Browsing by Subject "Morpholino"

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Open Access
The characterization and potential functional role of wdr81, a novel zebrafish gene, associated with cerebellar ataxia, mental retardation and dysequilibrium syndrome (camrq) in humans
(2016-04) Doldur Ballı, Füsun
Cerebellar ataxia, mental retardation and dysequilibrium syndrome (CAMRQ) is a neurodevelopmental disorder. The gene encoding WD repeat containing protein 81 (WDR81) was reported to be associated with CAMRQ2 [MIM 610185]. Human and mouse studies indicated the potential importance of WDR81 in neurodevelopment. The first aim in this study was to characterize the transcript and to reveal the expression profile of wdr81 in zebrafish. The second aim was to perform the initial characterization of wdr81 morphants. In silico analysis indicated that the conserved domains are shared in human, mouse and zebrafish orthologous proteins, implying a conserved function of WDR81 in three species. The characterization of the transcript revealed that wdr81 possessed one ORF and one 5’UTR structure. The predicted sequence for 3’UTR was confirmed along with detection of some variants and an insertion site in samples from ten developmental timepoints and in several adult tissues. This region was not detected in kidney, intestine and gills, which might be pointing out an alternative polyadenylation event. wdr81 appeared to be maternally supplied. 5 hpf and 18 hpf were detected as crucial timepoints regarding wdr81 expression. Expression of wdr81 was found to be increased in the eye and brain regions at 18 hpf and 48 hpf. wdr81 was found to be ubiquitously expressed in the adult zebrafish. The expression of wdr81 in the adult brain and eye was detected in several regions including retinal layers, presumptive Purkinje cells and some proliferative zones. The splice blocking morpholino which targets the exon 2-intron 2 junction of wdr81 worked at 3 tested doses; 2 ng, 4 ng and 8 ng. The effect of the wdr81 morpholino was detected to add the intron, which is downstream of the target exon, to the transcript and introduce a stop codon. Preliminary results indicated a significant reduction in the head sizes at a ratio of 3.88% (p:0.027) in the wdr81 morphant group compared to uninjected group and gbx2 expression was observed to be higher in wdr81 morphants compared to the control groups. In short, findings of this study emphasize the significance of wdr81 in neurodevelopment and suggest a potential role in neuronal proliferation. This study also serves as a basis for future functional studies.
Open Access
Zebrafish aging models and possible interventions
(InTechOpen Press, 2018) Birand, Dilan Ç.; Erbaba, Begün; Özdemir, Ahmet T.; Kafalıgönül, Hulusi; Adams, Michelle; Bozkurt, Y.
Across the world, the aging population is expanding due to an increasing average life expectancy. The percentage of elderly over the age of 65 is expected to be more than 15% of the total world population by 2025. As the lifespan increases, there will be a need for maintaining a healthy state for these individuals. Our current knowledge on types and durations of potential anti-aging therapies is quite limited. Recently the zebrafish has emerged as a promising model for understanding the cognitive and neurobiological changes during aging, as well as its use with potential anti-aging interventions. Like humans this model organism ages gradually, displays similar behavioral properties and social characteristics, and in addition, there is a wealth of molecular and genetic tools to uncover the cellular mechanism that contribute to age-related cognitive declines. Drug effect and toxicity can be easily tested in the zebrafish. Therefore, this animal model can provide information about potential therapies that could be translated directly into human populations or provide a more focused treatment direction for testing in other mammalian animal models. The zebrafish will be a powerful tool for uncovering the mysteries of the aging brain.