Abstract

BackgroundNesfatin-1 is a recently discovered anorexigen encoded in the precursor peptide, nucleobindin-2 (NUCB2) in mammals. To date, nesfatin-1 has not been described in any non-mammalian species, although some information is available in the sequenced genomes of several species. Our objective was to characterize nesfatin-1 in fish.Methodology/Principal FindingsIn the present study, we employed molecular, immunohistochemical, and physiological studies to characterize the structure, distribution, and appetite regulatory effects of nesfatin-1 in a non-mammalian vertebrate. A very high conservation in NUCB2 sequences, especially in the nesfatin-1 region was found in lower vertebrates. Abundant expression of NUCB2 mRNA was detected in several tissues including the brain and liver of goldfish. Nesfatin-1-like immunoreactive cells are present in the feeding regulatory nucleus of the hypothalamus and in the gastrointestinal tract of goldfish. Approximately 6-fold increase in NUCB2 mRNA levels was found in the liver after 7-day food-deprivation, and a similar increase was also found after short-term fasting. This points toward a possible liver specific role for NUCB2 in the control of metabolism during food-deprivation. Meanwhile, ∼2-fold increase at 1 and 3 h post-feeding and an ∼3-fold reduction after a 7-day food-deprivation was observed in NUCB2 mRNA in the goldfish hypothalamus. In vivo, a single intraperitoneal injection of the full-length native (goldfish; gf) nesfatin-1 at a dose of 50 ng/g body weight induced a 23% reduction of food intake one hour post-injection in goldfish. Furthermore, intracerebroventricular injection of gfnesfatin-1 at a dose of 5 ng/g body weight resulted in ∼50% reduction in food intake.Conclusions/SignificanceOur results provide molecular, anatomical and functional evidences to support potential anorectic and metabolic roles for endogenous nesfatin-1 in goldfish. Collectively, we provide novel information on NUCB2 in non-mammals and an anorexigenic role for nesfatin-1 in goldfish.

Highlights

  • Nesfatin-1 is a recently identified anorectic peptide cleaved from the N-terminal of the precursor protein nucleobindin 2 (NUCB2) [1]

  • NUCB2A in Takifugu rubripes and Tetraodon nigroviridis both have 12 exons interspaced by 11 introns and this appears to be due to the deletion of exon 1. (Figure 1A)

  • There have been no reports on the processing of nesfatin-1 from NUCB1 or any reports of the involvement of NUCB1 in food intake or glucose homeostasis

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Summary

Introduction

Nesfatin-1 is a recently identified anorectic peptide cleaved from the N-terminal of the precursor protein nucleobindin 2 (NUCB2) [1]. Nesfatin-1 has been detected in both rat cerebrospinal fluid [1] and in the peripheral circulation of humans and rodents [9,12,13,14]. Acute administration of nesfatin-1, by central or peripheral injections has been shown to reduce food intake in rats for up to 6 hours [1,13,14,15]. Nesfatin-1 has been shown to regulate cardiac functions [16], reduce blood glucose levels in diabetic mice [14], and induce fear and anxiety like behaviors [17]. Nesfatin-1 is a recently discovered anorexigen encoded in the precursor peptide, nucleobindin-2 (NUCB2) in mammals.

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