Análisis de la expresión de genes de respuesta inmune durante el desarrollo ontogénico de paralarvas de pulpo Octopus vulgaris criadas en cautividad

El pulpo común es una especie de gran importancia comercial, considerada como una especie emergente en acuicultura. En este trabajo se analizó el nivel de expresión de los genes inmunes TLR, C1q, Galectina, PGRP, LITAF, SERPIN, PRDX y Caspasa 3 mediante PCR cuantitativa (q-PCR) en embriones y paralarvas de O. vulgaris de edades 0, 10, 20 y 34 días. Adicionalmente, se infectaron paralarvas de 22 días con bacterias patógenas vivas Vibrio lentus y V. splendidus a 1h, 4h y 24h. El estudio del desarrollo del sistema inmune de estas paralarvas ayudará a identificar factores claves para la supervivencia y cultivo del pulpo común. Durante el desarrollo ontogénico, los embriones mostraron el menor nivel de expresión de PGRP, Caspasa 3 y PRDX. Por el contrario, C1q, Galectina y LITAF se observaron visiblemente expresados. C1q, TLR y SERPIN fueron los genes que presentaron mayor nivel de expresión en Pa0D. A partir de Pa10D se observó un notable incremento en la expresión de C1q, Galectina, PGRP y LITAF. La expresión de Caspasa3 se incrementó gradualmente desde Em. V. lentus y V. splendidus inducen un notable incremento de la expresión de C1q y PRDX entre 1h y 4h post infección. Sin embargo, durante las primeras horas de infección se observó una disminución de la expresión de Galectina, TLR, PGRP y LITAF. Particularmente la infección por V. lentus produjo una disminución de la expresión de SERPIN. Los resultados obtenidos sugieren que la capacidad del sistema inmune de reconocer patógenos y evitar infecciones es significativamente activo desde los estadíos de paralarvas recién eclosionadas. Sin embargo, se observó un aumento significativo de los genes seleccionados a partir de Pa10D.The common octopus is a species of high commercial interest and nowadays is considered as an emergent species in aquaculture. The mRNA expression level of immune-related genes (TLR, C1q, Galectin, PGRP, LITAF, SERPIN, PRDX and Caspase 3) was analyzed by Real time qPCR on embryos and paralarvae of O. vulgaris at age of 0, 10, 20 and 34 days. Additionally, paralarvae of 22 days were challenged with live Vibrio lentus and V. splendidus during a time course (1 h, 4 h and 24 h). This analysis will allow a better understanding of the developmental of immune system of these paralarvae, which will help to identify key factors for survival and culture of the common octopus. Regarding ontogeny, Em showed the lowest expression of PGRP, Caspase 3 and PRDX. In contrast, C1q, Galectin and LITAF were visibly expressed. C1q, TLR and SERPIN were highly expressed in Pa0D. A notably increase in the expression of C1q, Galectin, PGRP and LITAF began from Pa10D. Caspase 3 expression was gradually increased from Em. V. lentus and V. splendidus induced a notable expression of C1q and PRDX at 1 h and 4 h post infection (p. i.). Nevertheless, they markedly suppress the activation of Galectin, TLR, PGRP and LITAF during the first hours p.i.. Particularly, V. lentus suppressed the expression of SERPIN. The present results suggest that the ability of immune system to recognize pathogens and avoid infections is a priority in recently hatched paralarvae. However, a noticeable increase of gene expression was observed from Pa10D.0,000

Selection of reliable reference genes for RT-qPCR studies in Octopus vulgaris paralarvae during development and immune-stimulation

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COMPARATIVE STUDY OF AGE ESTIMATION IN WILD AND CULTURED Octopus vulgaris PARALARVAE. EFFECT OF TEMPERATURE

The common octopus Octopus vulgaris is a highly valuable species worldwide, but to understand its population dynamics and requirements under culture conditions, it is crucial to improve our knowledge about its planktonic stages. Previous studies validating daily beak growth increments in these stages allowed age estimation and comparison of wild and cultured paralarvae. We aimed to improve age estimations in captivity, addressing the effect of emperature and diet, to obtain an accurate estimation of age in wild specimens collected from the coast to the open ocean off NW Spain and Morocco. We analysed the beak growth increments of reared paralarvae at 14 and 21°C with 2 different crustacean prey taxa (Artemia and spider crab Maja brachydactyla zoeae) over 30 d. Daily increment deposition at 21°C was confirmed, whereas <1 increment d−1 was recorded at 14°C. The width of the reading area grew accordingly with age; therefore, this beak region may be suitable for age estimation. A general linear model (GLM) analysis showed that temperature and the interaction of age × temperature significantly influenced increment deposition, whereas diet did not. The number of growth rings recorded in wild paralarvae beaks ranged from 0−8 on the coast, 7−11 on the continental shelf and 2−28 in the open ocean. Corrected age estimates of wild paralarvae were obtained with the GLM using the mean temperaturas recorded in the wild, supporting the hypothesis that O. vulgaris leave the coastal area and develop in the open ocean transported by upwelling filaments.En prens