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Browsing by Subject "Fetal Alcohol Spectrum Disorder (FASD)"
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Item Effect of Curcuminoids in Turmeric on Developing Zebrafish Treated with Ethanol(Office of the Vice Chancellor for Research, 2016-04-08) Connors, Craig; Mohammed, Arooj; Muralidharan, Pooja; Sarmah, Swapnalee; Marrs, James; Marrs, Kathleen A.; Chism, GradyThis experiment was designed with the intention of determining whether turmeric could act as a rescue agent to prevent or mitigate the extent of Fetal Alcohol Spectrum Disorder (FASD) caused by early ethanol exposure using zebrafish as a model system. A range of turmeric concentrations were made from a stock solution of turmeric dissolved in ethanol (1mg turmeric in 5mL ethanol). The active agents in turmeric are the curcuminoids: Curcumin, Desmethoxycurcumin, and Bisdemethoxycurcumin. The curcuminoids concentration was estimated using liquid chromatography. These agents were present in the turmeric stock solution at the following concentrations: Bisdemethoxycurcumin: 36.6 +/- 0.1 ug/mL, Desmethoxycurcumin: 43.4 +/- 0.1 ug/mL, and Curcumin: 124.1 +/- 0.2 ug/mL. Untreated zebrafish embryos were placed in embryo medium, ethanol treated embryos in 100mM ethanol containing embryo medium, and turmeric co-supplemented medium with differing concentrations of turmeric. Since the turmeric stock solution was dissolved in ethanol, the concentration of ethanol was kept at a constant 100mM ethanol and the amount of turmeric solution added. The concentrations of the test plates were then based on this solution and made to be 100 mM ethanol and 1.16 uM curcuminoids, 100 mM ethanol and 1.74 uM curcuminoids, and 100 mM ethanol and 2.32 uM curcuminoids. The developing embryos were treated with the turmeric solution and/or ethanol during 2-24 hours post fertilization (hpf). These embryos were imaged at 72 hpf and their body length and eye diameter were measured. The embryos supplemented with curcuminoids showed a significant rescue effect on the body length and eye diameter compared to ethanol treated embryos. This indicates that the curcuminoids acted as a rescue agent to reduce the effects that are typical of FASD in developing zebrafish.Item Ethanol-Induced Defects on Zebrafish Retinal Development: Rescue by Nutritional Supplements(Office of the Vice Chancellor for Research, 2015-04-17) Muralidharan, P.; Sarmah, S.; Marrs, J. A.Fetal Alcohol Spectrum Disorder (FASD), a result of prenatal alcohol exposure, produces a wide range of developmental defects including severe ocular defects that include microphthalmia, optic nerve hypoplasia, scotopic vision loss and coloboma. The zebrafish FASD model recapitulates many defects seen in human patients. Ethanol exposure (100 and 150 mM) during early development (midblatula transition through somitogenesis, 2-24 hours post fertilization, hpf) produced severe ocular defects including microphthalmia, optic nerve hypoplasia and photoreceptor differentiation defect. Examining specific terminal differentiation markers showed ethanol-induced defects in differentiation of most retinal cell types. Ethanol exposure altered gene expression of critical transcription factors. Increased cell death accounted for the small eye phenotype, and the retina responds with increased proliferation in the outer nuclear layer, inner nuclear layer, and ciliary marginal zone (CMZ). Ethanol treated retinas showed an expanded CMZ and cell cycle exit defects of the photoreceptor cells. In order to examine progenitor cell populations and differentiation defects in the ethanol treated retinal cells, specific markers for retinal stem, precursor and progenitor cell populations were examined. While control retinas showed terminally differentiated photoreceptors at 72 hpf, ethanol treated retinas expressed immature and nascent photoreceptor markers in cell populations undergoing proliferation. Nutrient co-supplement with retinoic acid (RA) or folic acid (FA) with ethanol during 2-24 hpf rescued photoreceptor differentiation and optic nerve defects. Competitive inhibition of RA synthesis by ethanol was hypothesized by Duester (1991), and rescue of ethanol-induced retinal defects suggest an effect on RA levels in the developing retina. Treatment with RA inhibitors produced retinal defects similar to ethanol-treated embryos. Interestingly, RA supplementation (24-48 hpf and 48-72 hpf) following ethanol treatment (2-24 hpf) restored photoreceptor differentiation suggesting RA provides a critical signal for precursor cell differentiation. In contrast, post-treatment with FA, did not restore retinal cell differentiation. FA functions as a critical component of one-carbon metabolism and can influence histone- and DNA-methyl transferase activities. Molecular mechanisms underlying disruption of cell cycle exit and FA rescue of ethanol-induced defects are being actively studied.