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dc.contributor.authorMantineo, Helene X.
dc.date.accessioned2021-05-15T15:53:47Z
dc.date.available2021-05-15T15:53:47Z
dc.date.issued2021-05-16
dc.identifier.citationMantineo, Helene X. (2021, May 16). Investigation of ciliated cell types and ciliary gene expression during heart regeneration in zebrafish (Danio rerio). Retrieved from:en_US
dc.identifier.urihttps://digitalrepository.wheatoncollege.edu/handle/11040/31685
dc.description119 leaves : illustrations.
dc.descriptionIncludes bibliographical references (leaves 105-109).
dc.description.abstractUnlike the adult mammalian heart, the adult zebrafish (Danio rerio) heart can regenerate after injury. In a series of regenerative processes, the zebrafish can regenerate their hearts after up to 20% of their ventricle apex is resected. The process of regeneration is directed via highly coordinated cell signaling pathways. Primary cilia act as directors of cell signaling pathways. Important ciliary signaling pathways have been implicated in zebrafish heart regeneration, but the direct link between regeneration and cilia has not been fully elucidated. Previous studies have found cilia present in the outer region of the regenerating zone during zebrafish heart regeneration. This led to the first hypothesis that cilia were present in the epicardium in zebrafish heart regeneration because the epicardium is believed to cover the outer region of the ventricle apex during regeneration. To test this hypothesis, the present study used immunofluorescence of transgenic zebrafish to label the different cell types of the regenerating heart. It was revealed that cilia were present in the outer region regenerating zone but not on the myocardium. There was indirect evidence that the cilia are not on the epicardium. Previous studies also found a change in abundance and length of cilia during zebrafish heart regeneration. This led to the second hypothesis that ciliary genes were differentially expressed during zebrafish heart regeneration. To test this hypothesis, the present study used DESeq2 in R to determine differential gene expression of ciliary genes. A total of 196 ciliary transcripts were found to be differentially expressed in at least one time point in zebrafish heart regeneration. Overall, the ciliated cell types and ciliary gene expression may provide further evidence of the importance of cilia in zebrafish heart regeneration. Continuing to gain insight the mechanisms of zebrafish heart regeneration may lead to clinical applications for human cardiac injury.en_US
dc.description.tableofcontentsIntroduction -- Cardiac Regeneration in Zebrafish -- Cilia -- Regeneration Pathways Connected with Cilia -- Objectives of the Present Study -- Methods -- Heart Surgeries, Collection and Sectioning -- Making Buffers for Immunofluorescence -- Phosphate Buffered Saline (PBS) -- Phosphate Buffered Saline with Triton X-100 as detergent (PBS-T) -- Blocking Buffer for Sea Urchin Embryos -- Blocking Buffer for Heart Sections -- Citrate Buffer -- Immunofluorescence -- Materials --Immunofluorescence of Sea Urchin Embryos -- Immunofluorescence of Heart Sections -- Imaging of samples in the ICUC -- Materials -- Widefield Microscopy -- Taking Pictures with SPOT -- Image Overlays in Photoshop -- Preparing for Confocal Imaging -- Confocal Imaging -- Materials -- Startup procedure for the Lillie Zeiss LSM 710 -- Setting Up the Microscope -- Scanning the Samples -- Shutdown procedure for the Lillie Zeiss LSM 710 -- Image Analysis Using Fiji -- Merging Colors -- Creating Z-projections -- Creating 3D Projections --Adjusting the Brightness/Contrast -- RNA Sequencing -- Animal Husbandry and tissue collection -- RNA preparation and sequencing -- Galaxy Processing -- Gene Expression Analysis -- Converting Transcript IDs -- DESeq2 Analysis -- Examining Ciliary genes -- Principal Components Analysis -- Heatmapping -- Results -- Sea Urchin Immunofluorescence -- EK Immunofluorescence -- tcf21 Immunofluorescence --cmlc2 Immunofluorescence -- Ciliary Gene Expression -- Discussion -- Cilia are present in the regenerating heart -- Antibodies were partially effective for labeling cilia -- Using transgenic zebrafish lines to investigate which cardiac cell types are ciliated -- Which cell types really are ciliated? -- Investigation of cilia gene expression during regeneration compared with ciliogenesis -- Significance of findings: implications for the role of cilia in heart regeneration
dc.language.isoen_USen_US
dc.publisherWheaton College. (Norton, Mass.)en_US
dc.subjectUndergraduate research.
dc.subjectUndergraduate thesis.
dc.subject.lcshMyocardium -- Regeneration.
dc.subject.lcshMyocardial infarction.
dc.subject.lcshZebra danio -- Anatomy -- Regeneration.
dc.subject.lcshCilia and ciliary motion -- Research.
dc.subject.lcshImmunofluorescence -- Research.
dc.titleInvestigation of ciliated cell types and ciliary gene expression during heart regeneration in zebrafish (Danio rerio)en_US
dc.typeThesisen_US


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