Caspase-3 Is a Pivotal Mediator of Apoptosis during Regression of the Ovarian Corpus Luteum.
Caspase-3 Is a Pivotal Mediator of Apoptosis during Regression of the Ovarian Corpus Luteum.
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DOI：
10.1210/endo.143.4.8726
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发表时间：
2002-04
期刊：
影响因子：
4.8
通讯作者：
S. Carambula;T. Matikainen;M. P. Lynch;R. Flavell;Paulo B Dias Gonc Alves;J. Tilly;B. Rueda
中科院分区：
文献类型：
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作者：
S. Carambula;T. Matikainen;M. P. Lynch;R. Flavell;Paulo B Dias Gonc Alves;J. Tilly;B. Rueda
由于caspase-3被认为是细胞凋亡的主要刽子手，并被认为是黄体退化的中介，我们假设caspase-3缺失小鼠的黄体（CL）在黄体退化过程中表现出延迟的细胞凋亡，与野生型（WT）小鼠的相比。为了验证这一假设，在未成熟（产后24 -27）WT和caspase-3缺陷的雌胎中，外源性促性腺激素可以同步排卵。在排卵后30小时，通过手工解剖分离个体CL，并置于无血清和生长因子的器官培养皿中。在分离（0小时）和培养24、48和72小时后，取出细胞，通过多种标准评估加工（活性）caspase-3酶的存在和细胞凋亡情况。在培养前，WT或caspase-3缺失的CL中没有活性caspase-3酶或凋亡的证据。然而，在培养过程中，来自WT小鼠的CL表现出活性caspase-3水平和凋亡水平的时间依赖性增加。相比之下，平行培养的caspase-3缺陷小鼠的CL没有表现出任何可检测到的活性caspase-3，并且凋亡率降低。为了扩展体外培养实验的结果，我们在排卵后2、4、6天收集WT和caspase-3缺失的雌性窝鼠卵巢，分析细胞凋亡的发生情况。WT小鼠的卵巢在排卵后第6天只有残留的黄体组织，而caspase-3缺陷小鼠的卵巢在排卵后第6天保留了许多CL，其大小与WT小鼠早期黄体期观察到的相似。重要的是，在排卵后的任何时间点检测caspase-3缺陷小鼠的CL中，细胞凋亡都没有显著增加，这表明复归过程确实被推迟了。相比之下，无论基因型如何，黄体酮水平都有所下降。这些数据提供了第一个直接证据，证明caspase-3在功能上是黄体退化期间细胞凋亡正常进行所必需的。然而，caspase-3并不是与黄体溶解相关的类固醇生成减少的直接中介。（内分泌学143:1495-1501,2002）
Because caspase-3 is considered a primary executioner of apoptosis and has been implicated as a mediator of luteal regression, we hypothesized that corpora lutea (CL) derived from caspase-3 null mice would exhibit a delayed onset of apoptosis during luteal regression, when compared with CL derived from wild-type (WT) mice. To test this hypothesis, ovulation was synchronized in immature (postpartum d 24 –27) WT and caspase-3-deficient female littermates by exogenous gonadotropins. Individual CL were isolated by manual dissection, 30 h after ovulation, and placed in organ culture dishes in the absence of serum and growth factors. At the time of isolation (0 h) and after 24, 48, and 72 h in culture, the CL were removed and assessed for the presence of processed (active) caspase-3 enzyme and for apoptosis by multiple criteria. There was no evidence of active caspase-3 enzyme or apoptosis in either WT or caspase-3-deficient CL before culture. However, CL derived from the WT mice exhibited a time-dependent increase in the level of active caspase-3 and apoptosis during culture. By comparison, CL derived from caspase-3-deficient mice, cultured in parallel, failed to exhibit any detectable active caspase-3 and showed attenuated rates of apoptosis. To extend these findings derived from ex vivo culture experiments, ovaries were collected from WT and caspase-3 null female littermates at 2, 4, or 6 d post ovulation, and the occurrence of apoptosis within the CL was analyzed. Whereas ovaries of WT mice had only residual luteal tissue at d 6 post ovulation, ovaries collected from caspase-3-deficient mice retained many CL, at d 6 post ovulation, that were similar in size to those observed in the early luteal phase of WT mice. Importantly, there was no dramatic increase in apoptosis in CL of caspase-3-deficient mice at any time point examined post ovulation, indicating that the involution process had indeed been delayed. In contrast, the levels of progesterone declined regardless of genotype. These data provide the first direct evidence that caspase-3 is functionally required for apoptosis to proceed normally during luteal regression. However, caspase-3 is not a direct mediator of the decrease in steroidogenesis associated with luteolysis. (Endocrinology 143: 1495–1501, 2002)