Stress-Induced Sex- and Cell Type-Specific Proteomic Changes in the Striatum: Impact on Substance Abuse
Cheng Jiang, Department of Psychiatry, Yale University
Substance use disorder (SUD) has been identified as a major national health emergency with a devastating impact on our society and public health. Substances of abuse, such as psychostimulants and opiates, exert their reinforcing effects by inducing neural alterations in the striatum and many of these changes occur in two main subtypes of medium spiny neurons (MSNs) which are differentiated by enrichment of dopamine receptor 1 (D1) or dopamine receptor 2 (D2) and by their respective projection pathways (striatonigral D1 direct pathway and striatopallidal D2 indirect pathway). Stress is a major risk factor for the development of SUD and addiction relapse, and significantly contributes to the sex differences in vulnerability to addiction. Stress-induced addiction is thought to be mediated by dysregulated synaptic plasticity in the striatum, resulting in altered synaptic signaling in a cell type-specific manner. Therefore, understanding how stress influences the proteome of the striatum in a sex- and cell type-specific manner is a crucial first step in identifying the protein networks that underlie synaptic alterations that may precipitate addictive behaviors, and could lead to the development of novel and effective treatment options for SUD.
With the state-of-the-art technologies and expertise provided by the Yale/NIDA Neuroproteomics Center, we propose to optimize a laser capture microdissection (LCM)-based proteomics workflow to identify sex- and cell type-specific proteomic changes in D1 and D2 neurons in the striatum of mice subjected to subchronic variable stress (SCVS). In Aim 1, we will refine a method using LCM to isolate D1 and D2 neurons from the striatum of Drd1-EGFP and Drd2-EGFP mice for proteomic analysis. In Aim 2, we will identify proteomic changes in the whole striatum tissue and in striatal D1 and D2 neurons following SCVS in both female and male mice. The proposed study will allow us to optimize a LCM-based proteomics workflow and generate original data characterizing the sex differences in stress-induced cell type-specific proteomic changes in the striatum. The novel protein targets identified in our discovery-based study may also encourage future hypothesis-driven investigations into sex- and cell type-specific signaling pathways that may precipitate increased vulnerability to substance abuse.
With the state-of-the-art technologies and expertise provided by the Yale/NIDA Neuroproteomics Center, we propose to optimize a laser capture microdissection (LCM)-based proteomics workflow to identify sex- and cell type-specific proteomic changes in D1 and D2 neurons in the striatum of mice subjected to subchronic variable stress (SCVS). In Aim 1, we will refine a method using LCM to isolate D1 and D2 neurons from the striatum of Drd1-EGFP and Drd2-EGFP mice for proteomic analysis. In Aim 2, we will identify proteomic changes in the whole striatum tissue and in striatal D1 and D2 neurons following SCVS in both female and male mice. The proposed study will allow us to optimize a LCM-based proteomics workflow and generate original data characterizing the sex differences in stress-induced cell type-specific proteomic changes in the striatum. The novel protein targets identified in our discovery-based study may also encourage future hypothesis-driven investigations into sex- and cell type-specific signaling pathways that may precipitate increased vulnerability to substance abuse.