Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/90277
標題: Maturation and development of porcine and canine oocytes after Indirubin treatment
Indirubin對豬與犬卵母細胞體外成熟與發育能力之影響
作者: Sing-Ping Chuang
莊欣萍
關鍵字: 成熟促進因子
卵丘卵母細胞複合體
maturation promoting factor,MPF
cumulus-oocyte complexes, COCs
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摘要: Indirubin-3'-oxime(Indirubin)為一CDK轉磷酸酶抑制劑,主要是藉由與ATP競爭p34cdc2 kinases之ATP結合位置,進而阻斷其磷酸化作用,導致成熟促進因子(maturation promoting factor, MPF)無法致活,而抑制卵母細胞原泡核(germinal vesicle, GV)的崩解(GV breakdown, GVBD),此一作用有利於原泡核轉置操作並改善卵母細胞後續發育能力。本試驗之目的為探討Indirubin對豬卵母細胞體外成熟與其孤雌胚體外發育之影響。試驗一,豬卵丘卵母細胞複合體(cumulus-oocyte complexes, COCs)培養於含不同濃度Indirubin (0, 1.56, 3.13, 6.25與12.5 μM)之NCSU-23培養液中,經22 h後轉至不含Indirubin之NCSU-23中繼續培養44 h,以觀察COCs之擴散與ERK 1/2磷酸化程度。結果顯示,豬卵母細胞經過Indirubin處理22 h後,COCs外圍卵丘細胞擴散程度隨Indirubin濃度之增加而減少,而在較高濃度Indirubin(6.25與12.5 μM)處理下COCs無擴散現象,ERK1/2之磷酸化程度亦與GV期之卵母細胞相似,但低於對照組者(P < 0.05)。試驗二,卵母細胞成熟後經孤雌激活後再觀察其發育情形。經移除Indirubin並延長培養44 h後,豬卵母細胞成熟率於各處理組(90.4-91.4 %)、對照組(44 h;90.4 %)及延長培養組(66 h,89.3 %)之間並無差異(P > 0.05);成熟卵母細胞經激活(2.2 kV/cm, 30 μsec與2.5 mM 6-DMAP, 4 h)後,各組間之分裂率亦相近(88.6-95.8 %),但囊胚率在延長培養之對照組(17.4 %)與Indirubin各處理組(20.7-28.7 %)顯著低於對照組(51.7 %, P < 0.05)。試驗三,取自不同月齡家犬的卵巢之卵母細胞,以Indirubin處理後與未處理組繼續培養72 h,卵母細胞皆無法恢復減數分裂(< 10 %)。以上結果顯示適當濃度之Indirubin可有效抑制豬卵母細胞停留於GV期,但移除抑制劑後其囊胚率降低,可能與延長體外培養時間或殘留於細胞內之濃度有關;另外,犬卵母細胞經Indirubin處理並未改善其成熟率,此仍需更多之研究。
URI: http://hdl.handle.net/11455/90277
文章公開時間: 2014-01-28
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