Ghrelin O-Acyl Transferase in Zebrafish Is an Evolutionarily Conserved Peptide Upregulated During Calorie Restriction

Hatef, Azadeh; Yufa, Roman; Unniappan, Suraj

doi:10.1089/zeb.2014.1062

Cited by 26 publications

(20 citation statements)

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“…Food deprivation is known to play a significant role in altering neuroendocrine factors that play critical role in reproduction and energy intake in several species 13 24 . In goldfish, fasting for 7 or 28 days did not result in any significant changes in body weight and this result is in agreement with previous reports in goldfish 25 26 27 . NUCB1 expression in gut was significantly lower when fish were food deprived for one week 13 .…”

Section: Discussionsupporting

confidence: 93%

“…For IHC studies conducted as described in detail earlier1626, pituitary, ovary, J-loop and testis were collected from goldfish. The antibodies used were: rabbit polyclonal anti- mouse nucleobindin-1 (Pacific immunology, Ramona, CA) for NUCB1, mouse monoclonal anti-growth hormone (Catalog # CLX 130AP, 1:500, Cedarlane, USA) for growth hormone in pituitary, mouse monoclonal anti-ghrelin hormone (Catalog # ab57222, 1:500, Abcam, Massachusetts) for ghrelin in J-Loop11, mouse monoclonal anti-SOX9 hormone (Catalog # ab76997, 1:500, Abcam, Massachusetts) for SOX9 in testis2033, goat monoclonal anti-FOXL2 hormone (Catalog # PA5-18175, 1:500, Waltham, MA ) for FOXL22033 staining in ovary respectively.…”

Section: Methodsmentioning

confidence: 99%

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Nesfatin-1-Like Peptide Encoded in Nucleobindin-1 in Goldfish is a Novel Anorexigen Modulated by Sex Steroids, Macronutrients and Daily Rhythm

Sundarrajan

Blanco

Bertucci

et al. 2016

Sci Rep

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Nesfatin-1 is an 82 amino acid anorexigen encoded in a secreted precursor nucleobindin-2 (NUCB2). NUCB2 was named so due to its high sequence similarity with nucleobindin-1 (NUCB1). It was recently reported that NUCB1 encodes an insulinotropic nesfatin-1-like peptide (NLP) in mice. Here, we aimed to characterize NLP in fish. RT- qPCR showed NUCB1 expression in both central and peripheral tissues. Western blot analysis and/or fluorescence immunohistochemistry determined NUCB1/NLP in the brain, pituitary, testis, ovary and gut of goldfish. NUCB1 mRNA expression in goldfish pituitary and gut displayed a daily rhythmic pattern of expression. Pituitary NUCB1 mRNA expression was downregulated by estradiol, while testosterone upregulated its expression in female goldfish brain. High carbohydrate and fat suppressed NUCB1 mRNA expression in the brain and gut. Intraperitoneal injection of synthetic rat NLP and goldfish NLP at 10 and 100 ng/g body weight doses caused potent inhibition of food intake in goldfish. NLP injection also downregulated the expression of mRNAs encoding orexigens, preproghrelin and orexin-A, and upregulated anorexigen cocaine and amphetamine regulated transcript mRNA in goldfish brain. Collectively, these results provide the first set of results supporting the anorectic action of NLP, and the regulation of tissue specific expression of goldfish NUCB1.

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Section: Discussionsupporting

confidence: 93%

Section: Methodsmentioning

confidence: 99%

Nesfatin-1-Like Peptide Encoded in Nucleobindin-1 in Goldfish is a Novel Anorexigen Modulated by Sex Steroids, Macronutrients and Daily Rhythm

Sundarrajan

Blanco

Bertucci

et al. 2016

Sci Rep

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“…This indicates that GOAT might be restrictedly distributed within the mammalian brain and then, its transcripts are detected when discrete locations are analysed separately. In fish, the only study available points out that GOAT seems to be more widely distributed within the brain, as important levels of goat mRNA expression are observed in the whole brain of zebrafish using RT‐qPCR (Hatef et al, ). Results from the present study show a broad expression of goat in the goldfish brain by RT‐qPCR, in agreement with zebrafish.…”

