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白血病抑制因子及其受体在人胎儿和成人卵巢中的免疫细胞化学检测及逆转录聚合酶链反应表达

Immunocytochemical detection and RT-PCR expression of leukaemia inhibitory factor and its receptor in human fetal and adult ovaries.

作者信息

Abir R, Fisch B, Jin S, Barnnet M, Freimann S, Van den Hurk R, Feldberg D, Nitke S, Krissi H, Ao A

机构信息

Infertility and IVF Unit, Department of Obstetrics and Gynecology, Rabin Medical Center, Beilinson Campus, Petah Tikva 49100 Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.

出版信息

Mol Hum Reprod. 2004 May;10(5):313-9. doi: 10.1093/molehr/gah047. Epub 2004 Mar 25.

Abstract

The ability to mature human primordial follicles in vitro would assist fertility restoration. However, the signals initiating growth of primordial follicles are unknown. Growth factors such as leukaemia inhibitory factor (LIF) may play a role in this process. To investigate the expression of LIF and its receptor in early developing follicles, nine ovarian samples from adolescents/adults aged 13-43 years and 23 ovaries from human fetuses aged 19-33 gestational weeks were immediately fixed or frozen. The fixed samples were prepared for a study of immunocytochemical staining of LIF and its two receptor units (LIF-R and gp 130). mRNA was extracted from the frozen ovarian samples, and the expression of LIF, LIF-R and gp 130 was investigated by RT-PCR. Products were resolved by 10% polyacrylamide gel electrophoresis and image analysis. There was strong to moderate immunocytochemical staining for LIF and LIF-R in oocytes from the primordial follicular stages onwards, and very weak to moderate staining for gp 130. LIF-R was also detected in granulosa cells of primary and secondary follicles from adolescents/adults. Transcripts of LIF, LIF-R and gp 130 RNA were identified by RT-PCR in all samples. The immunocytochemical staining and mRNA expression of LIF and its receptor are consistent with the concept that LIF might be involved in growth initiation of human primordial follicles through its receptor.

摘要

在体外使人类原始卵泡成熟的能力将有助于恢复生育能力。然而,启动原始卵泡生长的信号尚不清楚。白血病抑制因子(LIF)等生长因子可能在此过程中发挥作用。为了研究LIF及其受体在早期发育卵泡中的表达,对9份来自13 - 43岁青少年/成年人的卵巢样本以及23份来自妊娠19 - 33周人类胎儿的卵巢立即进行固定或冷冻。对固定样本进行LIF及其两个受体亚基(LIF-R和gp130)的免疫细胞化学染色研究。从冷冻的卵巢样本中提取mRNA,并通过逆转录聚合酶链反应(RT-PCR)研究LIF、LIF-R和gp130的表达。产物通过10%聚丙烯酰胺凝胶电泳和图像分析进行分离。从原始卵泡阶段开始,卵母细胞中LIF和LIF-R有强至中度免疫细胞化学染色,而gp130有非常弱至中度染色。在青少年/成年人初级和次级卵泡的颗粒细胞中也检测到LIF-R。通过RT-PCR在所有样本中鉴定出LIF、LIF-R和gp130 RNA的转录本。LIF及其受体的免疫细胞化学染色和mRNA表达与LIF可能通过其受体参与人类原始卵泡生长启动的概念一致。

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