Expression of vimetin protein and neurofilamen on forelimb buds of black-6 mice on gestation day 12 induced by 2-methoxyethanol by RTPCR

YULIA IRNIDAYANTI

Abstract. Irnidayanti Y. 2010. Expression of vimetin protein and neurofilamen on forelimb buds of black-6 mice on gestation day 12 induced by 2-methoxyethanol by Real Time RT-PCR. Nusantara Bioscience 2: 116-120. The aim of this study was to investigate impact
of 2-methoxyethanol, a major industrial chemical of plastic. Gene expression analysis is increasingly important in biological research, while real-time reverse transcription PCR (RT-PCR) is becoming the method of choice for high-through put and accurate expression
profiling of selected genes. Pregnant black-6 mice were injected intraperitoneally to 7.5 mmol/kg of 2- methoxyethanol on gestation day (GD) 10. Embryo were obtained on gestation day 12. Forelimb buds of embryo was collected and then put in the tube, which containing RNA-latter solution. To identify gene expression changes in forelimb bud caused induction 2-methoxyethanol, Real Time PCR were using in this research. For the experiments the real-time RT-PCR Light Cycler technology was used. The results suggested that injection of 2- methoxyethanol, in prenatal period especially on gestation day 12, the expression of vimentin in forelimb buds of mice treatment increase than control mice. Meanwhile the expression of neurofilament tended to decrease, indirectly is not caused by the injection of 2- methoxyethanol.
Key words: vimentin, neurofilament, 2-methoxyethanol, limb bud, black-6 mice.

Abstrak. Irnidayanti Y. 2010. Ekspresi protein vimetin dan neurofilamen pada tunas anggota depan mencit black-6 umur kebuntingan 12 hari akibat induksi 2-metoksietanol secara Real Time RT-PCR. Nusantara Bioscience 2: 116-120. Tujuan penelitian ini adalah untuk mengetahui dampak dari 2-metoksietanol, bahan kimia utama industri plastik. Analisis ekspresi gen semakin penting dalam penelitian biologi, dimana real-time reverse transcription PCR (RT-PCR) menjadi metode yang dipilih untuk mendapatkan profil ekspresi dari seleksi gen secara akurat. Mencit black-6 bunting diinjeksi 2-metoksietanol secara intraperitoneal dengan 7,5 mmol/kg pada umur kebuntingan 10 hari. Embrio diperoleh pada umur kebuntingan 12 hari. Tunas anggota depan embrio dikumpulkan dan dimasukkan ke dalam tabung, yang mengandung larutan RNA-latter. Untuk mengetahui perubahan ekspresi gen tunas anggota depan yang disebabkan oleh induksi 2-metoksietanol digunakan RT-PCR dalam penelitian ini. Dalam percobaan digunakan teknologi RT-PCR Light Cycler. Hasil penelitian menunjukkan bahwa injeksi 2- metoksietanol, pada periode pralahir terutama pada umur kebuntingan 12 hari, ekspresi vimentin pada tunas anggota depan mencit perlakuan meningkatkan dibandingkan kontrol. Sementara itu ekspresi neurofilamencenderung menurun, secara tidak langsung tidak disebabkan oleh injeksi 2- metoksietanol.
Kata kunci: vimentin, neurofilamen, 2-metoksietanol, limb bud, mencit black-6.

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