Martínez-Vázquez, J.M.

Pérez, M. (Montse)

English

Martínez-Vázquez, Juan Manuel

Pérez Rodríguez, Montse

Digital.CSIC

FOXL2 AS GENETIC SEX MARKER ON WRECKFISH Polyprion americanus

Martínez-Vázquez, J. M. (Juan Manuel)

Repositorio Institucional Digital del IEO

610FOXL2 AS GENETIC SEX MARKER ON WRECKFISH Ppolyprion americanus) BROODSTOCKS AND WILD CAUGHTS-00DUWtQH]9i]TXH]DQG03pUH]&HQWUR2FHDQRJUi¿FRGH9LJR6XELGDDO5DGLR)DUR9LJR6SDLQEmail: martinez.vazquez.jm@gmail.comIntroduction:recN¿VK Polyprion americanus) is a species with an increasing interest in European aquaculture, for its easy adaptation to captivity, their rapid growth rate in the ¿rst years of life, and a high price in the marNet. +owever, this species has a very late se[ual maturation and no apparent dimorphisms Eetween males and females, which dif¿cult the management of the EroodstocNs. In order to develop a genetic tool that allows identi¿cation of se[ in this species, this study analyzes the levels of e[pression of )2;/ gene, involved in ovarian differentiation in verteErates, in different individuals with varying degrees of gonadal maturation, for assess its usefulness of this gene as a sex marker.0aterial and 0ethods6amples of gonadal and gill tissues were analyzed, oEtained from wreck¿sh of different Eroodstocks estaElished in *alicia 6pain) and wild specimens landed at ¿sh markets. 7he set of samples covering the different stages of gonadal maturation, from immature juveniles to completely developed animals in their spawning period.7he samples were stored in 51$later, and then the 51$ were extracted with 7rizol and puri¿ed with chloroform and isopropanolethanol washings. )or expression analysis 57 q3&5 was performed with the kit %rilliant III 8ltra)ast 6<%5 *reen 43&5 0aster  0ix $gilent 7echnologies, using primers speci¿c for wreck¿sh )2;/ developed Ey 0ejuto et al.  7he experimental gene expression design was Eased on comparative analysis of the cycle value threshold &t), using %actin as the endogenous control.  7hese analyzes were performed in a 2ne6tep 5eal7ime 3&5 6ystem from $pplied Biosystems.Results and Discussion7he extraction and puri¿cation of R1$ from Eoth sample types from dead specimens at ¿sh markets and the living breeders) was achieved successfully, indicating the suitability of the method for processing of biological material. 3reliminary results that the gene )2;/ is already expressed in juveniles females in which no external signs of sexual maturation are appreciated &t mean  ,,), and where the dissection of the gonads in the case of dead specimens) is necessary for sex determination. 7his early expression of )2;/ has already been observed in other teleost such as 1ile tilapia :ang et al., ). Its presence in wreck¿sh could be useful as a genetic tool to differentiate males from females in aquaculture stocks, especially given that this species reaches sexual maturity around  years of life 3eleteiro and Bruzyn 2014), and whose only external difference between the sexes is a small variation in the structure of gonopore appreciable only during the high accretion gonad phases in each breeding season (Peleteiro pers. com.).It is necessary to continue these expression analysis in order to consolidate this gene as a sex marker, and to assess whether expression levels in females can be used to infer what is their state of gonadal development. If so, would entail a less traumatic substitute for  the animal of the gonadal biopsy techniques by cannulation used for this purpose, resulting in lower handling of animals and a greater welfare in their culture conditions.ReferencesMejuto, A., Presa P., Pérez, M., 2012. B~squeda de genes implicados en la determinaciyn sexual en especies no consolidadas en acuicultura. 9igo. Departamento de Bioqutmica, *enptica e Inmunologta. 8niversidad de 9igoPeleteiro J.B., Bruzón, M.A. 2014. &ultivo de serrinidos: el mero y la cherna. In: 'iYersi¿caciyn Ge las especies en la piscicultura marina española. I.E.2., )undaciyn 2.E.6.$ (Eds.), pp: 201 241.Wang, S., Kobayashi, T., Zhou, L-V., Prasanth,B., Ijiri S., Sakai F., Okubo K., Morohashi K., Nagahama Y. 2004. )oxl2 8pRegulates $romatase *ene 7ranscription in a )emale 6peci¿c 0anner by Binding to the Promoter as well as interacting with Ad4 Binding Protein/Steroidgenic Factor 1. Mol. Endocrin. (21) 3: 712-725

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FOXL2 AS GENETIC SEX MARKER ON WRECKFISH Polyprion americanus

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