Unknown,Transcriptomics,Genomics,Proteomics

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Skin healing and scale regeneration in fed and unfed sea bream, Sparus auratus

ABSTRACT: Fish scales are an important reservoir of calcium and phosphorus and together with the skin function as an integrated barrier against environmental changes and external aggressors. Histological studies have revealed that the skin and scales regenerate rapidly in fish when they are lost or damaged. In the present manuscript the histological and molecular changes underlying skin and scale regeneration in fed and fasted sea bream (Sparus auratus) were studied using a microarray 3 and 7 days after scale removal to provide a comprehensive molecular understanding of the early stages of these processes. Histological analysis of skin/scales revealed 3 days after scale removal re-epithelisation had occurred and the scale pocket had formed. In animals with scales removed, there was significant up-regulation of genes involved in cell cycle regulation, cell proliferation and adhesion, immune response and antioxidant activities. The expression profiles of the fasted animals centred on maintaining energy homeostasis. The utilisation of fasting as a treatment emphasised the competing whole animal physiological requirements with regard to barrier repair, infection control and energy homeostasis. Gene expression of sea bream (Sparus auratus) skin and scales was analysed in normal and treated animals. Three different treatments were applied: 1. scales removal at day 0 of the experiment; 2. unfed fish 7 days prior the start of the experiment; and 3. scales removal at day 0 of the experiment of unfed fish 7 days prior the start of the experiment. Fish were sampled at two different days: day 3 and day 7 after scale removal. Five individuals from control and experimental groups were analysed for both sampling days (3 and 7), resulting in a total of 40 samples analysed by microarray.

ORGANISM(S): Sparus aurata

SUBMITTER: Florbela Vieira

PROVIDER: E-GEOD-30717 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Skin healing and scale regeneration in fed and unfed sea bream, Sparus auratus.

Vieira Florbela A FA Gregório Silvia F SF Ferraresso Serena S Thorne Michael A S MA Costa Rita R Milan Massimo M Bargelloni Luca L Clark Melody S MS Canario Adelino V M AV Power Deborah M DM

BMC genomics 20111007

<h4>Background</h4>Fish scales are an important reservoir of calcium and phosphorus and together with the skin function as an integrated barrier against environmental changes and external aggressors. Histological studies have revealed that the skin and scales regenerate rapidly in fish when they are lost or damaged. In the present manuscript the histological and molecular changes underlying skin and scale regeneration in fed and fasted sea bream (Sparus auratus) were studied using a microarray 3 ...[more]

PMID: 21981800

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Project description:In Sparus aurata, seasonal temperature variations outside the normal thermal range, may trigger physiological responses leading to pathologies and death. In the present study two groups of wild sea bream were exposed for 21 days to two temperature regimes: 16 Â± 0.3 Â°C (control group) and 6.8 Â± 0.3 Â°C (cold-exposed group). Samples were collected during the acute phase (0, 6 and 24 hours after temperature drop) and upon chronic exposure (21 days). A comparative analysis of gene expression was conducted between S. aurata exposed to 6.8Â°C. Two groups of wild sea breams (120Â±16 g body weight; 21Â±1 cm total length) caught in Valle Bonello were kept in duplicate groups in four 5 m3 circular recycling tanks and acclimated over one month at 16 Â°C (Di Marco et al. 2010). Fish were then exposed for 21 days to two different temperature treatments: 16 Â± 0.3 Â°C (control groups) and 6.8 Â± 0.3 Â°C (cold groups). Both groups were fasted during the trial. Warm and cold water temperature conditions were maintained by supplying tanks with two separate re-circulating systems equipped with mechanical and biological filters. Cold water conditions were achieved by inducing two consecutive temperature drops: water was firstly gradually cooled from 16 Â°C to 12 Â°C at a rate of 1Â°C day -1 and then reduced from 12 Â°C to 6.8 Â°C within 24 h. This temperature was then kept for 21 days. Ten fish per treatment were sampled at different intervals during acute: 0 (T1), 6 (T2) and 24 hours (T3), and chronic exposure (21 days; T4) to low temperature. Fish were firstly anaesthetized in a solution of tricaine methanesulphonate (300 mg/l; MS 222, Finquel) in a 30-litre bucket and then euthanized by severing their spinal cord. Liver samples (about 30 mg) were collected from each specimen, incubated in RNA later at 4 Â°C for 24 h and then stored at -20 Â°C. Total RNA was extracted from tissue samples using the RNAeasy Mini Kit (Qiagen, Hilden, Germany). RNA concentration was determined using a UV-Vis spectrophotometer NanoDropÂ® ND-1000 (NanoDrop Technologies, Wilmington, USA). RNA integrity and quality was evaluated by means of Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA) and Expert software according to Ferraresso et al. (2008). RNA integrity number (RIN) index was calculated for each sample that provides a numerical assessment of RNA integrity. This value facilitates the standardization of the quality interpretation; only RNAs with RIN >8 were used for further microarray experiments. Once single RNA samples were extracted and measured, 3 pools (Pool_A consisting of four individuals, Pool_B and _C of three) per each sampling-time point were prepared. This was aimed at getting a sufficient statistical power which is reached by a minimum of three biological replicates per analysed point and at reducing microarray analysis cost. To perform DNA microarray analyses has been used the microarray platform GPL6467 (Ferraresso et al. 2008). Sample labelling and hybridization were performed according to the Agilent One-Color Microarray-Based Gene Expression Analysis protocol. For each of 24 liver pools 500 ng of total RNA were linearly amplified and labelled with Cy3-dCTP, using the Low RNA Input Linear Amplification Kit PLUS one-color (Agilent Technologies). A total of 24 Agilent 4 x 44K sea bream microarrays (12 samples from both control and cold groups) were then hybridized. An Agilent G2565BA DNA microarray scanner was used to scan arrays at 5 Âµm resolution, Feature Extraction Software 9.5.1 was then used to process and analyse array images. The software returns a series of spot quality measures in order to evaluate the goodness and the reliability of spot intensity estimates. Filtering and cyclic lowess normalization were performed using R statistical software (http://www.r-project.org/).

Dataset Information

Skin healing and scale regeneration in fed and unfed sea bream, Sparus auratus

Publications

Skin healing and scale regeneration in fed and unfed sea bream, Sparus auratus.

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