Biobanking of a Marine Invertebrate Model Organism: The Sea Urchin

Paredes, Estefanía

doi:10.3390/jmse4010007

Cited by 28 publications

(28 citation statements)

References 59 publications

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“…In this study, the high total sperm motility after thawing was obtained at 5%–20% DMSO, as well as fertilization rate. The results reported in this investigation was consistent with other studies that concentrations from 5% to 10% were found suitable for oyster, pearl oyster and sea urchin specimens (Hassan et al, ; Liu et al, ; Paredes, ).…”

Section: Discussionsupporting

confidence: 93%

Sperm cryopreservation of the noble Scallop Chlamys nobilis by a programmable freezing method: Effect of cryoprotectant

et al. 2019

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A study on Chlamys nobilis sperm cryopreservation by a programmable freezing method was conducted under laboratory condition. Four cryoprotectant agents (dimethyl sulfoxide [DMSO], methanol [MET], propanediol[PG] and ethylene glycol [EG]) and four concentrations (5%, 10%, 20% and 30%) were evaluated for their ability to retain sperm motility, movement characteristics and fertility. Results showed that cryopreserved sperm total motility produced by DMSO and MET at 5%, 10% and 20% were higher than other cryoprotectant treatment groups (CPA groups), as well as rapid sperm percentage. The curvilinear (VCL) and straight line (VSL) velocity produced by DMSO at 5% significantly higher than other CPA groups (p < 0.05), while no significant differences were found for average path (VAP) velocity. The lateral head displacement (ALH) in all CPA groups was similar and without significant difference (p > 0.05), as well as the beat‐cross frequency (BCF). A significant higher fertilization rate was produced in DMSO than that in MET at same concentration (p < 0.05), and no significant differences were found for differing concentrations of the same cryoprotectant (p > 0.05). Overall, 5%‐20% DMSO was more suitable for Chlamys nobilis sperm programmable cryopreservation when the calcium‐free Hanks’ balanced salt solution was used as the extender, and 10°C/min from 0°C to −80°C was used as freezing rate. The findings presented in this study will benefit conservation programs for Chlamys nobilis.

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Section: Discussionsupporting

confidence: 93%

Sperm cryopreservation of the noble Scallop Chlamys nobilis by a programmable freezing method: Effect of cryoprotectant

et al. 2019

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“…lividus is considered a keystone herbivore, able to transform communities dominated by macroalgae into barren areas thereby reducing biodiversity and altering ecosystem functions [ 35 , 36 ]. The sea urchin has been extensively used as an invertebrate model organism in developmental biology and in ecotoxicology studies to assess the effects of marine pollution on marine organisms [ 19 ].…”

Section: Discussionmentioning

confidence: 99%

“…Echinoids have been considered ideal models for monitoring marine environmental hazards [ 11 ], because they are often key herbivore species, having a major role in structuring and controlling macroalgal assemblages, thereby shaping the benthic seascape, and also playing an important role in coastal food webs throughout the world [ 12 – 16 ]. They have traditionally been used as model organisms to study reproduction and early cell differentiation, sperm-oocyte interactions and apoptosis [ 17 – 19 ]. These organisms have been also proposed as valuable bioindicators for detecting environmental perturbations [ 20 – 23 ].…”

Section: Introductionmentioning

confidence: 99%

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High-quality RNA extraction from the sea urchin Paracentrotus lividus embryos

et al. 2017

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The sea urchin Paracentrotus lividus (Lamarck, 1816) is a keystone herbivore in the Mediterranean Sea due to its ability to transform macroalgal-dominated communities into barren areas characterized by increased cover of bare substrates and encrusting coralline algae, reduced biodiversity and altered ecosystem functions. P. lividus is also an excellent animal model for toxicology, physiology and biology investigations having been used for more than a century as a model for embryological studies with synchronously developing embryos which are easy to manipulate and analyze for morphological aberrations. Despite its importance for the scientific community, the complete genome is still not fully annotated. To date, only a few molecular tools are available and a few Next Generation Sequencing (NGS) studies have been performed. Here we aimed at setting-up an RNA extraction method to obtain high quality and sufficient quantity of RNA for NGS from P. lividus embryos at the pluteus stage. We compared five different RNA extraction protocols from four different pools of plutei (500, 1000, 2500 and 5000 embryos): TRIzol®, and four widely-used Silica Membrane kits, GenElute™ Mammalian Total RNA Miniprep Kit, RNAqueous® Micro Kit, RNeasy® Micro Kit and Aurum™ Total RNA Mini Kit. The quantity of RNA isolated was evaluated using NanoDrop. The quality, considering the purity, was measured as A260/A280 and A260/230 ratios. The integrity was measured by RNA Integrity Number (RIN). Our results demonstrated that the most efficient procedures were GenElute, RNeasy and Aurum, producing a sufficient quantity of RNA for NGS. The Bioanalyzer profiles and RIN values revealed that the most efficient methods guaranteeing for RNA integrity were RNeasy and Aurum combined with an initial preservation in RNAlater. This research represents the first attempt to standardize a method for high-quality RNA extraction from sea urchin embryos at the pluteus stage, providing a new resource for this established model marine organism.

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“…There is a cryopreservation protocol described for P. lividus sperm (Fabbrocini et al, 2014) that yields good motility. Cryopreservation protocols already exist or are under development for different developmental stages for 10-14 different sea urchin species (embryos and sperm), and since sea urchins are a highly demanded model, more applications will probably be further developed soon using cryopreserved cells, including toxicology (Paredes, 2015a).…”

Section: Cryopreserved Echinoidsmentioning

confidence: 99%

The Use of Cryopreserved Biological Material for Water Quality Assessment

Paredes

Bellas

2019

Front. Mar. Sci.

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The stated aim of this perspective article is to present new developments and discuss future directions on the applications of cryopreserved organisms to marine water quality assessment. To facilitate this, the authors provide a background of essential knowledge of cryopreservation when applied to ecotoxicology, as well as, practical examples available in literature. An integrated approach with combined monitoring of chemical status plus measurements of biological effects has been recommended extensively by international institutions for the assessment of marine pollution. Among the available techniques, bioassays have been considered as sufficiently robust to be incorporated in marine pollution monitoring programs. However, the routine application of bioassays has also allowed the identification of one of the factors that limits a more extensive use of such biological methods: the availability of biological material throughout the year, regardless of natural spawning periods. A solution to this limitation is the application of cryopreservation techniques. Cryopreservation may, for instance, provide access to stable quality biological material when test species are out of the reproductive season, without the need for maintaining and conditioning organisms in the laboratory. It also guarantees access to a large variety of species that might not be available at the same time of the year and, on top of that, cryopreservation provides opportunities to laboratories that might not have the facilities to keep all these organisms in culture.

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Biobanking of a Marine Invertebrate Model Organism: The Sea Urchin

Cited by 28 publications

References 59 publications

Sperm cryopreservation of the noble Scallop Chlamys nobilis by a programmable freezing method: Effect of cryoprotectant

Sperm cryopreservation of the noble Scallop Chlamys nobilis by a programmable freezing method: Effect of cryoprotectant

High-quality RNA extraction from the sea urchin Paracentrotus lividus embryos

The Use of Cryopreserved Biological Material for Water Quality Assessment

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