Identification of phospholipase C β isoforms and their location in cultured vascular smooth muscle cells of pig, human and rat
Identification of phospholipase C β isoforms and their location in cultured vascular smooth muscle cells of pig, human and rat
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DOI：
10.1016/s0008-6363(98)00193-x
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发表时间：
1998-12-01
影响因子：
10.8
通讯作者：
Rix, C
中科院分区：
文献类型：
--
作者：
Blayney, L;Gapper, P;Rix, C
目的：磷脂酶C（PLC）β在猪主动脉血管平滑肌中有4种亚型。目的是确定所有四种PLC β亚型是否通常在三种物种（即猪、人和大鼠）的血管平滑肌细胞（VSMC）中表达，以及各个亚型是否具有独特的细胞内分布。方法：血管平滑肌细胞培养来自猪和大鼠主动脉和人肾动脉细胞系的外植体。使用蛋白质印迹法解析PLC β同种型含量。免疫细胞化学和共聚焦显微镜观察细胞内定位。结果：所有三种物种均表达PLC β（1）、β（2）、β（3）和β（4）。在所有物种中，PLC β（1）在整个细胞质中显示出集中的病灶; PLC β（2）显示出主要在细胞周边或高尔基体中的点状模式，这取决于所使用的抗体; PLC β（3）也是胞质的，但显示出与PLC β（1）不同的模式，PLC β（4）是胞质的，除了在猪静止细胞中，在那里它与细胞膜交叉处的丝状结构相关。结论：三种不同物种的VSMC表达所有四种PLC β亚型。每种亚型都有一个独特的和一致的分布特征，这是所有物种普遍存在的。(C)1998年爱思唯尔科学BN。All rights reserved.
Objectives: Four phospholipase C (PLC) beta isoforms have been described in pig aortic vascular smooth muscle. The aim was to determine if all four PLC beta isoforms are commonly expressed in vascular smooth muscle cells (VSMC) of three species, i.e. pig, human and rat, and if the individual isoforms had distinctive intracellular distributions. Methods: Vascular smooth muscle cell cultures were derived from explants of porcine and rat aorta and a human renal artery cell line. PLC beta isoform content was resolved using Western blotting. Intracellular location was determined by immunocytochemistry and confocal microscopy. Results: All three species expressed PLCs beta(1), beta(2), beta(3) and beta(4). In all species, PLC beta(1) demonstrated foci of concentration throughout the cytoplasm; PLC beta(2) demonstrated a punctate pattern that was principally at the cell periphery or was in the Golgi, depending upon the antibody used; PLC beta(3) was, also cytoplasmic but showed a different pattern from PLC beta(1) and PLC beta(4) was cytoplasmic, except in pig quiescent cells, where it was associated with filamentous structures at the intersection with the plasma membrane. Conclusions: VSMCs of three different species express all four PLC beta isoforms. Each isoform has a unique and consistent signature of distribution that is generally common to all species. (C) 1998 Elsevier Science BN. All rights reserved.