Dopamine neuron-specific LRRK2 G2019S effects on gene expression revealed by translatome profiling
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Published on: December 14, 2017
Judit Pallos1, Sophia Jeng2, Shannon McWeeney3
1Jungers Center for Neurosciences, Department of Neurology, Oregon Health and Science University, Portland, OR, USA.
Abstract:
Leucine-rich repeat kinase 2 (LRRK2) mutations are the most common genetic cause of late-onset Parkinson's disease. The pathogenic G2019S mutation enhances LRRK2 kinase activity and induces neurodegeneration in C. elegans, Drosophila and rodent models through unclear mechanisms. Gene expression profiling has the potential to provide detailed insight into the biological pathways modulated by LRRK2 kinase activity. Prior in vivo studies have surveyed the effects of LRRK2 G2019S on genome-wide mRNA expression in complex brain tissues with high cellular heterogeneity, limiting their power to detect more restricted gene expression changes occurring in a cell type-specific manner. Here, we used translating ribosome affinity purification (TRAP) coupled to RNA-seq to profile dopamine neuron-specific gene expression changes caused by LRRK2 G2019S in the Drosophila CNS. A number of genes were differentially expressed in the presence of mutant LRRK2 that represent a broad range of molecular functions including DNA repair (RfC3), mRNA metabolism and translation (Ddx1 and lin-28), calcium homeostasis (MCU), and other categories (Ugt37c1, disp, l(1)G0196, CG6602, CG1126 and CG11068). Further analysis on a subset of these genes revealed that LRRK2 G2019S did not alter their expression across the whole brain, consistent with dopamine neuron-specific effects uncovered by the TRAP approach that may yield insight into the neurodegenerative process. To our knowledge, this is the first study to profile the effects of LRRK2 G2019S specifically on DA neuron gene expression in vivo. Beyond providing a set of differentially expressed gene candidates relevant to LRRK2, we demonstrate the effective use of TRAP to perform high-resolution assessment of dopamine neuron gene expression for the study of PD.
Insights
Leucine-rich repeat kinase 2 (LRRK2) mutations, a common cause of Parkinson's disease, were studied for their effects on dopamine neuron gene expression. This research utilized translating ribosome affinity purification (TRAP) to identify specific gene changes in vivo.
Area of Science:
- Neuroscience
- Genetics
- Molecular Biology
Background:
- Leucine-rich repeat kinase 2 (LRRK2) mutations are the primary genetic cause of late-onset Parkinson's disease (PD).
- The pathogenic G2019S LRRK2 mutation increases kinase activity, leading to neurodegeneration via poorly understood mechanisms.
- Previous studies examining LRRK2 G2019S effects on gene expression were limited by the cellular heterogeneity of brain tissues.
Purpose of the Study:
- To investigate dopamine neuron-specific gene expression changes induced by the LRRK2 G2019S mutation in vivo.
- To identify novel gene candidates and pathways affected by LRRK2 kinase activity in dopamine neurons.
- To demonstrate the utility of translating ribosome affinity purification (TRAP) for cell-type-specific gene expression analysis in Parkinson's disease research.
Main Methods:
- Utilized translating ribosome affinity purification (TRAP) coupled with RNA sequencing (RNA-seq) in a Drosophila model.
- Focused gene expression profiling specifically within dopamine neurons of the central nervous system (CNS).
- Analyzed differentially expressed genes to understand molecular functions and pathways impacted by mutant LRRK2.
Main Results:
- Identified a set of differentially expressed genes in dopamine neurons due to LRRK2 G2019S, including those involved in DNA repair, mRNA metabolism, translation, and calcium homeostasis.
- Confirmed that LRRK2 G2019S-induced expression changes were specific to dopamine neurons and not observed in whole-brain analyses.
- This study represents the first in vivo profiling of LRRK2 G2019S effects on dopamine neuron gene expression.
Conclusions:
- The TRAP approach provides high-resolution, cell-type-specific insights into gene expression changes in Parkinson's disease models.
- Differentially expressed genes identified offer potential targets for understanding LRRK2-mediated neurodegeneration.
- This work highlights the importance of cell-type-specific analysis for dissecting complex neurological disorders like Parkinson's disease.
