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소 난구복합체의 체외성숙시 Okadaic Acid (OA)가 핵성숙 및 Mitochondria 활성에 미치는 영향 KCI 등재
Effect of Okadaic Acids (OA) on Nuclear Maturation and Mitochondrial Activity of Hanwoo COCs during in vitro Maturation
pp.303-309
본 연구는 한우 난구 복합체의 체외성숙에서 OA가 미치는 영향에 대하여 조사하였다. 도축 한우암소의 난소로부터 난구 복합체를 채취하여 PVA-TCM199로 3회 이상 세정 후 PVA-TCM 199, 0.2 uM, 2 uM, 20 uM OA를 각각 첨가하여 공기, 에서 6, 12, 24시간 동안 체외성숙을 실시하였다. 또한 체외성숙시 cycloheximide(CX)와 OA와 체외성숙 효과를 확인하기 위하여 0.1M-PVA TCM199, CX 25 ug/m
This study was conducted to examine the effects of OA on metaphase of meiosis II and the mitochondrial activity of cytoplasm in bovine cumulus oocytes complexes(COCs) during in vitro maturation. Hanwoo COCs were collected from the slaughterhouse cow ovaries and matured in TCM199 supplemented with PVA, 0.2 uM, 2 uM, 20 uM OA for the maturation rate of OA concentration. For the maturation effects between OA and cycloheximide(CX), COCs were matured in TCM199 with 25 ug/mL CX, 25 ug/mL CX (6 hrs culture) plus 2 uM OA or 2 uM OA only at a atmosphere air for 6, 12, 24 hrs. To evaluate the nuclear types of matured COCs, cumulus cells were removedby hyaluronidase sol. and oocytes were fixed in 1:3 acetic acid ethyl alcohol for 30 sec. and then stained with basic Fuchsin sol. For the detection of fluoriscent intensity (FI) of matures oocytes, cumulus cells were removed same as performed above and were stained with 20 nM mite tracker for 20 min. at . Mitochondrial activity of FI in matured oocytes was imaged by laser conforcal microscopy (Fluoview, Olympus, Japan) and were measured scanned face on 5 um from median to endpoint of oocytes. Statical analysis of nuclear types observed the three replicates was carried out with ANOVA and Fisher's protected least significant difference test using the STATVIEW program. FI of matures oocytes was compared the multiples of the least intensity among the measured oocytes. Maturing in TCM199 supplemented with PVA, 0.2 uM, 2 uM, 20 uM OA, metaphase B were showed 72.0, 50.0, 70.0, , respectively and there were different significant(p<0.05). In the case of treatment with OA and CX, metaphase were in PVA-TCM199, 25 ug/mL CX, 25 ug/mL CX plus OA or 2uM OA only, respeclively. FI was revealed the increasing tendency during the process of maturation. Whereas FI in CX was decreased about 3 times compared to the other treatments of 6 hrs maturation. We conclude that OA regulates bovine COCs maturation and induces the mitochondrial activity during the process of maturation.
