蛋白激酶 C 同工型 α、δ 和 ε 在不同发育阶段的小鼠卵巢中呈现差异性表达。
Protein kinase C isoforms α, δ and ε are differentially expressed in mouse ovaries at different stages of postnatal development.
出版信息
J Ovarian Res. 2014 Dec 10;7:117. doi: 10.1186/s13048-014-0117-z.
BACKGROUND
The protein kinase C (PKC) is a family of serine/threonine kinases that consists of 12 different isoforms. Since PKC isoform expressions are known to be specific for different cell types and postnatal developmental stages, we aimed to determine immunolocalizations and protein expression levels of different PKC isoforms in pre-pubertal, pubertal and adult mouse ovaries.
METHODS
Ovaries were obtained from postnatal day 1 (PND1) and PND7 of pre-pubertal, PND21 of pubertal and PND60 of adult mice. Immunolocalizations of PKCα, PKCδ and PKCε isoforms were determined and immunostainings in different cellular components of all follicular stages were evaluated by H-Score. PKCα, PKCδ and PKCε protein expression levels were determined by Western blot. The bands were quantified via ImageJ software. The data obtained from H-Score and ImageJ evaluations were analyzed by ANOVA statistical test.
RESULTS
PKCα immunostainings were more intense in oocytes when compared to granulosa and theca cells at different follicular stages of all groups. The Western blot analysis revealed that PKCα expression was significantly higher in PND60 adult ovaries. Conversely, PKCδ immunostainings were more intense in granulosa cells. According to the Western blot analysis, PKCδ protein expression was also higher in PND60 and significantly lower in PND1 ovaries. PKCε immunostaining was more apparent in oocytes. PKCε protein expression was significantly higher in adult PND60 and pubertal PND21 ovaries when compared to pre-pubertal PND7 and PND1 ovaries. Interestingly, PKCε immunostaining was significantly higher in primordial follicles, though PKCα and PKCδ immunostainings were more apparent in larger follicles. PKCα immunostainings of corpora lutea (CL) were significantly higher when compared to follicles in PND60 ovaries.
CONCLUSIONS
This study demonstrates that PKCα, PKCδ and PKCε isoforms are differentially expressed in particular cellular components of pre-pubertal, pubertal and adult mouse ovarian follicles. Therefore, we suggest that each PKC isoform has unique functions that are controlled by gonadotropin dependent mechanisms during follicular growth, oocyte maturation, ovulation and luteinization.
背景
蛋白激酶 C（PKC）是一个丝氨酸/苏氨酸激酶家族，由 12 种不同的同工型组成。由于 PKC 同工型的表达被认为是特定于不同的细胞类型和出生后发育阶段的，我们旨在确定不同 PKC 同工型在青春期前、青春期和成年小鼠卵巢中的免疫定位和蛋白表达水平。
方法
从青春期前的 PND1 和 PND7、青春期的 PND21 和成年的 PND60 小鼠中获得卵巢。通过 H 评分评估不同卵泡阶段所有细胞成分中的 PKCα、PKCδ 和 PKCε 同工型的免疫定位，并对免疫染色进行评估。通过 Western blot 确定 PKCα、PKCδ 和 PKCε 的蛋白表达水平。通过 ImageJ 软件对条带进行定量。通过方差分析统计检验分析来自 H 评分和 ImageJ 评估的数据。
结果
与不同卵泡阶段的颗粒细胞和膜细胞相比，PKCα 免疫染色在卵母细胞中更为强烈。Western blot 分析显示，PKCα 在 PND60 成年卵巢中的表达明显更高。相反，PKCδ 免疫染色在颗粒细胞中更为强烈。根据 Western blot 分析，PKCδ 蛋白表达在 PND60 和 PND1 卵巢中也较高，在 PND1 卵巢中显著降低。PKCε 免疫染色在卵母细胞中更为明显。与青春期前的 PND7 和 PND1 卵巢相比，PKCε 蛋白表达在成年的 PND60 和青春期的 PND21 卵巢中显著更高。有趣的是，尽管 PKCα 和 PKCδ 免疫染色在较大的卵泡中更为明显，但 PKCε 免疫染色在原始卵泡中明显更高。与 PND60 卵巢中的卵泡相比，黄体（CL）的 PKCα 免疫染色明显更高。
结论
本研究表明，PKCα、PKCδ 和 PKCε 同工型在青春期前、青春期和成年小鼠卵巢卵泡的特定细胞成分中差异表达。因此，我们认为每个 PKC 同工型都具有独特的功能，这些功能受促性腺激素依赖机制的控制，在卵泡生长、卵母细胞成熟、排卵和黄体化过程中发挥作用。
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