Prenatal methadone exposure selectively alters protein expression in primary motor cortex: Implications for synaptic function

Abstract

As problematic opioid use has reached epidemic levels over the past 2 decades, the annual prevalence of opioid use disorder (OUD) in pregnant women has also increased 333%. Yet, how opioids affect the developing brain of offspring from mothers experiencing OUD remains understudied and not fully understood. Animal models of prenatal opioid exposure have discovered many deficits in the offspring of prenatal opioid exposed mothers, such as delays in the development of sensorimotor function and long-term locomotive hyperactivity. In attempt to further understand these deficits and link them with protein changes driven by prenatal opioid exposure, we used a mouse model of prenatal methadone exposure (PME) and preformed an unbiased multi-omic analysis across many sensoriomotor brain regions known to interact with opioid exposure. The effects of PME exposure on the primary motor cortex (M1), primary somatosensory cortex (S1), the dorsomedial striatum (DMS), and dorsolateral striatum (DLS) were assessed using quantitative proteomics and phosphoproteomics. PME drove many changes in protein and phosphopeptide abundance across all brain regions sampled. Gene and gene ontology enrichments were used to assess how protein and phosphopeptide changes in each brain region were altered. Our findings showed that M1 was uniquely affected by PME in comparison to other brain regions. PME uniquely drove changes in M1 glutamatergic synapses and synaptic function. Immunohistochemical analysis also identified anatomical differences in M1 for upregulating the density of glutamatergic and downregulating the density of GABAergic synapses due to PME. Lastly, comparisons between M1 and non-M1 multi-omics revealed conserved brain wide changes in phosphopeptides associated with synaptic activity and assembly, but only specific protein changes in synapse activity and assembly were represented in M1. Together, our studies show that lasting changes in synaptic function driven by PME are largely represented by protein and anatomical changes in M1, which may serve as a starting point for future experimental and translational interventions that aim to reverse the adverse effects of PME on offspring.

The impact of abstinence from chronic alcohol consumption on the mouse striatal proteome: sex and subregion-specific differences

“…All proteomics experiments were performed in collaboration with the Indiana University Proteomics Core similarly to previous work done in our lab ( Grecco et al, 2021 ; Grecco et al, 2022 ; Haggerty et al, 2023 ).…”

“…We prepared proteins in the same way as our previous works ( Grecco et al, 2021 ; Grecco et al, 2022 ; Haggerty et al, 2023 ): Flash frozen brain sections were homogenized in 150–170 µL of 8 M urea in 100 mM Tris, pH 8.5 using a Bioruptor ® sonication system (Diagenode Inc. United States, North America Cat No: B01020001) with 30 s/30 s on/off cycles for 20 min in a water bath at 4°C. After subsequent centrifugation at 10,000 rcf for 20 min, protein concentrations were determined by Bradford protein assay (BioRad Cat No: 5000006).…”

“…Since data dependent mass spectrometry can be stochastic in which specific peptides are selected for isolation, fragmentation and thus quantification, we created a trigger channel ( Peck Justice et al, 2021 ) containing peptides from a selection of inflammatory proteins of interest identified by Western blot in our prior work ( Blaker and Yamamoto, 2018 ). We selected peptides from previous mass spectrometry analysis ( Grecco et al, 2021 ; Grecco et al, 2022 ; Haggerty et al, 2023 ), and when not previously identified by mass spectrometry, from high scoring peptides on peptideatlas.org ( Desiere et al, 2006 ). Peptides were purchased from Biosynth, and a 100 µg of an equimolar solution of 8 peptides were labeled with TMTPro channel 135N.…”

Traditional Chinese medicines and natural products targeting immune cells in the treatment of metabolic-related fatty liver disease

“…On the one hand, they can act as anti-inflammatory agents by binding to specific receptors, such as FXR and TGR5, on immune cells and hepatocytes (Chiang and Ferrell, 2020). Activation of these receptors inhibits the production of pro-inflammatory cytokines and promotes the expression of antiinflammatory genes (Haggerty et al, 2023). On the other hand, bile acids can also promote inflammation by disrupting the gut microbiota and increasing the permeability of the intestinal Frontiers in Pharmacology frontiersin.org barrier (Haggerty et al, 2023).…”

“…Activation of these receptors inhibits the production of pro-inflammatory cytokines and promotes the expression of antiinflammatory genes (Haggerty et al, 2023). On the other hand, bile acids can also promote inflammation by disrupting the gut microbiota and increasing the permeability of the intestinal Frontiers in Pharmacology frontiersin.org barrier (Haggerty et al, 2023). This leads to the translocation of bacterial products, such as lipopolysaccharides (LPS), into the bloodstream, which triggers an immune response and promotes liver inflammation (Carpino et al, 2020).…”