转化分析揭示了左旋多巴诱导的异动症小鼠模型中直接和间接通路中基因表达的新变化。
Translational profiling reveals novel gene expression changes in the direct and indirect pathways in a mouse model of levodopa induced dyskinesia.
作者信息
Jafri Sabika, Ghani Mahdi, Stickle Natalie, Virtanen Carl, Hazrati Lili-Naz, Visanji Naomi P
机构信息
Krembil Brain Institute, University Health Network, Toronto, ON, Canada.
Tanz Centre for Research in Neurodegenerative Disease, University of Toronto, Krembil Discovery Tower, Toronto, ON, Canada.
出版信息
Front Cell Neurosci. 2025 Mar 12;18:1477511. doi: 10.3389/fncel.2024.1477511. eCollection 2024.
INTRODUCTION
The molecular mechanisms underlying L-dihydroxyphenylalanine (LDOPA) induced dyskinesia in Parkinson's disease are poorly understood. Here we employ two transgenic mouse lines, combining translating ribosomal affinity purification (TRAP) with bacterial artificial chromosome expression (Bac), to selectively isolate RNA from either DRD1A expressing striatonigral, or DRD2 expressing striatopallidal medium spiny neurons (MSNs) of the direct and indirect pathways respectively, to study changes in translational gene expression following repeated LDOPA treatment.
METHODS
6-OHDA lesioned DRD1A and DRD2 BacTRAP mice were treated with either saline or LDOPA bi-daily for 21 days over which time they developed abnormal involuntary movements reminiscent of dyskinesia. On day 22, all animals received LDOPA 40min prior to sacrifice. The striatum of the lesioned hemisphere was dissected and subject to TRAP. Extracted ribosomal RNA was amplified, purified, and gene expression was quantified using microarray.
RESULTS
One hundred ninety-five significantly varying transcripts were identified among the four treatment groups. Pathway analysis revealed an overrepresentation of calcium signaling and long-term potentiation in the DRD1A expressing MSNs of the direct pathway, with significant involvement of long-term depression in the DRD2 expressing MSNs of the indirect pathway following chronic treatment with LDOPA. Several MAPK associated genes (, and ) differentiated the direct and indirect pathways following both acute and chronic LDOPA treatment. However, the MAPK pathway activator was downregulated in the indirect pathway and upregulated in the direct pathway, strongly suggesting a role for in regulating the opposing effects of LDOPA on these two pathways in dyskinesia.
DISCUSSION
Future studies will assess the potential of targeting these genes and pathways to prevent the development of LDOPA-induced dyskinesia.
引言
左旋二羟基苯丙氨酸（L-DOPA）诱发帕金森病异动症的分子机制目前仍知之甚少。在此，我们使用两种转基因小鼠品系，将翻译核糖体亲和纯化（TRAP）与细菌人工染色体表达（Bac）相结合，分别从直接通路中表达DRD1A的纹状体黑质中型多棘神经元（MSNs）或间接通路中表达DRD2的纹状体苍白球MSNs中选择性分离RNA，以研究重复给予L-DOPA治疗后翻译基因表达的变化。
方法
用6-羟基多巴胺（6-OHDA）损伤DRD1A和DRD2 BacTRAP小鼠，每天两次分别给予生理盐水或L-DOPA，持续21天，在此期间它们出现了类似于异动症的异常不自主运动。在第22天，所有动物在处死前40分钟接受L-DOPA。解剖损伤半球的纹状体并进行TRAP。提取的核糖体RNA经扩增、纯化，然后使用微阵列对基因表达进行定量分析。
结果
在四个治疗组中鉴定出195个有显著差异的转录本。通路分析显示，在直接通路中表达DRD1A的MSNs中，钙信号传导和长时程增强作用显著富集，而在间接通路中表达DRD2的MSNs中，长期给予L-DOPA治疗后长时程抑制作用显著参与其中。在急性和慢性L-DOPA治疗后，几个与丝裂原活化蛋白激酶（MAPK）相关的基因（ 、 和 ）区分了直接通路和间接通路。然而，MAPK通路激活剂 在间接通路中下调，在直接通路中上调，这强烈表明 在调节L-DOPA对异动症中这两条通路的相反作用方面发挥作用。
讨论
未来的研究将评估靶向这些基因和通路以预防L-DOPA诱发异动症发生的潜力。
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