Molecular and Biochemical Methods Useful for the Epigenetic Characterization of Chromatin-Associated Proteins in Bivalve Molluscs

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dc.contributor.authorRivera-Casas, Ciro
dc.contributor.authorGonzalez-Romero, Rodrigo
dc.contributor.authorGarduño, Rafael A.
dc.contributor.authorCheema, Manjinder S.
dc.contributor.authorAusio, Juan
dc.contributor.authorEirin-Lopez, Jose M.
dc.date.accessioned2018-11-20T17:26:25Z
dc.date.available2018-11-20T17:26:25Z
dc.date.copyright2017en_US
dc.date.issued2017
dc.description.abstractBivalve molluscs constitute a ubiquitous taxonomic group playing key functions in virtually all ecosystems, and encompassing critical commercial relevance. Along with a sessile and filter-feeding lifestyle in most cases, these characteristics make bivalves model sentinel organisms routinely used for environmental monitoring studies in aquatic habitats. The study of epigenetic mechanisms linking environmental exposure and specific physiological responses (i.e., environmental epigenetics) stands out as a very innovative monitoring strategy, given the role of epigenetic modifications in acclimatization and adaptation. Furthermore, the heritable nature of many of those modifications constitutes a very promising avenue to explore the applicability of epigenetic conditioning and selection in management and restoration strategies. Chromatin provides a framework for the study of environmental epigenetic responses. Unfortunately, chromatin and epigenetic information are very limited in most non-traditional model organisms and even completely lacking in most environmentally and ecologically relevant organisms. The present work aims to provide a comprehensive and reproducible experimental workflow for the study of bivalve chromatin. First, a series of guidelines for the molecular isolation of genes encoding chromatin-associated proteins is provided, including information on primers suitable for conventional PCR, Rapid Amplification of cDNA Ends (RACE), genome walking and quantitative PCR (qPCR) experiments. This section is followed by the description of methods specifically developed for the analysis of histone and SNBP proteins in different bivalve tissues, including protein extraction, purification, separation and immunodetection. Lastly, information about available antibodies, their specificity and performance is also provided. The tools and protocols described here complement current epigenetic analyses (usually limited to DNA methylation) by incorporating the study of structural elements modulating chromatin dynamics.en_US
dc.description.reviewstatusRevieweden_US
dc.description.scholarlevelFacultyen_US
dc.description.sponsorshipThis material is based upon work supported by the National Science Foundation under Grant No. HRD-1547798. This NSF Grant was awarded to Florida International University as part of the Centers of Research Excellence in Science and Technology (CREST) Program. This is contribution number 40 from the Marine Education and Research Center in the Institute of Water and Environment at Florida International University. Additional support was provided by start-up funds from the College of Arts, Sciences and Education (CASE) at Florida International University (JE), the Natural Science and Engineering Research Council of Canada (NSERC) grant 43699-2012 (JA), and the Fundacion Ramon Areces (CR).en_US
dc.identifier.citationRivera-Casas, C.; Gonzalez-Romero, R.; Garduño, R.A.; Cheema, M.S.; Ausio, J.; & Eirin-Lopez, J.M. (2017). Molecular and biochemical methods useful for the epigenetic characterization of chromatin-associated proteins in bivalve molluscs. Frontiers in Physiology, 8, article 490. doi: 10.3389/fphys.2017.00490en_US
dc.identifier.urihttps://doi.org/10.3389/fphys.2017.00490
dc.identifier.urihttp://hdl.handle.net/1828/10336
dc.language.isoenen_US
dc.publisherFrontiers in Physiologyen_US
dc.subjectepigenetics
dc.subjectbivalves
dc.subjectchromatin
dc.subjecthistones
dc.subjectSNBPs
dc.subjectmethods
dc.subjectenvironment
dc.subject.departmentDepartment of Biochemistry and Microbiology
dc.titleMolecular and Biochemical Methods Useful for the Epigenetic Characterization of Chromatin-Associated Proteins in Bivalve Molluscsen_US
dc.typeArticleen_US

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