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Regional Differences in Penetration of the Protein Stabilizer Trimethoprim (TMP) in the Rat Central Nervous System
Frontiers in Molecular Neuroscience ( IF 3.5 ) Pub Date : 2020-08-11 , DOI: 10.3389/fnmol.2020.00167
Benjamin V. Ineichen , Serena Di Palma , Endre Laczko , Shane A. Liddelow , Susanne Neumann , Martin E. Schwab , Alice C. Mosberger

Regulating gene expression at the protein level is becoming increasingly important for answering basic questions in neurobiology. Several techniques using destabilizing domains (DD) on transgenes, which can be activated or deactivated by specific drugs, have been developed to achieve this goal. A DD from bacterial dihydrofolate reductase bound and stabilized by trimethoprim (TMP) represents such a tool. To control transgenic protein levels in the brain, the DD-regulating drugs need to have sufficient penetration into the central nervous system (CNS). Yet, very limited information is available on TMP pharmacokinetics in the CNS following systemic injection. Here, we performed a pharmacokinetic study on the penetration of TMP into different CNS compartments in the rat. We used mass spectrometry to measure TMP concentrations in serum, cerebrospinal fluid (CSF) and tissue samples of different CNS regions upon intraperitoneal TMP injection. We show that TMP quickly (within 10 min) penetrates from serum to CSF through the blood-CSF barrier. TMP also shows quick penetration into brain tissue but concentrations were an order of magnitude lower compared to serum or CSF. TMP concentration in spinal cord was lower than in any other analyzed CNS area. Nevertheless, effective levels of TMP to stabilize DDs can be reached in the CNS with half-lives around 2 h. These data show that TMP has good and fast penetration properties into the CNS and is therefore a valuable ligand for precisely controlling protein expression in the CNS in rodents.



中文翻译:

大鼠中枢神经系统中蛋白质稳定剂甲氧苄啶(TMP)渗透的区域差异。

在回答神经生物学的基本问题时,在蛋白质水平上调节基因表达变得越来越重要。已经开发了几种使用转基因上的去稳定结构域(DD)的技术来实现这一目标,这些技术可以通过特定的药物激活或去激活。由甲氧苄啶(TMP)结合并稳定化的细菌二氢叶酸还原酶的DD代表了这种工具。为了控制大脑中的转基因蛋白水平,调节DD的药物需要充分渗透到中枢神经系统(CNS)中。然而,全身注射后中枢神经系统中有关TMP药代动力学的信息非常有限。在这里,我们进行了关于TMP穿透大鼠不同CNS隔室的药代动力学研究。我们使用质谱法测量血清中的TMP浓度,腹膜内注射TMP后脑脊液(CSF)和不同中枢神经系统区域的组织样本。我们显示,TMP快速(在10分钟内)通过血液CSF屏障从血清渗透到CSF。TMP还显示快速渗透到脑组织中,但与血清或CSF相比,其浓度要低一个数量级。脊髓中的TMP浓度低于其他任何分析过的CNS区域。然而,在CNS中可以达到稳定DDs的有效TMP水平,其半衰期约为2小时。这些数据表明,TMP具有良好且快速的渗透特性,因此是精确控制啮齿动物中CNS中蛋白质表达的有价值的配体。我们显示,TMP快速(在10分钟内)通过血液CSF屏障从血清渗透到CSF。TMP还显示出迅速渗透到脑组织中,但与血清或CSF相比,浓度要低一个数量级。脊髓中的TMP浓度低于其他任何分析过的CNS区域。然而,在CNS中可以达到稳定DDs的有效TMP水平,其半衰期约为2小时。这些数据表明,TMP具有良好且快速的渗透特性,因此是精确控制啮齿动物中CNS中蛋白质表达的有价值的配体。我们显示,TMP快速(在10分钟内)通过血液CSF屏障从血清渗透到CSF。TMP还显示快速渗透到脑组织中,但与血清或CSF相比,其浓度要低一个数量级。脊髓中的TMP浓度低于其他任何分析过的CNS区域。然而,在CNS中可以达到稳定DDs的有效TMP水平,其半衰期约为2小时。这些数据表明,TMP具有良好且快速的渗透特性,因此是精确控制啮齿动物中CNS中蛋白质表达的有价值的配体。然而,在CNS中可以达到稳定DDs的有效TMP水平,其半衰期约为2小时。这些数据表明,TMP具有良好且快速的渗透特性,因此是精确控制啮齿动物中CNS中蛋白质表达的有价值的配体。然而,在CNS中可以达到稳定DDs的有效TMP水平,其半衰期约为2小时。这些数据表明,TMP具有良好和快速的渗透特性,因此是在啮齿动物中精确控制CNS中蛋白质表达的有价值的配体。

更新日期:2020-09-03
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