Bioorthogonal chemical imaging of metabolic activities in live mammalian hippocampal tissues with stimulated Raman scattering
| Autor: | Wei Min, Michael R. Lamprecht, Fanghao Hu, Lu Wei, Barclay Morrison |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Brain--Imaging Protein metabolism Hippocampal formation Spectrum Analysis Raman Hippocampus Article Mass Spectrometry Mass spectrometry imaging Choline Activation Metabolic 03 medical and health sciences chemistry.chemical_compound Nucleic Acids Brain Injuries Traumatic Animals Phospholipids Cell metabolism Microscopy Multidisciplinary Dentate gyrus Fatty Acids Neurogenesis Brain Lipid metabolism Deuterium Lipid Metabolism Rats Chemistry 030104 developmental biology chemistry Alkynes Raman spectroscopy Nucleic acid Biophysics Bioorthogonal chemistry Biomedical engineering |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/srep39660 |
| Popis: | Brain is an immensely complex system displaying dynamic and heterogeneous metabolic activities. Visualizing cellular metabolism of nucleic acids, proteins, and lipids in brain with chemical specificity has been a long-standing challenge. Recent development in metabolic labeling of small biomolecules allows the study of these metabolisms at the global level. However, these techniques generally require nonphysiological sample preparation for either destructive mass spectrometry imaging or secondary labeling with relatively bulky fluorescent labels. In this study, we have demonstrated bioorthogonal chemical imaging of DNA, RNA, protein and lipid metabolism in live rat brain hippocampal tissues by coupling stimulated Raman scattering microscopy with integrated deuterium and alkyne labeling. Heterogeneous metabolic incorporations for different molecular species and neurogenesis with newly-incorporated DNA were observed in the dentate gyrus of hippocampus at the single cell level. We further applied this platform to study metabolic responses to traumatic brain injury in hippocampal slice cultures, and observed marked upregulation of protein and lipid metabolism particularly in the hilus region of the hippocampus within days of mechanical injury. Thus, our method paves the way for the study of complex metabolic profiles in live brain tissue under both physiological and pathological conditions with single-cell resolution and minimal perturbation. |
| Databáze: | OpenAIRE |
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