orcid.org/0000-0003-3638-4192
                Article
                animals-11-00518
                10.3390/ani11020518
                7922423
                33671411
                9c31c95e-49c4-4c02-8a91-6bdd0d1c2f66
                © 2021 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 16 January 2021
                : 15 February 2021
                Categories
                Article

                spexin,broiler chicken,fasting
                spexin, broiler chicken, fasting

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