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钙敏感受体蛋白在人卵母细胞中的定位及定量表达

Localization and quantitative expression of the calcium-sensing receptor protein in human oocytes.

作者信息

Dell'Aquila Maria Elena, De Santis Teresa, Cho Yoon Sung, Reshkin Stephan Joel, Caroli Anna Maria, Maritato Filippo, Minoia Paolo, Casavola Valeria

机构信息

Department of Animal Production, University of Bari, Bari, Italy.

出版信息

Fertil Steril. 2006 Apr;85 Suppl 1:1240-7. doi: 10.1016/j.fertnstert.2005.11.033.

Abstract

OBJECTIVE

To investigate the expression of the calcium-sensing receptor (CaSR) protein in human oocytes at the germinal vesicle (GV) and metaphase I (MI) and II (MII) stages.

DESIGN

Prospective study.

SETTING

Academic basic research laboratory and hospital-based fertility center.

PATIENT(S): Immature and supernumerary mature oocytes (n = 118) excluded from intracytoplasmic sperm injection treatment.

INTERVENTION(S): Immunofluorescence and Western blot with a primary antibody against human CaSR. Confocal laser scanning microscopy (CLSM) together with quantitative image analysis used to measure the fluorescence intensity variations in oocytes at GV, MI, and MII stages.

MAIN OUTCOME MEASURE(S): The CaSR expression pattern as evaluated by immunostaining in denuded oocytes and cumulus cells, CLSM, and three-dimensional image reconstructions; quantitative analysis at the equatorial plane of the oocyte.

RESULT(S): We identified CaSR in human oocytes and cumulus cells. The fluorescence intensity within the oocyte varied with the developmental stage, with the greatest increase at the MI stage.

CONCLUSION(S): The present study demonstrates for the first time the expression and localization of CaSR protein in human oocytes. Increased CaSR protein expression in the MI stage suggests that it may be involved in the regulation of human oocyte development and maturation.

摘要

目的

研究钙敏感受体(CaSR)蛋白在人类生发泡(GV)期、减数分裂中期I(MI)和减数分裂中期II(MII)卵母细胞中的表达情况。

设计

前瞻性研究。

地点

学术基础研究实验室和医院生殖中心。

患者

因胞浆内单精子注射治疗而被排除的未成熟及多余的成熟卵母细胞(n = 118)。

干预措施

使用抗人CaSR的一抗进行免疫荧光和蛋白质免疫印迹。共聚焦激光扫描显微镜(CLSM)结合定量图像分析,用于测量GV期、MI期和MII期卵母细胞中的荧光强度变化。

主要观察指标

通过对裸卵和卵丘细胞进行免疫染色、CLSM及三维图像重建来评估CaSR的表达模式;在卵母细胞赤道面进行定量分析。

结果

我们在人类卵母细胞和卵丘细胞中鉴定出了CaSR。卵母细胞内的荧光强度随发育阶段而变化,在MI期增加最为显著。

结论

本研究首次证明了CaSR蛋白在人类卵母细胞中的表达及定位。MI期CaSR蛋白表达增加表明其可能参与人类卵母细胞发育和成熟的调控。

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