(D) (i) Top panel, control embryos at different developmental stages. (ii) Relative change in levels of Myo1C ±PClP, 1-4 and 64 cell stages, control grey, PClP black. Control (top panel) and with PClP (bottom panel), GAPDH used as loading control. (C) (i) Western blot for Myo1C relativelevels at 1-4 and 64 cell stages. For Myo1Cb, control (top panel) and with PClP (bottom panel), were compared at 1-4 and 64 cells for cDNA and RNA templates. For Myo1Ea&b, control (top panel) and with PClP (bottom panel), were compared at 1-4 and 64 cells stages for cDNA and RNA templates, (B) Semiquantitative RT-PCR profiles from cDNA and RNA templates. (A) Semiquantitative RT-PCR profiles from cDNA and RNA templates. Schematic representation of Myo1 domain structures. Inhibition of Myo1 arrests cell division and affected blastomere shapeof Zebrafish embryos. Therefore, our results demonstrate a potential role for Myo1 in the maintenance and formation of furrow, blastodisc morphology, cell-division and LD organization within the blastodisc during early embryogenesis. All these results above are effects of Myo1 inhibition exclusively Myo2 inhibition by blebbistatin does not show such phenotypes. In addition to cell division and furrow formation, inhibition of Myo1 has a drastic effect on the dynamics and distribution of lipid droplets (LDs) in the blastodisc near the cleavage furrow. Myo1 appears to be crucial for maintaining the blastodisc morphology and the actin cortex thickness. We found that the blastodisc of the embryo, which contains a thick actin cortex (~13 μm), is loaded with cor-tical Myo1. While Myo1 isoforms appears to be important for both the formation and the maintenance of cleavage furrows, Myo2 is required only for the formation of furrows. Inhibition of Myo1 (by PClP) and Myo2 (by Blebbistatin) lead to arrest in cell division. Our results indicate a unique involvement of Myo1 in early development of Zebra-fish embryos. We have studied the effect of Myo1 inhibitor PClP in 1–8 cell Zebrafish embryos. Myosin-1 (Myo1) represents a mechanical link between the membrane and actin-cytoskele-ton in animal cells.
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