Broad spectrum proteomics analysis of the inferior colliculus following acute hydrogen sulfide exposure

Abstract

Acute exposure to high concentrations of HS causes severe brain injury and long-term neurological disorders, but the mechanisms involved are not known. To better understand the cellular and molecular mechanisms involved in acute HS-induced neurodegeneration we used a broad-spectrum proteomic analysis approach to identify key molecules and molecular pathways involved in the pathogenesis of acute HS-induced neurotoxicity and neurodegeneration. Mice were subjected to acute inhalation exposure of up to750 ppm of HS. HS induced behavioral deficits and severe lesions including hemorrhage in the inferior colliculus (IC). The IC was microdissected for proteomic analysis. Tandem mass tags (TMT) liquid chromatography mass spectrometry (LC-MS/MS)-based quantitative proteomics was applied for protein identification and quantitation. LC-MS/MS identified 598, 562, and 546 altered proteomic changes at 2 h, and on days 2 and 4 post-HS exposure, respectively. Of these, 77 proteomic changes were statistically significant at any of the 3 time points. Mass spectrometry data were subjected to Perseus 1.5.5.3 statistical analysis, and gene ontology heat map clustering. Expressions of several key molecules were verified to confirm HS-dependent proteomics changes. Webgestalt pathway overrepresentation enrichment analysis with Panther engine revealed HS exposure disrupted several biological processes including metabotropic glutamate receptor group 1 and inflammation mediated by chemokine and cytokine signaling pathways among others. Further analysis showed that energy metabolism, integrity of blood-brain barrier, hypoxic, and oxidative stress signaling pathways were also implicated. Collectively, this broad-spectrum proteomics data has provided important clues to follow up in future studies to further elucidate mechanisms of HS-induced neurotoxicity.

Transcriptomic profile analysis of brain inferior colliculus following acute hydrogen sulfide exposure

“…F3, plat, and plaur were upregulated 2 h after a single acute H 2 S exposure and after two acute exposures (early response), while alpha-2-macroglobulin (A2m) and von Willebrand factor (Vwf) were upregulated from early response (2 exposures to H 2 S) to late response (4 exposures to H 2 S). These results further support our previous findings that H 2 S exposure may induce hemorrhage in the IC, suggesting a breach of the blood brain barrier may play a role in H 2 S-induced neurotoxicity (Kim et al 2018a).…”

“…The late response was measured following 4 days of acute H 2 S exposure. Mice exposed to H 2 S showed motorbehavioral deficits, respiratory depression, seizures and knock-down phenotype as previously reported ((Anantharam et al 2017;Kim et al 2018b)). C57 black mice were exposed via whole body exposure to 700 ppm H 2 S. A scheme of exposure paradigm was shown (A).…”

“…In our previous studies we have demonstrated that H 2 S-inhibits cytochrome c oxidase (Anantharam et al 2017); moreover, H 2 S exposure also led to the generation of hydroxyl radicals (Jiang et al 2016) all of which contribute to oxidative stress. In our previous work, we have reported that H 2 S exposure activated the Nrf2 signaling pathway (Kim et al 2018a), further supporting the hypothesis of H 2 S-induced oxidative stress as a toxic mechanism. MAF BZIP transcription factor F (Maff) is a basic leucine zipper transcription factor and forms a heterodimer with Nrf2 to induce antioxidant or electrophile response elements (AREs) (Katsuoka et al 2005).…”

“…We have previously shown that H 2 S exposure activated the hypoxic signaling pathway and upregulated Hif-1α (Kim et al 2018a). The Akt signaling pathway has previously been shown to be activated under hypoxic conditions and reportedly plays an important role in survival of neuronal cells (Alvarez-Tejado et al 2001); it also mediates the Hif-1 signaling pathway (Kilic-Eren et al 2013).…”

H2S Exposure Induces Autophagy Injury in The Jejunum of Broilers Through miR-181a-PKC-δ Axis Mediated ER Stress

“…Kim et al identi ed the acute neurodegeneration of cells induced by H 2 S and the molecular mechanisms by analyzing the key molecules and the molecular pathways in the broad-spectrum proteome (Kim et al, 2018). Similarly, it has been reported that DEPs associated with autophagy death pathway by using proteomic analysis increased signi cantly in AS-6 treated cells, therefore, which was induced by ER stress (Kang et al, 2012).…”