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小鼠植入前胚胎中转化生长因子β1(TGFβ1)mRNA表达的定量分析以及小鼠胚胎和生殖道中TGFβ受体(I型和II型)表达的测定。

Quantification of transforming growth factor beta1 (TGFbeta1) mRNA expression in mouse preimplantation embryos and determination of TGFbeta receptor (type I and type II) expression in mouse embryos and reproductive tract.

作者信息

Chow J F, Lee K F, Chan S T, Yeung W S

机构信息

Department of Obstetrics and Gynaecology, The University of Hong Kong, Queen Mary Hospital, Pokfulam Road, Hong Kong, China.

出版信息

Mol Hum Reprod. 2001 Nov;7(11):1047-56. doi: 10.1093/molehr/7.11.1047.

Abstract

We hypothesized that transforming growth factor beta1 (TGFbeta(1)) and its receptors play a role in the interaction between the preimplantation embryo and the reproductive tract. To investigate this hypothesis, TGFbeta 1 mRNA in mouse embryos was quantified by competitive reverse transcription-polymerase chain reaction using an RNA mimic. TGFbeta 1 was first detected in the unfertilized oocyte, disappeared after fertilization and was expressed again at the 2-cell stage (4410 +/- 1330 transcripts/embryo). Its expression increased gradually, peaked at the 8-cell stage (58 600 +/- 17 300 transcripts/embryo) and declined rapidly after the morula stage reaching a concentration of 1520 +/- 546 transcripts/embryo at the blastocyst stage. The mRNA levels of TGFbeta 1 at the 8-cell and morula stages were significantly higher than that at other cell stages (P < 0.05). The expression of TGF receptors in embryos and in the reproductive tract was also investigated. Both TGFbeta(1) type I (ALK-5) and type II TGFbeta receptors were detected in embryos from 1-cell to blastocyst stage by immunohistochemistry. Northern hybridization and immunohistochemistry showed a constant expression of both TGFbeta receptors in the oviduct from day 1 to day 4 of pregnancy, whilst in the uterus there was a marked increase in the expression of TGFbeta type I receptor on day 3. Expression of TGFbeta type II receptor in the uterus remained unaltered throughout the study period. This study has shown that preimplantation mouse embryos produce TGFbeta(1) and that both the embryos and the reproductive tract are responsive to TGFbeta(1) in the preimplantation period.

摘要

我们推测,转化生长因子β1(TGFβ(1))及其受体在着床前胚胎与生殖道之间的相互作用中发挥作用。为了验证这一推测,我们使用RNA模拟物通过竞争性逆转录 - 聚合酶链反应对小鼠胚胎中的TGFβ1 mRNA进行了定量分析。TGFβ1最初在未受精卵母细胞中被检测到,受精后消失,并在2细胞期再次表达(4410±1330转录本/胚胎)。其表达逐渐增加,在8细胞期达到峰值(58600±17300转录本/胚胎),桑椹胚期后迅速下降,在囊胚期降至1520±546转录本/胚胎的浓度。8细胞期和桑椹胚期的TGFβ1 mRNA水平显著高于其他细胞期(P < 0.05)。我们还研究了胚胎和生殖道中TGF受体的表达。通过免疫组织化学在1细胞期至囊胚期的胚胎中均检测到了TGFβ(1) I型(ALK - 5)和II型TGFβ受体。Northern杂交和免疫组织化学显示,在妊娠第1天至第4天,输卵管中两种TGFβ受体的表达持续稳定,而在子宫中,TGFβ I型受体的表达在第3天显著增加。在整个研究期间,子宫中TGFβ II型受体的表达保持不变。这项研究表明,着床前小鼠胚胎可产生TGFβ(1),并且在着床前期胚胎和生殖道均对TGFβ(1)有反应。

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