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表皮生长因子及其受体基因在猪卵巢卵泡中的表达及肽定位

Epidermal growth factor and its receptor gene expression and peptide localization in porcine ovarian follicles.

作者信息

Singh B, Rutledge J M, Armstrong D T

机构信息

Department of Obstetrics and Gynaecology, University of Western Ontario, London, Canada.

出版信息

Mol Reprod Dev. 1995 Apr;40(4):391-9. doi: 10.1002/mrd.1080400402.

Abstract

The present study was undertaken to determine the expression of genes for epidermal growth factor (EGF) and its receptor (EGF-R) in various components of medium-sized porcine ovarian follicles by reverse transcription-polymerase chain reaction (RT-PCR), and to localize their peptides during folliculogenesis by immunocytochemistry. A strong band for EGF mRNA transcript was detected in the oocyte, whereas the signal in cumulus, granulosa, and theca cells was very weak but detectable. In contrast, a very strong EGF-R mRNA signal was observed in cumulus, granulosa, and theca cells, whereas the signal in the oocyte was very weak. EGF peptide was localized in the oocyte, cumulus, and granulosa cells of all stages of follicle. In the oocyte, the intensity of immunostaining was more pronounced in primordial and primary follicles, compared to atrial follicles. In large antral follicles, immunostaining was pronounced in granulosa cells, whereas theca cells showed little or no detectable staining for EGF. EGF staining was also observed in the cumulus and granulosa cells of follicles undergoing atresia. EGF-R immunostaining was observed in the oocytes of primordial and primary follicles, and in cumulus, granulosa, and theca cells of all stages of follicle, including atretic follicles. In large antral follicles, the intensity of immunostaining was more pronounced in theca cells than in granulosa cells, and the oocyte showed little or no detectable staining. No immunostaining was observed when the primary antibody was replaced with preimmune serum (EGF), or preabsorbed with the control peptide (EGF-R), confirming the specificity of the staining procedures.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

本研究旨在通过逆转录-聚合酶链反应(RT-PCR)确定表皮生长因子(EGF)及其受体(EGF-R)基因在中等大小猪卵巢卵泡各组分中的表达,并通过免疫细胞化学在卵泡发生过程中定位其肽段。在卵母细胞中检测到一条强的EGF mRNA转录带,而在卵丘、颗粒细胞和卵泡膜细胞中的信号非常弱但可检测到。相反,在卵丘、颗粒细胞和卵泡膜细胞中观察到非常强的EGF-R mRNA信号,而卵母细胞中的信号非常弱。EGF肽定位于卵泡各阶段的卵母细胞、卵丘和颗粒细胞中。在卵母细胞中,与有腔卵泡相比,原始卵泡和初级卵泡中的免疫染色强度更明显。在大型有腔卵泡中,颗粒细胞中免疫染色明显,而卵泡膜细胞对EGF的染色很少或未检测到。在闭锁卵泡的卵丘和颗粒细胞中也观察到EGF染色。在原始卵泡和初级卵泡的卵母细胞中以及包括闭锁卵泡在内的卵泡各阶段的卵丘、颗粒细胞和卵泡膜细胞中观察到EGF-R免疫染色。在大型有腔卵泡中,卵泡膜细胞中的免疫染色强度比颗粒细胞中更明显,而卵母细胞的染色很少或未检测到。当用免疫前血清(EGF)替代一抗或用对照肽(EGF-R)预吸收时,未观察到免疫染色,证实了染色程序的特异性。(摘要截断于250字)

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