IN-SITU LOCALIZATION OF APOPTOSIS IN THE RAT OVARY DURING FOLLICULAR ATRESIA
IN-SITU LOCALIZATION OF APOPTOSIS IN THE RAT OVARY DURING FOLLICULAR ATRESIA
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DOI：
10.1095/biolreprod51.5.888
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发表时间：
1994-11-01
影响因子：
3.6
通讯作者：
YEH, J
中科院分区：
文献类型：
--
作者：
PALUMBO, A;YEH, J
细胞凋亡是发生在许多组织中的一种生理性细胞死亡，并且可以由肽生长因子调节。最近的研究表明，在包括大鼠、猪、鸡、狒狒和兔在内的几种动物的卵泡闭锁期间，卵巢中发生了细胞凋亡。本研究的目的是证明，通过在原位鉴定的凋亡细胞在完整的卵巢切片，在不同的功能状态下，在大鼠卵巢细胞凋亡发生的网站。我们评估了三种模型中细胞凋亡的存在：未成年大鼠，ECG治疗的大鼠和成年自行车大鼠。用蛋白酶K预处理石蜡卵巢切片，然后通过与酶末端脱氧核苷酸转移酶（TDT）孵育，用生物素化脱氧尿苷三磷酸（dUTP）进行末端标记。然后通过使用抗生物素蛋白缀合的过氧化物酶以3，3 '-二氨基联苯胺作为底物对其进行染色。健康的窦状卵泡和腔前卵泡无染色。窦前卵泡和窦闭锁卵泡颗粒细胞核均呈阳性染色。散在的卵泡膜细胞也被染色。基质细胞始终呈阴性。阳性对照是用DNA酶I预处理的切片;这些显示所有细胞核的强染色。阴性对照，其中末端TDT或其生物素化底物被省略，是适当的阴性。本研究系统地分析了不同功能阶段大鼠卵巢细胞凋亡的情况，支持了细胞凋亡参与卵泡闭锁过程的假说
Apoptosis is a type of physiologic cell death that occurs in many tissues and may be regulated by peptide growth factors. Recent studies indicate that apoptosis occurs in the ovary during follicular atresia in several animal species, including the rat, pig, chicken, baboon, and rabbit. The purpose of this study was to demonstrate, through in situ identification of apoptotic cells in intact ovarian sections, the sites in which apoptosis occurs in the rat ovary in different functional states. We evaluated the presence of apoptosis in three models: immature rats, eCG-treated rats and adult cycling rats. Paraffin ovarian sections were pretreated with proteinase K and then end-labeled with biotinylated deoxyuridine triphosphate (dUTP) by incubation with the enzyme terminal deoxynucleotidyl transferase (TDT). They were then stained through use of avidin-conjugated peroxidase with 3,3'-diaminobenzidine as the substrate. Healthy antral and preantral follicles had no staining. The nuclei of granulosa cells of preantral and antral atretic follicles were positively stained in all the animal groups. Scattered theca cells were also stained. Stromal cells were consistently negative. Positive controls were sections pretreated with DNase I; these displayed intense staining of all nuclei. Negative controls, in which either terminal TDT or its biotinylated substrate was omitted, were appropriately negative. This study represents a systematic analysis of apoptosis in the rat ovary at different functional stages and supports the hypothesis that apoptosis is involved in the process of follicular atresia