Division of Neurotoxicology, National Center for Toxicological Research, U.S. Food and Drug Administration, 3900 NCTR Rd, Jefferson, AR, 72079, USA.
Mol Neurobiol. 2018 Jan;55(1):122-129. doi: 10.1007/s12035-017-0724-y.
The 14-3-3 proteins are among the most abundant proteins expressed in the brain, comprising about 1% of the total amount of soluble brain proteins. Through phosphoserine- and phosphothreonine-binding motifs, 14-3-3 proteins regulate many signaling proteins and cellular processes including cell death. In the present study, we utilized a well-known kainic acid (KA)-induced excitotoxicity rat model and examined the expression of 14-3-3 and its isoforms in the frontal cortex of KA-treated and control animals. Among the different 14-3-3 isoforms, abundant levels of eta and tau were detected in the frontal cortex, followed by sigma, epsilon, and gamma, while the expression levels of alpha/beta and zeta/delta isoforms were low. Compared to the control animals, KA treatment induced a significant downregulation of the overall 14-3-3 protein level as well as the levels of the abundant isoforms eta, tau, epsilon, and gamma. We also investigated two 14-3-3-interacting proteins that are involved in the cell death process: Bcl-2-associated X (BAX) and extracellular signal-regulated kinase (ERK). Both BAX and phosphorylated ERK showed increased levels following KA treatment. Together, these findings demonstrate an abundance of several 14-3-3 isoforms in the frontal cortex and that KA treatment can cause a downregulation of 14-3-3 expression and an upregulation of 14-3-3-interacting proteins BAX and phospho-ERK. Thus, downregulation of 14-3-3 proteins could be one of the early molecular events associated with excitotoxicity. This could lead to subsequent upregulation of 14-3-3-binding proteins such as BAX and phospho-ERK that contribute to further downstream apoptosis processes, eventually leading to cell death. Maintaining sufficient levels of 14-3-3 expression and function may become a target of therapeutic intervention for excitotoxicity-induced neurodegeneration.
14-3-3 蛋白是大脑中表达最丰富的蛋白质之一,约占可溶性脑蛋白总量的 1%。通过磷酸丝氨酸和磷酸苏氨酸结合基序,14-3-3 蛋白调节许多信号蛋白和细胞过程,包括细胞死亡。在本研究中,我们利用了一种众所周知的红藻氨酸(KA)诱导的兴奋性毒性大鼠模型,检查了 KA 处理和对照动物额皮质中 14-3-3 及其同工型的表达。在不同的 14-3-3 同工型中,eta 和 tau 大量存在于额皮质中,其次是 sigma、epsilon 和 gamma,而 alpha/beta 和 zeta/delta 同工型的表达水平较低。与对照动物相比,KA 处理显著下调了总 14-3-3 蛋白水平以及丰富同工型 eta、tau、epsilon 和 gamma 的水平。我们还研究了两种参与细胞死亡过程的 14-3-3 相互作用蛋白:Bcl-2 相关 X(BAX)和细胞外信号调节激酶(ERK)。KA 处理后,BAX 和磷酸化 ERK 的水平均升高。总之,这些发现表明额皮质中有多种 14-3-3 同工型的丰富存在,KA 处理可导致 14-3-3 表达下调和 14-3-3 相互作用蛋白 BAX 和磷酸化 ERK 的上调。因此,14-3-3 蛋白的下调可能是与兴奋性毒性相关的早期分子事件之一。这可能导致随后上调 14-3-3 结合蛋白,如 BAX 和磷酸化 ERK,从而进一步促进下游细胞凋亡过程,最终导致细胞死亡。维持足够的 14-3-3 表达和功能水平可能成为治疗兴奋性毒性诱导的神经退行性变的靶点。