Section: Discussionmentioning

confidence: 99%

“…Although in lower levels, mRNAs encoding this enzyme were also detected in hypothalamus, pituitary, taste cells, heart, kidney, ovary, placenta, adipose tissue, muscle, serum and chondrocytes of mammals (Gonz alez et al, 2008;G omez et al, 2009;Sakata et al, 2009;Gahete et al, 2010;O'Brien et al, 2010;Shin et al, 2010;Rucinski et al, 2012). In fish, a recent study has shown that GOAT is mainly expressed in gut, gonads and brain of zebrafish (Hatef et al, 2015). Given that the gastrointestinal tract is the main tissue expressing GOAT, further studies using imaging techniques have demonstrated that GOAT is specifically located in the gastric mucosal cells (Sakata et al, 2009;Stengel et al, 2010;Hatef et al, 2015) where it colocalizes with ghrelin in both mammals (Sakata et al, 2009;Stengel et al, 2010) and fish (Hatef et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Brain Mapping of Ghrelin O‐Acyltransferase in Goldfish (Carassius Auratus): Novel Roles for the Ghrelinergic System in Fish?

Blanco

Sánchez-Bretaño

Delgado

et al. 2016

The Anatomical Record

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Ghrelin O-acyltransferase (GOAT) is the enzyme responsible for acylation of ghrelin, a gut-brain hormone with important roles in many physiological functions in vertebrates. Many aspects of GOAT remain to be elucidated, especially in fish, and particularly its anatomical distribution within the different brain areas has never been reported to date. The present study aimed to characterize the brain mapping of GOAT using RT-qPCR and immunohistochemistry in a teleost, the goldfish (Carassius auratus). Results show that goat transcripts are expressed in different brain areas of the goldfish, with the highest levels in the vagal lobe. Using immunohistochemistry, we also report the presence of GOAT immunoreactive cells in different encephalic areas, including the telencephalon, some hypothalamic nuclei, pineal gland, optic tectum and cerebellum, although they are especially abundant in the hindbrain. Particularly, an important signal is observed in the vagal lobe and some fiber tracts of the brainstem, such as the medial longitudinal fasciculus, Mauthneri fasciculus, secondary gustatory tract and spinothalamic tract. Most of the forebrain areas where GOAT is detected, particularly the hypothalamic nuclei, also express the ghs-r1a ghrelin receptor and other appetite-regulating hormones (e.g., orexin and NPY), supporting the role of ghrelin as a modulator of food intake and energy balance in fish. Present results are the first report on the presence of GOAT in the brain using imaging techniques. The high presence of GOAT in the hindbrain is a novelty, and point to possible new functions for the ghrelinergic system in fish. Anat Rec, 299:748-758, 2016. V C 2016 Wiley Periodicals, Inc.

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“…tive cells in the gonads will assist us in predicting possible reproductive regulatory roles of PNX in zebrafish. Immunohistochemical studies (IHC) were conducted using methods described earlier 35 . The details of antibodies used in our research are rabbit anti PNX-14 primary antibody (Phoenix Pharmaceuticals, Burlingame, CA, United States, 1:1000 dilution) and rabbit anti-SREB3 antibody -N-terminal (Abcam, Cambridge, UK, 1:500 dilution).…”

Section: Immunohistochemial and Immunocytochemical Localization Of Pnmentioning

confidence: 99%

Phoenixin-20 Stimulates mRNAs Encoding Hypothalamo-Pituitary-Gonadal Hormones, is Pro-Vitellogenic, and Promotes Oocyte Maturation in Zebrafish

