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(E,F) The same results presented as a percentage of GFP+ cells/parent population also show no significant differences upon pyriproxyfen exposure among both gfap+ cells and nestin+ cells (p > 0.05). (C,D) The number of gfap+ cells and nestin+ counted per 100 beads was not significantly different upon pyriproxyfen exposure when compared to vehicle-treated embryos (p > 0.05). The beads are completely separated from GFP+ cells on a FSC/SSC dot plot (lower panel, arrow).
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The fluorescent beads were distinguishable using a 561 nm laser with a 610/20 BP filter and a 405 nm laser with a 525/50 BP filter. (B) The GFP+ cell population ( embryos) was distinguishable using a 488 nm laser with a 530/30 BP filter (upper panel). 100 beads were counted by flow cytometry and the number of GFP+ cells was counted simultaneously. (A) Whole 24 hpf transgenic embryos ( or were dissociated into single-cells in a medium containing a 1,000 fluorescent beads. Neural stem cell populations are not affected by pyriproxyfen exposure. sc.: spinal cord hb.: hindbrain m/h: midbrain/hindbrain boundary mb.: midbrain fb.: forebrain ot.: optic tectum tv.: tectum ventricle ob.: olfactory bulb. (D,E) Quantification of embryonic (2 dpf) (D) and larval (7 dpf) (E) brain volumes shows no significant differences between vehicle-treated and pyriproxyfen-treated embryos (p > 0.05).
#Vani rani today episode 1093 software#
(C) 3D-volume reconstruction of embryos using Imaris software (Bitplane) and confocal microscopy. The morphology of the larval brain is not affected by any of the doses. Of note is that none of the embryos treated with a dose of 1 μ g/ml survived past 6 dpf. (B) Transgenic 7 dpf larvae treated with vehicle, 0,005 μ g/ml, 0,01 μ g/ml, 0,1 μ g/ml pyriproxyfen were imaged under a confocal microscope. The morphology of the embryonic brain is not affected by any of the doses. (A) Transgenic 2 dpf embryo treated with vehicle, 0,005 μ g/ml, 0,01 μ g/ml, 0,1 μ g/ml and 1 μ g/ml pyriproxyfen were imaged under a confocal microscope. Our data indicate that pyriproxyfen does not affect central nervous system development in zebrafish, suggesting that this larvicide on its own, may not be correlated with the increase in microcephaly cases reported recently.Įmbryonic and larval brain volumes are not affected by pyriproxyfen treatment. Interestingly, though lethal at very high doses, pyriproxyfen did not cause brain malformation nor any significant changes in the number of observed stem cells in the developing central nervous system. Using zebrafish transgenic lines expressing GFP in different cell populations (elavl3 in newborn neurons, gfap and nestin in neural stem cells), we focused on the analysis of whole embryonic brain volume after confocal 3D-reconstruction and the quantification of purified neural stem cells during early neurodevelopment by FACS-cell sorting from whole in vivo embryos. As a result, we aimed at addressing the effects, if any, of pyriproxyfen on neurodevelopment in the zebrafish embryo as a vertebrate model. The effects of this compound on neurodevelopment have not yet been addressed specifically in vertebrates.
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Although the zika virus (ZIKV) has now been strongly correlated with emerging cases of microcephaly in the Americas, suspicions have been raised regarding the use of pyriproxyfen, a larvicide that prevents mosquito development, in drinking water.
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