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胎盘57千道尔顿钙结合蛋白:滋养层细胞钙转运中表达与功能的调节

Placental 57-kDa Ca(2+)-binding protein: regulation of expression and function in trophoblast calcium transport.

作者信息

Hershberger M E, Tuan R S

机构信息

Department of Orthopaedic Surgery, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.

出版信息

Dev Biol. 1998 Jul 1;199(1):80-92. doi: 10.1006/dbio.1998.8926.

Abstract

During gestation, transport by placental trophoblasts is solely responsible for nutrient supply to the developing fetus. The calcium (Ca) transport machinery of the placenta thus represents the primary tissue site for regulating fetal Ca homeostasis. The exact mechanism of trophoblast Ca transport is not known. However, there is evidence suggesting that a developmentally expressed cytosolic, trophoblast-specific, high M(r) 57-kDa Ca-binding protein (CaBP) plays an important role in regulating and/or shuttling cytosolic Ca. We report here the cloning of a full-length cDNA of the mouse CaBP which shows significant homology with calreticulin, an endoplasmic reticulum-associated Ca binding protein. The functional role of CaBP in cellular Ca handling was investigated using a trophoblastic cell line, Rcho-1, derived from a rat choriocarcinoma. Upon differentiation, Rcho-1 cells exhibit enhanced Ca uptake compared to undifferentiated Rcho-1 stem cells, and CaBP expression is upregulated. To analyze the regulation of CaBP expression, placenta organ cultures and Rcho-1 cells were treated for 48 h in vitro with a series of agents implicated in Ca homeostasis. In both placenta organ cultures and undifferentiated as well as differentiated Rcho-1 cells, treatment with 1,25-dihydroxy vitamin D3, estrogen, parathyroid hormone (PTH), parathyroid hormone-related protein (PTHrP 1-34), and Ca had no effect on CaBP mRNA and protein levels, which were significantly stimulated by PTHrP 67-84. PTHrP 67-84-treated Rcho-1 cells also exhibited higher Ca uptake activity than untreated control cells. The upregulation of CaBP expression during and/or following the differentiation of Rcho-1 cells into trophoblastic giant cells supports the importance of CaBP in trophoblast maturation and the validity of the Rcho-1 rat model cell system. In addition, the action of PTHrP on placental trophoblast Ca transport is likely to involve the regulation of CaBP expression to handle the increasing Ca requirements of the developing fetus.

摘要

在妊娠期间,胎盘滋养层细胞的转运是发育中胎儿营养供应的唯一负责途径。因此,胎盘的钙(Ca)转运机制是调节胎儿钙稳态的主要组织部位。滋养层细胞钙转运的确切机制尚不清楚。然而,有证据表明,一种在发育过程中表达的胞质、滋养层特异性、高Mr 57-kDa钙结合蛋白(CaBP)在调节和/或穿梭胞质钙方面发挥重要作用。我们在此报告了小鼠CaBP全长cDNA的克隆,该克隆与钙网蛋白(一种内质网相关的钙结合蛋白)具有显著同源性。使用源自大鼠绒毛膜癌的滋养层细胞系Rcho-1研究了CaBP在细胞钙处理中的功能作用。与未分化的Rcho-1干细胞相比,Rcho-1细胞在分化时表现出增强的钙摄取,并且CaBP表达上调。为了分析CaBP表达的调节,胎盘器官培养物和Rcho-1细胞在体外用一系列与钙稳态有关的试剂处理48小时。在胎盘器官培养物以及未分化和分化的Rcho-1细胞中,用1,25-二羟基维生素D3、雌激素、甲状旁腺激素(PTH)、甲状旁腺激素相关蛋白(PTHrP 1-34)和钙处理对CaBP mRNA和蛋白水平没有影响,而PTHrP 67-84则显著刺激了它们。用PTHrP 67-84处理的Rcho-1细胞也表现出比未处理的对照细胞更高的钙摄取活性。Rcho-1细胞分化为滋养层巨细胞期间和/或之后CaBP表达的上调支持了CaBP在滋养层成熟中的重要性以及Rcho-1大鼠模型细胞系统的有效性。此外,PTHrP对胎盘滋养层细胞钙转运的作用可能涉及CaBP表达的调节,以满足发育中胎儿不断增加的钙需求。

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