Tissue-specific protein kinase C isoform expression in rat uterine tissue.
Tissue-specific protein kinase C isoform expression in rat uterine tissue.
复制标题
大鼠子宫组织中组织特异性蛋白激酶 C 异构体的表达。
DOI：
10.1016/s1071-5576(99)00035-0
复制
发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Phillippe,M
中科院分区：
文献类型：
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作者：
Kim,TT;Saunders,T;Bieber,E;Phillippe,M
目的：磷脂酰肌醇信号通路的激活在激动剂刺激的阶段性子宫肌收缩的产生中起关键作用。蛋白激酶C （PKC）是该信号通路的一个组成部分，先前已被证明对激动剂刺激的子宫肌收缩产生反馈抑制。本报告中描述的研究是为了调查大鼠子宫中几种PKC亚型的组织特异性表达。方法：取定时妊娠和正常骑行的成年雌性sd大鼠子宫组织。采用Vectastian ABC免疫染色技术和PKC异构体特异性多克隆抗体进行免疫组化研究。Western blot研究采用差速离心将肌组织分离成细胞质和膜组分。结果：这些研究证实了PKC-α、-β2、-δ、-η和-ζ在妊娠和发情大鼠肌层中有显著表达，而PKC-β1、-γ、-ε和-θ在妊娠和发情大鼠肌层中只有微量或无表达。PKC-α、-β2和-η亚型的表达在妊娠后期略有下降；相比之下，PKC-δ和-ζ在此期间保持稳定。免疫组化研究证实了近期妊娠大鼠子宫圆形和纵向平滑肌层中PKC-α、-β2、-δ、-η和-ζ亚型的表达。结论：综上所述，这些研究证实PKC的几种同工型在发情期和孕中期大鼠子宫组织中均有显著表达，其中以肌层平滑肌细胞表达最为突出。
Objective:Activation of the phosphatidylinositol signaling pathway plays a key role during the generation of agonist-stimulated phasic myometrial contractions. Protein kinase C (PKC), a component of this signaling pathway, has been previously shown to produce feedback inhibition of agonist-stimulated myometrial contractions. The studies described in this report were performed to survey the tissue-specific expression of several PKC isoforms in the rat uterus.Methods:Uterine tissue was obtained from timed pregnant and normally cycling adult female Sprague-Dawley rats. Immunohistochemical studies were performed using the Vectastian ABC immunostaining technique and PKC isoform-specific polyclonal antibodies. Western blot studies were peformed using myometrial tissue separated into cytosol and membrane fractions by differential centrifugation.Results:These studies confirmed significant expression of the PKC-α, -β2, -δ, -η, and -ζ isoforms in myometrium from pregnant and estrus rats, whereas only trace or no expression of the PKC-β1, -γ, -ε, and -θ isoforms was observed. Expression of the PKC-α, -β2, and -η isoforms decreased modestly during the latter days of gestation; in contrast, PKC-δ and -ζ remained stable during this period. The immunohistochemical studies confirmed expression of the PKC-α, -β2, -δ, -η, and -ζ isoforms in both circular and longitudinal smooth-muscle layers of the near-term pregnant rat uterus.Conclusion:In summary, these studies have confirmed significant levels of expression of several isoforms of PKC in estrus and near-term pregnant rat uterine tissue, which was most prominent in the smooth-muscle cells of the myometrium.