Identification and distribution of a novel platelet-derived growth factor receptor beta variant: effect of retinoic acid and involvement in cell differentiation.
Identification and distribution of a novel platelet-derived growth factor receptor beta variant: effect of retinoic acid and involvement in cell differentiation.
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DOI：
10.1210/en.2006-1206
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发表时间：
2007-05
期刊：
影响因子：
4.8
通讯作者：
Yan Wang;M. Culty
中科院分区：
文献类型：
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作者：
Yan Wang;M. Culty
我们以前已经表明，新生儿睾丸生殖细胞表达血小板衍生生长因子受体（PDGFR）α和β。我们报告了一种新的PDGFR β（V1-PDGFR β）转录本在3-d龄大鼠睾丸的生殖细胞中的表达。V1-PDGFR β核苷酸序列跨越PDGFR β基因的内含子6至外显子23，并且预测编码缺乏部分细胞外结构域的蛋白质。V1-PDGFR β转录物优先在发育中的性腺中表达。胚胎畸胎癌F9细胞，其中分化是由视黄酸（RA）驱动，表达V1-PDGFR β，但不表达野生型PDGFR β。绿色荧光蛋白标记的V1-PDGFR β主要定位于F9、MA-10和COS-1细胞的胞质溶胶中。FLAG和绿色荧光蛋白标记的V1-PDGFR β显示酪氨酸激酶活性，并含有磷酸酪氨酸残基，表明V1-PDGFR β是一种胞质酪氨酸激酶。用RA处理F9细胞诱导V1-PDGFR β基因表达，伴随着形态学变化和IV型胶原和层粘连蛋白B1的mRNA表达增加，表明V1-PFGR β参与细胞分化。同样，用RA处理出生后第3天大鼠生殖细胞诱导V1-PDGFR β表达呈剂量依赖性增加，同时c-kit和Stra 8（更分化生殖细胞的标志物）增加，GFR α 1（精原干细胞的标志物）随之减少。然而，过量的V1-PDGFR β抑制RA介导的IV型胶原和层粘连蛋白B1的表达，并改变F9细胞中RA依赖性和RA非依赖性的形态学变化，同时增加细胞存活。这些结果表明，V1-PDGFR β的表达在分化过程中受到严格调节，并且它可能在生殖细胞分化中起积极作用。
We have shown previously that neonatal testicular gonocytes express platelet-derived growth factor receptors (PDGFR) alpha and beta. We report the expression of a novel PDGFRbeta (V1-PDGFRbeta) transcript in gonocytes of 3-d-old rat testes. V1-PDGFRbeta nucleotide sequence spans from intron 6 to exon 23 of the PDGFRbeta gene, and is predicted to encode a protein lacking part of the extracellular domain. V1-PDGFRbeta transcripts are expressed preferentially in developing gonads. The embryonic teratocarcinoma F9 cells, in which differentiation is driven by retinoic acid (RA), express V1-PDGFRbeta, but not wild-type PDGFRbeta. Green fluorescent protein-tagged V1-PDGFRbeta localized mainly in cytosol of F9, MA-10, and COS-1 cells. FLAG and green fluorescent protein-tagged V1-PDGFRbeta displayed tyrosine kinase activities and contain phosphotyrosine residues, suggesting that V1-PDGFRbeta is a cytosolic tyrosine kinase. Treatment of F9 cells with RA induced V1-PDGFRbeta gene expression, concomitant with changes in morphology and increased mRNA expression of collagen IV and laminin B1, suggesting that V1-PFGRbeta is involved in cell differentiation. Similarly, treatment of postnatal d 3 rat gonocytes with RA induced a dose-dependent increase in V1-PDGFRbeta expression together with an increase in c-kit and Stra8, markers of more differentiated germ cells and a concomitant decrease in GFRalpha1, a marker of spermatogonial stem cells. However, an excess of V1-PDGFRbeta inhibited RA-mediated collagen IV and laminin B1 expression and altered both RA-dependent and RA-independent morphological changes in F9 cells, while increasing cell survival. These results suggest that the expression of V1-PDGFRbeta is tightly regulated during differentiation and that it may play an active role in germ cell differentiation.