Djati, M. Sasmito and Wahjuningsih, Sri (2006) Perubahan Struktur Sitoskeleton Berbasis Mikrotubulus dan Ultrastruktur Oosit Pasca Kriopreservasi Dengan Metode Vitrifikasi. Project Report. Fakultas Peternakan Universitas Brawijaya, Malang. (Unpublished)
Abstract
Tujuan Penelitian tahun pertama adalah : mengkaji kualitas oosit pasca vitrifikasi terhadap morfologi, viabilitas, serta struktur sitoskeleton oosit berbasis mikrotubulus dan mempelajari faktor yang menyebabkan terjadinya penurunan kualitas oosit pasca vitrifikasi melalui evaluasi morfologi, viabilitas serta analisa mikrotubulus oosit menggunakan metode imunohistokimia. Materi penelitian adalah oosit kambing yang telah dilakukan maturasi in vitro selama 24 jam. Rancangan percobaan adalah : (1) rancangan acak lengkap pola faktorial 6x3, dengan perlakuan konsentrasi EG 0, 10, 20, 30, 40, dan 50 % dan lama paparan 1,3,5 menit untuk evaluasi morfologi dan viabilitas dan (2) deskriptif untuk analisa struktur sitoskeleton. Hasil penelitian menunjukkan bahwa konsentrasi EG dan lama paparan berpengaruh terhadap morfologi dan viabilitas (p<0.01). Persentase oosit dengan morfologi normal dan oosit hidup tertinggi serta vitrifikasi menggunakan 30 % EG dan lama paparan 3 menit, masing-masing sebesar 86.6% dan 78.7 %. Konsentrasi EG 30 % dan lama paparan 3 menit merupakan perlakuan terbaik untuk melindungi oosit pada proses vitrifikasi. Hasil pengamatan terhadap sitoskeleton berbasis mikrotubulus menggunakan metode imunohistokimia diperoleh gambar yang menunjukkan adanya perbedaan struktur sitoskeleton oosit kontrol dan oosit hasil vitrifikasi. Pada perlakuan kontrol struktur sitoskeleton berbasis mikrotubul tampak seperti benang-benang berwarna kecoklatan. Hasil penelitian menunjukkan bahwa setelah proses vitrifikasi struktur sitoskeleton mengalami perubahan. Kesimpulan dari penelitian ini adalah vitrifikasi menggunakan 30 % EG dan lama paparan 3 menit dapat menghambat kerusakan morfologi oosit dan penurunan viabilitas yang disebabkan oleh proses vitrifikasi. Untuk menjaga fungsi oosit, struktur sitoskeleton berbasis mikrotubulus harus dapat dipertahankan. EG sebesar 30 % terbukti dapat mencegah kerusakan lebih lanjut dari sitoskeleton. Struktur sitoskeleton pada perlakuan 30 % EG tampak seperti benang yang berarti masih bisa mengalami recovery, sedangkan pada konsentrasi EG yang lain, sitoskeleton oosit tampak mengalami kerusakan. Terdapat hubungan antara morfologi, viabilitas oosit dan struktur sitoskeleton berbasis mikrotubulus. Disarankan melakukan analisa ultrastruktur oosit menggunakan Transmission Electron Microscopy agar dapat menjawab mengapa terjadi penurunan kualitas oosit pasca vitrifikasi.
English Abstract
The aim of this research for first year was to : study the quality of oocyte after vitrification to morphology, viability and also oocyte cytoskeleton structure based on microtubulus and study the factor that causing the happening of oocyte quality degradation after vitrification by morphology evaluation, viability and also analyse oocyte microtubulus structure after vitrification using imunohistochemistry method. The material of this research was used goat oocyte which have been maturated by in vitro during 24 hours. The research use (1) random complete design with factorial pattern 6x3, with treatment concentration of Ethylene Glycol (EG) were 0, 10, 20, 30, 40 and 50 % with the time of exposure during 1,3,5 minutes for the morphology and viability evaluation and (2) descriptive for the structure of cytoskeleton. The results of the research showed that concentration of EG and the time of exposure influence morphology and viability (p<0.01). Percentage of oocyte with normal morphology and highest life oocyte and also vitrification use 30% EG and the time of exposure during 3 minutes, each of 86.6% and 78.7%. Concentration of EG 30% and the time of exposure during 3 minutes were the best treatment to protect oocyte in process of vitrification. The result of observation to cytosceleton based on microtubulus use imunohistochemistry method was obtained picture that showing the difference existence of cytosceleton structure of oocyte control and cryopreserved oocyte. In the control treatment, the structure of cytosceleton based on microtubulus looked like brown yarn. The result of this research showed that after the process of vitrification, structure of cytoskeleton has a change. The conclusion of this research was that vitrification use 30% EG and the time of exposure during 3 minutes can pursue damage of oocyte morphology and viability which is caused by the process of vitrification. To safe the function of oocyte, cytosceleton structure based on microtubulus have to earn to be defended. EG equal to 30% proven can prevent further damage of cytosceleton. Cytosceleton structure in the treatment of 30% EG looked like yarn means that it still can recovery, while in the other concentration of EG, visible oocyte cyitosceleton experience of damage. There are relation between morphology, oocyte viability and cytosceleton structure based on microtubulus.
| Item Type: | Monograph (Project Report) |
|---|---|
| Identification Number: | PEN/571.654/WAH/p/2006/020603803 |
| Subjects: | 500 Natural sciences and mathematics > 571 Physiology and related subjects > 571.6 Cell biology > 571.65 Cytoplasm > 571.654 Cytoskeleton |
| Divisions: | Fakultas Peternakan > Peternakan |
| Depositing User: | soegeng sugeng |
| Date Deposited: | 13 Sep 2018 06:48 |
| Last Modified: | 16 Jun 2022 07:44 |
| URI: | http://repository.ub.ac.id/id/eprint/11964 |
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