Spatial sterol metabolism unveiled by stimulated Raman imaging

Yongqing Zhang; Yihui Zhou; Wen Fang; Hanlin Zhu; Cunqi Ye; Delong Zhang; Hyeon Jeong Lee

doi:10.3389/fchem.2023.1166313

Spatial sterol metabolism unveiled by stimulated Raman imaging

Front Chem. 2023 Mar 29:11:1166313. doi: 10.3389/fchem.2023.1166313. eCollection 2023.

Authors

Yongqing Zhang¹, Yihui Zhou², Wen Fang³, Hanlin Zhu¹, Cunqi Ye³, Delong Zhang¹, Hyeon Jeong Lee²

Affiliations

¹ Zhejiang Province Key Laboratory of Quantum Technology and Device, Department of Physics, Interdisciplinary Centre for Quantum Information, Zhejiang University, Hangzhou, China.
² Key Laboratory for Biomedical Engineering of Ministry of Education, College of Biomedical Engineering and Instrument Science, Zhejiang University, Hangzhou, China.
³ Zhejiang Provincial Key Laboratory for Cancer Molecular Cell Biology, Life Sciences Institute, Zhejiang University, Hangzhou, China.

Abstract

Spatiotemporal dynamics of small-molecule metabolites have gained increasing attention for their essential roles in deciphering the fundamental machinery of life. However, subcellular-level regulatory mechanisms remain less studied, particularly due to a lack of tools to track small-molecule metabolites. To address this challenge, we developed high-resolution stimulated Raman scattering (SRS) imaging of a genetically engineered model (GEM) to map metabolites in subcellular resolution. As a result, an unexpected regulatory mechanism of a critical metabolite, sterol, was discovered in yeast by amplifying the strength of vibrational imaging by genetic modulation. Specifically, isozymes of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) were evident to promote ergosterol distribution to distinct subcellular locations, where ergosterol was enriched by a local HMGR-directed synthesis. The heterogeneity of this expression pattern thus provides new insights into sterol metabolism and related disease treatment strategies. These findings demonstrate SRS-GEM as a promising platform for new possibilities in investigating metabolic regulation, disease mechanisms, and biopharmaceutical research.

Keywords: HMGCoA reductase; biophysics; cholesterol; genetic engineering; metabolite imaging; stimulated Raman scattering (SRS) microscopy; yeast.

Grants and funding

This work was supported by the National Natural Science Foundation of China (Research Fund for International Young Scientists, 32050410293), the Fundamental Research Funds for the Central Universities (2020QNA5027), and MOE Frontier Science Center for Brain Science and Brain-Machine Integration of Zhejiang University, Zhejiang University 100-Young Professor startup fund to HJL; the National Natural Science Foundation of China (General Program, 12074339) to DZ; and the National Natural Science Foundation of China (92057102) and a research fund from the Zhejiang Provincial Key Laboratory for Cancer Molecular Cell Biology to CY.