Analysis of movement disorder-related genes following knockdowns of ANO10, WDR81, and VLDLR in zebrafish (DANIO RERIO)

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buir.advisorAdams, Michelle Marie
dc.contributor.authorŞengül, Göksemin Fatma
dc.date.accessioned2018-08-01T11:30:24Z
dc.date.available2018-08-01T11:30:24Z
dc.date.copyright2018-06
dc.date.issued2018-06
dc.date.submitted2018-07-17
dc.departmentGraduate Program in Neuroscienceen_US
dc.descriptionCataloged from PDF version of article.en_US
dc.descriptionThesis (M.S.): Bilkent University, Department of Neuroscience, İhsan Doğramacı Bilkent University, 2018.en_US
dc.descriptionIncludes bibliographical references (leaves 142-182).en_US
dc.description.abstractMovement Disorders are the neurological symptoms that cause alterations in normal motility, posture and muscle tone. Certain brain areas, such as the cerebellum, mediate correct motor control and functioning. When defects or congenital lesions occur in the cerebellum, neural disruption in motor coordination causes the development of a particular movement disorder known as cerebellar ataxia. The focus of this study was to examine how three genes of interest (ano10, wdr81, vldlr), contributing to multiple varieties of cerebellar ataxias, influence one another and other genes that are associated with this disorder. Mutations in vldlr and wdr81 are associated with Cerebellar Ataxia Mental Retardation Disequilibrium Syndrome type 1 and type 2 (CAMRQ1 and CAMRQ2), respectively, whereas mutations in the ano10 gene is responsible for the development of Autosomal Recessive Cerebellar Ataxia Type 3 (ARCA3). In this work, five key scientific findings were reported. Firstly, in silico analysis predicted a common Ca2+ activated Casein Kinase 2 (CK2) domain in the protein sequences of the genes of interest and also predicted a common interacting UBC protein. These predicted interactions, a common CK2 domain and a UBC interacting protein may explain the observed neurodegenerative phenotype in cerebellar ataxia. Secondly, the transcript level analysis (qPCR and RNASeq) of ano10a, wdr81 and vldlr using zebrafish embryos collected from early embryonic and late larval stages showed that the three genes were expressed relatively higher at 1 hpf, 2 hpf and 5 hpf developmental stages than others and may suggest their importance in developmental processes. Additionally, the comparison of the expression patterns of ano10a, wdr81 and vldlr during early embryogenesis indicated that three targeted genes were co-localized at diencephalon, midbrain (optic tectum) and cerebellum. These spatiotemporal results may restrict the involvement of these three genes selectively in early neurodevelopmental processes. Thirdly, this study also examined the expression level analysis of three targeted genes in 12 different adult tissues in a sexually dimorphic manner. Findings showed that genes of interest were expressed significantly higher at the eyes, brain and gonads (p-values < 0.05). Moreover, the gender specific examination in the 12 adult tissues revealed that ano10a and wdr81 expression differed significantly at eyes, gills, liver and gonads (p-values < 0.05) whereas, vldlr gene expression was significantly different at swim bladder and gonads in male and female individuals (p-values < 0.05). Fourthly, the clustergram analysis indicated that three genes of interest were grouped within close families with each other and 9 additional cerebellar ataxia associated genes and may imply that targeted genes alter functions in the converging pathways. Finally, results from the clustergram analysis helped to design and carry out a study knocking down the expression of ano10a, wdr81 and vldlr separately with MO antisense technology to examine the effect of the silenced mRNA on the expression levels of each other and 9 other highly correlated cerebellar ataxia-related genes. Single MO injections caused the significant upregulation of all investigated genes especially at 72 hours after/post injection (hpi) (p-values < 0.05) when ano10a transcript was silenced suggesting either an activated compensatory mechanism or activated alternative disease specific cascade molecules in response to its absence. Taken together, the outcomes of functional knockdowns can pave the way for the development of novel therapeutic targets using inhibitors or antagonists of activated cellular pathway components or the enhancers of downregulated genes to prevent or at least slow down the progression of not only cerebellar ataxia but also several other neurodegenerative disorders.en_US
dc.description.degreeM.S.en_US
dc.description.statementofresponsibilityby Göksemin Fatma Şengül.en_US
dc.embargo.release2021-06-28
dc.format.extentxviii, 205 leaves : illustrations (some color), charts ; 30 cm.en_US
dc.identifier.itemidB158689
dc.identifier.urihttp://hdl.handle.net/11693/47710
dc.language.isoEnglishen_US
dc.publisherBilkent Universityen_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.subjectZebrafish, Gene Expressionen_US
dc.subjectMovement Disordersen_US
dc.subjectCerebellar Ataxiaen_US
dc.subjectAno10en_US
dc.subjectWdr81en_US
dc.subjectVldlren_US
dc.subjectMorpholino Injectionen_US
dc.subjectGene Knockdownen_US
dc.subjectSexual Dimorphismen_US
dc.subjectIn Situ Hybridizationen_US
dc.subjectClustergram Analysisen_US
dc.subjectBioinformatics Analysis And qPCRen_US
dc.titleAnalysis of movement disorder-related genes following knockdowns of ANO10, WDR81, and VLDLR in zebrafish (DANIO RERIO)en_US
dc.title.alternativeZebrabalığında hareket bozukluğuyla ilişkilendirilen ANO10, WDR81 ve VLDLR genlerinin susturulduğu modelde incelenmesien_US
dc.typeThesisen_US

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