RNA-Seq analysis of knocking out the neuroprotective proton-sensitive GPR68 on basal and acute ischemia-induced transcriptome changes and signaling in mouse brain
| Autor: | Tao Wang, Guokun Zhou, Xiang-ming Zha |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Cell signaling Biochemistry Neuroprotection Article Brain Ischemia Receptors G-Protein-Coupled 03 medical and health sciences Endoplasmic reticulum chaperone complex 0302 clinical medicine Downregulation and upregulation Genetics Animals Receptor Molecular Biology Mice Knockout Neurons Chemistry Wild type Brain Long-term potentiation Cell biology Mice Inbred C57BL Disease Models Animal 030104 developmental biology Synaptic signaling Protons Transcriptome 030217 neurology & neurosurgery Biotechnology Signal Transduction |
| Zdroj: | FASEB J |
| ISSN: | 1530-6860 |
| Popis: | Brain acid signaling plays important roles in both physiological and disease conditions. One key neuronal metabotropic proton receptor in the brain is GPR68, which contributes to hippocampal long-term potentiation (LTP) and mediates neuroprotection in acidotic and ischemic conditions. Here, to gain greater understanding of GPR68 function in the brain, we performed mRNA-Seq analysis in mice. First, we studied sham-operated animals to determine baseline expression. Compared to wild type (WT), GPR68-/- (KO) brain downregulated genes that are enriched in Gene Ontology (GO) terms of misfolding protein binding, response to organic cyclic compounds, and endoplasmic reticulum chaperone complex. Next, we examined the expression profile following transient middle cerebral artery occlusion (tMCAO). tMCAO-upregulated genes cluster to cytokine/chemokine-related functions and immune responses, while tMCAO-downregulated genes cluster to channel activities and synaptic signaling. For proton-sensitive receptors, tMCAO downregulated ASIC1a and upregulated GPR4 and GPR65, but had no effect on ASIC2, PAC, or GPR68. GPR68 deletion did not alter the expression of these proton receptors, either at baseline or after ischemia. Lastly, we performed GeneVenn analysis of differential genes at baseline and post-tMCAO. Ischemia upregulated the expression of three hemoglobin genes, along with H2-Aa, Ppbp, Siglece, and Tagln, in WT but not in KO. Immunostaining showed that tMCAO-induced hemoglobin localized to neurons. Western blot analysis further showed that hemoglobin induction is GPR68-dependent. Together, these data suggest that GPR68 deletion at baseline disrupts chaperone functions and cellular signaling responses and imply a contribution of hemoglobin-mediated antioxidant mechanism to GPR68-dependent neuroprotection in ischemia. |
| Databáze: | OpenAIRE |
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