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Castelló de la Plana, Spain

Milan M.,University of Padua | Milan M.,University of Florence | Coppe A.,University of Padua | Reinhardt R.,Max Planck Institute for Molecular Genetics | And 8 more authors.
BMC Genomics | Year: 2011

Background: The Manila clam, Ruditapes philippinarum, is one of the major aquaculture species in the world and a potential sentinel organism for monitoring the status of marine ecosystems. However, genomic resources for R. philippinarum are still extremely limited. Global analysis of gene expression profiles is increasingly used to evaluate the biological effects of various environmental stressors on aquatic animals under either artificial conditions or in the wild. Here, we report on the development of a transcriptomic platform for global gene expression profiling in the Manila clam.Results: A normalized cDNA library representing a mixture of adult tissues was sequenced using a ultra high-throughput sequencing technology (Roche 454). A database consisting of 32,606 unique transcripts was constructed, 9,747 (30%) of which could be annotated by similarity. An oligo-DNA microarray platform was designed and applied to profile gene expression of digestive gland and gills. Functional annotation of differentially expressed genes between different tissues was performed by enrichment analysis. Expression of Natural Antisense Transcripts (NAT) analysis was also performed and bi-directional transcription appears a common phenomenon in the R. philippinarum transcriptome. A preliminary study on clam samples collected in a highly polluted area of the Venice Lagoon demonstrated the applicability of genomic tools to environmental monitoring.Conclusions: The transcriptomic platform developed for the Manila clam confirmed the high level of reproducibility of current microarray technology. Next-generation sequencing provided a good representation of the clam transcriptome. Despite the known limitations in transcript annotation and sequence coverage for non model species, sufficient information was obtained to identify a large set of genes potentially involved in cellular response to environmental stress. © 2011 Milan et al; licensee BioMed Central Ltd.

Garcia S.,Centro IFAPA Agua del pino | Domingues P.,Centro IFAPA Agua del pino | Navarro J.C.,Institute Acuicultura Of Torre La Sal Iats | Hachero I.,Centro IFAPA Agua del pino | And 2 more authors.
Aquaculture Nutrition | Year: 2011

Effects of two binders (gelatine and alginate) were tested on growth, survival, partial energy balance and lipid composition of mantle and digestive gland (DG) of Octopus vulgaris. The three diets tested were given as follows: CON, (Loligo gahi) as control, GEL, composed of squid paste (L. gahi) (300gkg-1), fish hydrolyse CPSP® (100gkg-1) and fish meal (500gkg-1), agglutinated with 100gkg-1 of gelatine and ALG, composed of squid paste (L. gahi) (300gkg-1), fish hydrolyse CPSP® (100gkg-1) and fish meal (500gkg-1), all agglutinated with 100gkg-1 of alginate. Growth rates were 13.7±2.1, 2.1±2.8 and -2.4±2.9gkg-1bwday-1, for octopuses fed CON, GEL and ALG diets, respectively. DGs of octopuses had higher concentrations of fatty acids (FA) than the mantle. DG of animals fed CON had higher concentrations of FAs than those fed the artificial diets. Energetic balance demonstrated that physiologically useful energy for maintenance E(B) was affected by type of diet, with negative values of E(B) in animals fed ALG and positive (85 and 154kJkg-1day-1) in octopuses fed GEL and CON, respectively. The ALG diet did not cope with the physiological requirements for octopus growth. © 2010 Blackwell Publishing Ltd.

Leite R.B.,University of Algarve | Leite R.B.,Instituto Gulbenkian Of Ciencia | Milan M.,University of Padua | Coppe A.,University of Padua | And 8 more authors.
BMC Genomics | Year: 2013