Rajeswari

Unniappan

2020

Sci Rep

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Phoenixin-20 (PNX-20) is a bioactive peptide with hormone-like actions in vertebrates. In mammals, PNX stimulates hypothalamo-pituitary-gonadal hormones and regulate reproductive processes. Our immunohisto/cytochemical studies show PNX-like and the putative PNX receptor, SREB3-like immunoreactivity in the gonads of zebrafish, and in zebrafish liver (ZFL) cells. Intraperitoneal injection of zebrafish PNX-20 upregulates mRNAs encoding both salmon gonadotropin-releasing hormone (GnRH), and chicken GnRH-II and kisspeptin and its receptor in zebrafish hypothalamus. Similarly, luteinizing hormone receptor mRNA expression in the testis, follicle-stimulating hormone receptor in the ovary, and the kisspeptin system were upregulated in the gonads of PNX-20 injected fish. We also observed the upregulation of genes involved in the sex steroidogenic pathway (cyp11a1, cyp17a1, 17βhsd, cyp19a1a) in the gonads of PNX-20 administered fish. PNX-20 upregulates the expression of vitellogenin isoforms and estrogen receptor (esr2a and 2b) mRNAs in ZFL cells in vitro. Meanwhile, siRNA-mediated knockdown of PNX-20 resulted in the downregulation of all vitellogenin transcripts, further suggesting its possible role in vitellogenesis. PNX-20 treatment resulted in a significant increase in germinal vesicle breakdown in zebrafish follicles in vitro. Collectively, these results provide strong evidence for PNX-20 effects on the HPG axis and liver to promote reproduction in zebrafish. Advances in computational biology, especially in the use of bioinformatics tools, helped in the discovery of many regulatory molecules with hormone-like actions. A recent example is phoenixin-20 (PNX-20), which was originally identified as a reproductive regulatory peptide 1. Phoenixin was first isolated from the rat hypothalamus and bovine heart by two independent research groups 1,2. The mature phoenixin peptide is produced from an uncharacterized protein called small integral membrane protein 20 (SMIM20) or chromosome 4 open reading frame 52 (C4orf52). Phoenixin is reported to be present in two active forms, a 20 amino acid peptide named PNX-20 and a 14 amino acid one called PNX-14 1,2. PNX-14 is identical in humans, rats, mice, pigs, and dogs. PNX-20 differs in one amino acid between the coding regions of human, canine and porcine sequences 1. It was reported that PNX is expressed in various parts of the brain and peripheral tissues in mammals, with the highest expression in the hypothalamus. Treen et al., reported that a putative receptor of PNX is the G protein-coupled receptor 173 (GPR173), which is also called super conserved receptor expressed in brain 3 (SREB3) 3. PNX stimulates reproductive functions via acting on the HPG axis 1. Later it was reported that PNX mediates its reproductive regulatory effects through GPR173, and cAMP-PKA dependent pathway, acting on both gonadotropin-releasing hormone (GnRH) and kisspeptin 1 (Kiss1) expressing neurons 3. In mice, PNX positively influences Kiss1 transcription and GnRH mediated gonadotropin release 1,4. Intrac...

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Ghrelin O-Acyl Transferase in Zebrafish Is an Evolutionarily Conserved Peptide Upregulated During Calorie Restriction

Cited by 26 publications

References 58 publications

Nesfatin-1-Like Peptide Encoded in Nucleobindin-1 in Goldfish is a Novel Anorexigen Modulated by Sex Steroids, Macronutrients and Daily Rhythm

Nesfatin-1-Like Peptide Encoded in Nucleobindin-1 in Goldfish is a Novel Anorexigen Modulated by Sex Steroids, Macronutrients and Daily Rhythm

Brain Mapping of Ghrelin O‐Acyltransferase in Goldfish (Carassius Auratus): Novel Roles for the Ghrelinergic System in Fish?

Phoenixin-20 Stimulates mRNAs Encoding Hypothalamo-Pituitary-Gonadal Hormones, is Pro-Vitellogenic, and Promotes Oocyte Maturation in Zebrafish

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