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Proteins which are Linked with Swiss Webster Mouse Limb Teratogenesis as the Effects of Methoxyacetic Acid (MAA) Treatment

Aceng Ruyani, Sri Sudarwati, Lien A. Sutasurya, Sony H. Sumarsono

 


Abstract. The analysis of proteins, which are linked with limb teratogenesis as the effects of MAA treated in Swiss Webster mouse has been investigated. A single dose of MAA 10 mmol/kg body weight was given by gavage on gestation day 11, whereas the control group were administered sterilized distilled water. Pregnant mice were sacrificed by cervical dislocation at 4 hours after MAA treatment. The forelimb buds were isolated from both control and treated group embryos and were then homogenized. The crude extracts were then fractionated with ammonium sulfate and each fraction was analyzed by 1-D and 2-D SDS-PAGE techniques respectively. The 1-D and 2-D electrophoregrams revealed that in the treated group of protein fraction 20-40% ammonium sulfate (F-II), a protein of 31.0-36.5 kDa and a protein spot 35.1 kDa, pI 6.2 could be detected, which was not found in the control. In the treated group of protein fraction 40-60% ammonium sulfate (F-III) a protein of 66.3-97.4 kDa and a protein spot 81.7 kDa, pl 7.3 could be detected which was not found in the control, whereas in the control group a protein of 36.5-55.4 kDa, which is a protein spot 41.6 and pI 6.4, was detected but not detected in the treated group. It could be concluded from this experiment hat in the mouse forelimb buds, MAA treatment induce the protein expression of two proteins (35.1 kDa, pI 6.2 and 81.7 kDa, 7.3) and inhibit the expression of one protein (41.6p, pI 6.4).

Keywords: mouse embryo; limb development; methoxyacetic acid; protein profile

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