Cloning of a proopiomelanocortin cDNA from the pituitary gland of the sea bass (Dicentrarchus labrax) and assessment of mRNA expression in different tissues by means of real-time PCR

Varsamos, Stamatis; Bonga, SE Wendelaar; Flik, G.; Quéré, Ronan; Commes, Thérèse

doi:10.1677/joe.0.1760405

Cited by 33 publications

(17 citation statements)

References 31 publications

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“…In sea bass, no difference was found in pituitary pomc mRNA level in SW or FW fish. In accordance with a previous study (Varsamos et al 2003), pituitary pomc mRNA levels are very high (2-3! 10 8 copies/mg RNA) under both water salinity conditions.…”

Section: Discussionsupporting

confidence: 92%

“…Control or invariant internal 'house-keeping' genes were necessary for the global normalization of the quantification by real-time PCR. The candidate control genes were the L17 ribosomal protein (rpL17), a sea bass house-keeping gene validated previously (Varsamos et al 2003) and elongation factor 1a (ef1a). To determine sea bass ef1a-specific primers, PCR was performed on cDNA obtained from sea bass pituitary RNA with degenerate primers designed on the basis of consensus ef1a sequences (GenBank; http://www.ncbi.nlm.nih.gov/ Genbank/index.html).…”

Section: Primer Design For Target and Reference Genesmentioning

confidence: 99%

“…Quantification of prl, gh, sl, pomc, gthI, and gthII gene expression on sea bass cDNAs in pituitary gland, of igfI expression in liver and of rpL17 and ef1a in both pituitary gland and liver, was carried out by means of real-time quantitative PCR using a Light Cycler (Roche) according to Varsamos et al (2003).…”

Section: Quantification Of Target and Reference Gene Expressionmentioning

confidence: 99%

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Pituitary hormone mRNA expression in European sea bass Dicentrarchus labrax in seawater and following acclimation to fresh water

Varsamos¹,

Xuereb²,

Commes³

et al. 2006

Journal of Endocrinology

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The mRNA expression of pituitary prolactin (prl), growth hormone (gh), somatolactin (sl), proopiomelanocortin (pomc), and gonadotropins (gthI and gthII) was quantified by real-time PCR, in sea bass, Dicentrarchus labrax, adapted for 1 month to seawater (SW) or freshwater (FW). In addition, IGF-I (igfI) mRNA expression in liver and branchial Na C /K C -ATPase activity were determined. L17 ribosomal protein (rpL17) and elongation factor 1a (ef1a) were validated as reference genes in real-time PCR in the experimental context. The real-time PCR assays were validated for the different hormone genes considered. Expression of pituitary pomc, gthI, gthII, gh, and liver igfI was not significantly different between FW and SW fish. Pituitary prl was 4 . 5-fold higher in FW than in SW, whereas pituitary sl was 1 . 8-fold higher in SW-compared with FW-adapted fish. Gill Na C /K C -ATPase specific activity was 2 . 3-fold higher in FW sea bass compared with SW fish. Plasma cortisol levels were 6 . 5-fold lower in SW-than in FW-adapted specimens. The results are discussed in relation to the osmoregulatory strategy of this euryhaline SW species, which displays features that do not fit present models based on salmonids and FW euryhaline teleosts.

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Section: Discussionsupporting

confidence: 92%

Section: Primer Design For Target and Reference Genesmentioning

confidence: 99%

Section: Quantification Of Target and Reference Gene Expressionmentioning

confidence: 99%

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Pituitary hormone mRNA expression in European sea bass Dicentrarchus labrax in seawater and following acclimation to fresh water

Varsamos¹,

Xuereb²,

Commes³

et al. 2006

Journal of Endocrinology

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“…The high levels of POMC transcripts found in the pituitary are consistent with other reports of POMC expression in teleosts (Arends et al 1998, GonzalezNunez et al 2003, Varsamos et al 2003, Karsi et al 2004, Takahashi et al 2005. In rainbow trout, POMC transcripts were detected in most of the other tissues examined, but the levels were low (<40 copies per 0·02 µg RNA) (Fig.…”

Section: Figuresupporting

confidence: 91%

The pro-opiomelanocortin genes in rainbow trout (Oncorhynchus mykiss): duplications, splice variants, and differential expression

Leder¹,

Silverstein²

2006

Journal of Endocrinology

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Pro-opiomelanocortin (POMC) is a precursor for several important peptide hormones involved in a variety of functions ranging from stress response to energy homeostasis. In mammals and fish, the POMC-derived peptide -melanocyte stimulating hormone (MSH) is known to be involved in appetite suppression through its interaction with melanocortin-4 receptors. The details of energy homeostasis in fishes are beginning to be elucidated and many of the genes involved in mammalian neuroendocrine signaling pathways are being discovered in fish. In salmonid fishes such as the rainbow trout, genome duplication adds another degree of complexity when trying to compare gene function and homology with other vertebrates. This is true of the POMC gene. Two copies of the POMC gene were previously identified, A and B, presumably resulting from the salmonid duplication. However, while investigating POMC involvement in the feeding response of rainbow trout, a second copy of POMC-A was discovered which is more likely the result of the salmonid duplication and suggests that POMC-B is a duplicate resulting from the earlier teleost duplication prior to tetrapod divergence. The duplicated POMC-A had five deleted amino acids, five inserted amino acids, and 39 amino acid differences from the published POMC-A. In addition to the duplicate POMC-A, a splice variant of the published POMC-A sequence was also identified. Quantitative real-time PCR assays were developed for the different POMC transcripts, and expression was examined in a variety of tissues. Expression of POMC transcripts was highest in the pituitary for all POMC genes, but varied among other tissues for POMC-A1, POMC-A2, POMC-A2s, and POMC-B. POMC-A1 was the only transcript to respond significantly to food deprivation.

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“…Whilst there have been several reports measuring the precise tissue specific copy number of POMC mRNA (20), the transcriptional regulation of each POMC gene needs to be characterized. The POMC gene is expressed in both the pars distalis (PD) and pars intermedia (PI) of the pituitary gland.…”

Section: Expression Of Pomc Mrnasmentioning

confidence: 99%

Molecular cloning, tissue distribution and quantitative analysis of two proopiomelanocortin mRNAs in Japanese flounder (Paralichthys olivaceus)

Kim

Kim²,

Chen³

et al. 2009

BMB Reports

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Cloning of a proopiomelanocortin cDNA from the pituitary gland of the sea bass (Dicentrarchus labrax) and assessment of mRNA expression in different tissues by means of real-time PCR

Abstract: Proopiomelanocortin (POMC) cDNA was cloned from sea bass (Dicentrarchus labrax) pituitary gland. A 743 nucleotide sequence was obtained coding for the following sequences flanked by sets of proteolytic cleavage sites: ACTH (Ser 88

Cited by 33 publications

References 31 publications

Pituitary hormone mRNA expression in European sea bass Dicentrarchus labrax in seawater and following acclimation to fresh water

Pituitary hormone mRNA expression in European sea bass Dicentrarchus labrax in seawater and following acclimation to fresh water

The pro-opiomelanocortin genes in rainbow trout (Oncorhynchus mykiss): duplications, splice variants, and differential expression

Molecular cloning, tissue distribution and quantitative analysis of two proopiomelanocortin mRNAs in Japanese flounder (Paralichthys olivaceus)

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