Protein yang Terkait dengan Teratogenisitas Anggota Tubuh Mencit Swiss Webster Akibat Perlakuan dengan Asam Metoksiasetat (MAA)
Keywords:
embrio mencit, perkembangan anggota tubuh, asam metoksiasetat, profil protein, mouse embryo, limb development, methoxyacetic acid, protein profile.Abstract
Telah diteliti protein yang terkait dengan teratogenesis anggota tubuh mencit Swiss Webster akibat perlakuan dengan MAA. Mencit umur kebuntingan 11 hari diberi perlakuan dosis tunggal MAA 10 mmol/kg berat badan secara gavage, sedangkan kelompok kontrol hanya diberi pelarut akuabides steril. Mencit bunting dibunuh secara dislokasi leher 4 jam setelah perlakuan dengan MAA. Tunas anggota tubuh depan diisolasi dari kelompok kontrol dan perlakuan lalu dihomogenisasi. Ekstrak kasar kemudian difraksinasi dengan amonium sulfat dan masing-masing fraksi dianalisis dengan teknik l-D dan 2-D SDS-PAGE. Elektroforegram l-D dan 2-D menunjukkan bahwa pada kelompok perlakuan fraksi protein ammonium sulfat 20-40 % (F-lI), dapat dideteksi protein 31,0-36,5 kDa serta bercak protein 35,1 kDa, pI 6,2 yang tidak terdapat pada kontrol. Pada kelompok perlakuan fraksi protein ammonium sulfat 40-60% (F-lll), dapat dideteksi protein 66,3-97,4 kDa serta bercak protein 8 I,7 kDa, pl 7,3 yang tidak terdapat pada kontrol. Sedangkan pada kelompok kontrol F-llI, protein 36,5-55,4 kDa sertab ercakp rotein 41,6 kDa, pI 6,4 terdeteksi, tetapi tidak terdeteksi pada kelompok perlakuan. Dari penelitian ini dapat disimpulkan bahwa pada tunas anggota tubuh depan mencit, perlakuan dengan MAA menginduksi ekspresi dua protein(35,1 kDa, pl 6,2 dan 81,7 kDa, pl 7,3) dan menghambat ekspresi satu protein(41,6 kDa, pl 6,4).
Proteins which are Linked with Swiss Webster Mouse Limb Teratogenesis as the Effects of Methoxyacetic Acid (MAA) Treatment
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 l-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, pl 6.2 could be detected, which was not found in the control. In the treated group of protein fraction 40-60% ammonium sulfate( F-lll) 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 spot4 1.6 and p l 6.4, was detected but not detected in the treated group.It could be concluded from this experiment that in the mouse forelimb buds, MAA treatment induce the protein expression of two proteins(35.1 kDa, pl 6.2 and 81.7 kDa, 7.3) and inhibit the expression of one protein (41.6p, l 6.4).
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