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

García-Fernández, Pablo; Castellanos-Martínez, Sheila; Iglesias, J.; Otero, Juan José; Gestal, Camino

doi:10.1016/j.jip.2016.06.003

Cited by 15 publications

(14 citation statements)

References 28 publications

(20 reference statements)

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“…spring All tissues and Mantle and Foot, autumn Body and Mantle, spring + autumn All tissues, Body and Mantle and Foot). This agrees with results reported in larval and adult cephalopods [ 20 , 23 ]. In Mytilus edulis and Haliotis discus EF1α could be identified as a stable reference gene [ 21 , 22 ].…”

Section: Discussionsupporting

confidence: 93%

“…It is also one of the genes that displayed stable expression across seasons. However it did not behave ideally in all data sets, a result that agrees with differences in BACT expression between tissues and treatments in other molluscs [ 20 , 21 , 22 , 23 , 24 ]. Our results also identified the traditional housekeeping gene UBI as a good candidate reference gene in multiple data sets in both spring, autumn and spring + autumn (e.g.…”

Section: Discussionsupporting

confidence: 59%

“…To date, only a handful of molluscan species have had reference genes tested for stable expression [ 20 , 21 , 22 , 23 , 24 ], and in C . nemoralis this has never been performed.…”

Section: Introductionmentioning

confidence: 99%

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Identification and validation of reference genes for qPCR in the terrestrial gastropod Cepaea nemoralis

Affenzeller

Cerveau

2018

PLoS ONE

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Identifying and understanding mechanisms that generate phenotypic diversity is a fundamental goal of evolutionary biology. With a diversity of pigmented shell morphotypes governed by Mendelian patterns of inheritance, the common grove snail Cepaea nemoralis (Linnaeus, 1758) has been a model for evolutionary biologists and population geneticists for decades. However, the genetic mechanisms by which C. nemoralis generates this pigmented shell diversity remain unknown. An important first step in investigating this pigmentation pattern is to establish a set of validated reference genes for differential gene expression assays. Here we have evaluated eleven candidate genes for reverse transcription quantitative polymerase chain reaction (qPCR) in C. nemoralis. Five of these were housekeeping genes traditionally employed as qPCR reference genes in other species, while six alternative genes were selected de novo from C. nemoralis transcriptome data based on the stability of their expression levels. We tested all eleven candidates for expression stability in four sub-adult tissues of C. nemoralis: pigmented mantle, unpigmented mantle, head and foot. We find that two commonly employed housekeeping genes (alpha tubulin, glyceraldehyde 3-phosphate dehydrogenase) are unsuitable for use as qPCR reference genes in C. nemoralis. The traditional housekeeping gene UBIquitin on the other hand performed very well. Additionally, an RNA-directed DNA polymerase (RNAP), a Potassium Channel Protein (KCHP) and a Prenylated Rab acceptor protein 1 (PRAP), identified de novo from transcriptomic data, were the most stably expressed genes in different tissue combinations. We also tested expression stability over two seasons and found that, although other genes are more stable within a single season, beta actin (BACT) and elongation factor 1 alpha (EF1α) were the most reliable reference genes across seasons.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionsupporting

confidence: 59%

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Identification and validation of reference genes for qPCR in the terrestrial gastropod Cepaea nemoralis

Affenzeller

Cerveau

2018

PLoS ONE

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“…Incidence of diseases, several of them caused by bacteria, is one of the most important problems which avoid obtaining a successfully production in the aquaculture of cephalopods or its suitable maintenance in captive conditions (García--Fernández et al 2016;Sykes and Gestal, 2014).…”

Section: Introductionmentioning

confidence: 99%

Bacteria-Affecting Cephalopods

Farto

Fichi

Gestal

et al. 2019

Handbook of Pathogens and Diseases in Cephalopods

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Bacterial pathogens contribute to obtain an unsuccessful production of cephalopods. An updated overview of the knowledge of these pathogens must be a valuable tool to improve their aquarium maintenance and aquaculture. The present work provides a description of the main bacterial pathogens associated with larval stages of cultured Octopus vulgaris, and juvenile and adults of several cephalopods. Vibrio species, reported with ability to cause vibriosis in aquaculture, are the main bacteria associated with skin lesions in adults. Different species of Pseudomonas and Aeromonas, among others, have also been detected. Furthermore, gram-positive bacteria such as Bacillus have been also described. Among them, V. alginolyticus, V. carchariae, V. parahaemolyticus, V. splendidus and V. lentus have also been isolated from sterile organs or fluids of animals and their potential as invaders proved. However, only V. alginolyticus or V. lentus has the ability to cause lesions, and, in addition, the last one is proved as the causative agent of death in octopuses. Other organs such as eyes of squids are also colonized by Vibrio species or Micrococcus sp., and recently Photobacterium swingsii and Lactococcus garvieae have been reported associated with a retrobulbar lesion in octopus. Rickettsial-like organisms (RLO) are also detected in the gills of the octopus, having a detrimental effect on the respiratory gaseous exchange of the animals. Cultures of octopus paralarvae show a genetically diverse community comparable to those reported previously from other marine hatcheries. Bacteria included in the Splendidus clade is the dominant group in all conditions, except in one of them, where V. alginolyticus, V.

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“…; García‐Fernández et al . , ). Most emerging disease problems in mollusc are associated with species for which there is negligible reference material of normal and abnormal conditions.…”

Section: Larviculture Challengesmentioning

confidence: 99%

Early life stage bottlenecks of carnivorous molluscs under captivity: a challenge for their farming and contribution to seafood production

Uriarte

Astorga

Navarro

et al. 2018

Reviews in Aquaculture

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This work brings together the view of different specialists in the areas of larviculture, physiology, ecology, nutrition and animal health, regarding how to deal with the aquaculture farming of species with complex life cycles in a multidisciplinary way, using as models the octopus and the muricid C. concholepas, with the aim of reducing the gap between the experimental and the industrial culture of species that are relevant for the diversification of aquaculture, particularly in Chile. Although these species are similar in their difficulty to reach the terminal planktonic phase prior to juvenile, they differ in the bottlenecks they have to overcome to reach it. Relevant aspects of study to achieve juvenile production from early life stages rearing, whether for repopulation or for ongrowing, are as follows: (i) Replacement or supplementation of live diets with inert diets to achieve significant survival values over the first stages of life. (ii) Physiological approaches to establish cultivation conditions evaluating the individual responses to several rearing conditions, specially the interaction between temperature, dissolved oxygen and acidity. Studies of urgent character due to the global warming scenario. (iii) Genomic studies associated with the effect of ontogenetic development, environment, health and nutrition on gene expression to understand, in an integrated way, the key processes for the development and growth of immature stages. (iv) Studies on the control of reproduction, the quality control of the ova, the genetic structure of reproductive populations and the characterization of diseases are also necessary to achieve efficient hatchery technologies.

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

Cited by 15 publications

References 28 publications

Identification and validation of reference genes for qPCR in the terrestrial gastropod Cepaea nemoralis

Identification and validation of reference genes for qPCR in the terrestrial gastropod Cepaea nemoralis

Bacteria-Affecting Cephalopods

Early life stage bottlenecks of carnivorous molluscs under captivity: a challenge for their farming and contribution to seafood production

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