Internucleosomal DNA fragmentation in ovine luteal tissue associated with luteolysis: in vivo and in vitro analyses.
Internucleosomal DNA fragmentation in ovine luteal tissue associated with luteolysis: in vivo and in vitro analyses.
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DOI：
10.1095/biolreprod52.2.305
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发表时间：
1995-02
影响因子：
3.6
通讯作者：
Bo R. Rueda;Julie A. Wegner;S. Marion;Derron D. Wahlen;Patricia B. Hoyer
中科院分区：
文献类型：
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作者：
Bo R. Rueda;Julie A. Wegner;S. Marion;Derron D. Wahlen;Patricia B. Hoyer
核小体间 DNA 碎片是细胞凋亡的一个特征，可以通过琼脂糖凝胶电泳将其可视化为离散的低分子量 DNA 片段（梯状），大约为 185 bp 的倍数。 CL 是从超排卵母羊（对照）或在注射 hCG 后不同天注射前列腺素 F2 α（PGF2 α）后 12 小时收集的。评估了 PGF2 α 在第 8、10、12 和 14 天（每次治疗每天 n > 或 = 3 次）诱导黄体细胞 DNA 片段化的能力。分离 DNA 并在琼脂糖凝胶上观察。第 8、10 或 12 天的对照母羊的 CL 中未观察到 DNA 断裂。在第 14 天对照母羊的 CL 中，四只动物中的两只观察到 DNA 核小体间断裂（指示细胞凋亡）以及非特异性 DNA 断裂（指示非凋亡细胞死亡）。此外，在全天接受 PGF2 α 处理的母羊的 CL 中观察到这种 DNA 片段化模式。解离后在黄体组织中观察到 DNA 片段化的证据，但在从第 10 天对照母羊（孵育 4 小时）收集的未切片、未解离 CL 或切片、未孵育 CL 中未观察到片段化。仅切片和孵育就足以引发 DNA 片段化。人们采用了多种方法来抑制 DNA 片段化。在整个 4 小时的孵育期间，只有在孵育介质中添加醋酸锌 (1 mM) 才能防止切片引发的 DNA 片段化 (p < 0.05)。因此，似乎存在一种或多种黄体内因子，可直接启动与黄体溶解过程中的细胞死亡相关的 DNA 断裂。（摘要截断为 250 字）
Internucleosomal DNA fragmentation, a characteristic of apoptosis, can be visualized with agarose gel electrophoresis as discrete low-molecular-weight DNA fragments (laddering), in multiples of approximately 185 bp. CL were collected from superovulated ewes (control) or at 12 h after injection of prostaglandin F2 alpha (PGF2 alpha) on various days after hCG injection. The ability of PGF2 alpha on Days 8, 10, 12, and 14 (n > or = 3 per day per treatment) to induce luteal cell DNA fragmentation was evaluated. DNA was isolated and visualized on agarose gels. No DNA fragmentation was observed in CL from control ewes on Days 8, 10, or 12. Internucleosomal fragmentation of DNA (indicative of apoptosis) as well as nonspecific DNA fragmentation (indicative of non-apoptotic cell death) in CL from Day 14 controls was observed in two of four animals. Additionally, this pattern of DNA fragmentation was observed in CL from ewes treated with PGF2 alpha on all days. Evidence of DNA fragmentation was observed in luteal tissue after dissociation, yet no fragmentation was observed in unsliced, non-dissociated CL collected from Day 10 control ewes (incubated 4 h), or in sliced, non-incubated CL. Slicing and incubation alone were sufficient to initiate DNA fragmentation. A variety of approaches were utilized to inhibit DNA fragmentation. Only the addition of zinc acetate (1 mM) in the incubation medium throughout the 4-h incubation period prevented DNA fragmentation that was initiated by slicing (p < 0.05). There appear therefore, to be one or more intraluteal factors that directly initiate DNA fragmentation associated with cell death in luteolysis.(ABSTRACT TRUNCATED AT 250 WORDS)