Background: The Grooved Carpet shell clam Ruditapes decussatus is the autochthonous European clam and the most appreciated from a gastronomic and economic point of view. The production is in decline due to several factors such as Perkinsiosis and habitat invasion and competition by the introduced exotic species, the manila clam Ruditapes philippinarum. After we sequenced R. decussatus transcriptome we have designed an oligo microarray capable of contributing to provide some clues on molecular response of the clam to Perkinsiosis.Results: A database consisting of 41,119 unique transcripts was constructed, of which 12,479 (30.3%) were annotated by similarity. An oligo-DNA microarray platform was then designed and applied to profile gene expression in R. decussatus heavily infected by Perkinsus olseni. Functional annotation of differentially expressed genes between those two conditionswas performed by gene set enrichment analysis. As expected, microarrays unveil genes related with stress/infectious agents such as hydrolases, proteases and others. The extensive role of innate immune system was also analyzed and effect of parasitosis upon expression of important molecules such as lectins reviewed.Conclusions: This study represents a first attempt to characterize Ruditapes decussatus transcriptome, an important marine resource for the European aquaculture. The trancriptome sequencing and consequent annotation will increase the available tools and resources for this specie, introducing the possibility of high throughput experiments such as microarrays analysis. In this specific case microarray approach was used to unveil some important aspects of host-parasite interaction between the Carpet shell clam and Perkinsus, two non-model species, highlighting some genes associated with this interaction. Ample information was obtained to identify biological processes significantly enriched among differentially expressed genes in Perkinsus infected versus non-infected gills. An overview on the genes related with the immune system on R. decussatus transcriptome is also reported. © 2013 Leite et al.; licensee BioMed Central Ltd.

Espigares F.,Institute Acuicultura Of Torre La Sal Iats | Zanuy S.,Institute Acuicultura Of Torre La Sal Iats | Gomez A.,Institute Acuicultura Of Torre La Sal Iats
Biology of Reproduction | Year: 2015

Kisspeptins are key players in the neuroendocrine control of puberty and other reproductive processes in mammals. Several studies have demonstrated that the KISS/GPR54 system is expressed by gonadotrophs, but in vitro studies assessing the direct stimulatory effects of kisspeptin on gonadotropin secretion in the pituitary have provided conflicting results. In this study, we investigated whether kisspeptin directly influences the reproductive function of sea bass pituitary. First, the highly active peptides Kiss1-15 and Kiss2-12 were used to stimulate dispersed sea bass pituitary cells obtained from mature males. Our results show that, first, Kiss2-12 induced luteinizing hormone (Lh) and follicle-stimulating hormone (Fsh) release, whereas Kiss1-15 had no effect on gonadotropin secretion at full spermiation stage. Second, the distribution and nature of Kiss2 and its potential interactions with the gonadotropin-releasing hormone 1 (Gnrh1) system in the pituitary were analyzed using dual fluorescence immunohistochemistry. Kiss2 cells were found in the proximal pars distalis and colocalized with gonadotropin-immunoreactive cells. In summary, our results provide, for the first time in a teleost species, functional and neuroanatomical evidence that Kiss2 may act through different routes to directly modulate the activity of gonadotrophs, either as a hypophysiotropic neuropeptide or as an autocrine/paracrine factor. © 2015 by the Society for the Study of Reproduction, Inc.

Espigares F.,Institute Acuicultura Of Torre La Sal Iats | Carrillo M.,Institute Acuicultura Of Torre La Sal Iats | Gomez A.,Institute Acuicultura Of Torre La Sal Iats | Zanuy S.,Institute Acuicultura Of Torre La Sal Iats
Biology of Reproduction | Year: 2015

Some teleost species, including European sea bass, harbor two different kisspeptin coding genes: kiss1 and kiss2. Both genes are expressed in the brain, but their differential roles in the central control of fish reproduction are only beginning to be elucidated. In this study, we have examined the effects of intracerebroventricular injections of the highly active sea bass peptides Kiss1-15 and Kiss2-12 on spermiating male sea bass. Physiological saline, Kiss1-15, or Kiss2-12 was injected into the third ventricle. To establish the gene expression cascade involved in the action of kisspeptins, the expression of the two sea bass kisspeptin receptor genes (kiss1r and kiss2r) and the three sea bass Gnrh genes (gnrh1, gnrh2, and gnrh3) were analyzed in the forebrain-midbrain and the hypothalamus. In addition, the protein levels of hypothalamic and pituitary Gnrh1 were measured. Blood samples were collected at different times after injection to analyze the effects of kisspeptins on the release of gonadotropins (Lh and Fsh) and androgens (testosterone and 11-ketotestosterone). The present results provide the first evidence that the effects of Kiss2 on central regulation of reproductive function involve the neuroendocrine areas of the forebrain-midbrain in teleost fish. The marked effect of Kiss2 on kiss2r and gnrh1 expression in the forebrain-midbrain and on Gnrh1 release suggest that this neuronal system is involved in the neuroendocrine regulation of gonadotroph activity. This hypothesis was confirmed by a surge of plasma Lh in response to Kiss2, which presumably has a strong stimulatory effect on testosterone release, and thus on sperm quality parameters.

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