PMID- 37048046
OWN - NLM
STAT- MEDLINE
DCOM- 20230414
LR  - 20230427
IS  - 2073-4409 (Electronic)
IS  - 2073-4409 (Linking)
VI  - 12
IP  - 7
DP  - 2023 Mar 23
TI  - Characterization of Non-Cholesterol Sterols in Microglia Cell Membranes Using 
      Targeted Mass Spectrometry.
LID - 10.3390/cells12070974 [doi]
LID - 974
AB  - BACKGROUND: Non-cholesterol sterols, as well as plant sterols, cross the 
      blood-brain barrier and, thus, can be incorporated into cell membranes, affecting 
      the cell's inflammatory response. The aim of our work was to develop an 
      analytical protocol for a quantitative assessment of the sterol composition 
      within the membrane microdomains of microglia. METHODS: A protocol for cell 
      membrane isolation using OptiPrep(TM) gradient ultracentrifugation, in 
      combination with a targeted mass spectrometry (LC-MS/MS)-based assay, was 
      developed and validated for the quantitative analysis of free sterols in 
      microglia cell membranes. RESULTS: Utilizing an established LC-MS/MS assay, 
      cholesterol and seven non-cholesterol sterols were analyzed with a limit of 
      detection from 0.001 to 0.05 mg/L. Applying the detergent-free isolation of 
      SIM-A9 microglia cell membranes, cholesterol (CH), desmosterol (DE), lanosterol 
      (LA) stigmasterol (ST), beta-sitosterol (SI) and campesterol (CA) were quantified 
      with coefficients of variations between 6 and 29% (fractions 4-6, n = 5). The 
      highest concentrations of non-CH sterols within the microglia plasma membranes 
      were found in the microdomain region (DE>LA>SI>ST>CA), with ratios to CH ranging 
      from 2.3 to 435 lower abundancies. CONCLUSION: By applying our newly developed 
      and validated analytical protocol, we show that the non-CH sterol concentration 
      is about 38% of the total sterol content in microglia membrane microdomains. 
      Further investigations must clarify how changes in the non-sterol composition 
      influence membrane fluidity and cell signaling.
FAU - Begcevic Brkovic, Ilijana
AU  - Begcevic Brkovic I
AD  - Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, 
      University of Leipzig Medical Center, 04103 Leipzig, Germany.
FAU - Reinicke, Madlen
AU  - Reinicke M
AUID- ORCID: 0000-0003-3864-6424
AD  - Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, 
      University of Leipzig Medical Center, 04103 Leipzig, Germany.
FAU - Chey, Soroth
AU  - Chey S
AUID- ORCID: 0000-0001-7150-1791
AD  - Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, 
      University of Leipzig Medical Center, 04103 Leipzig, Germany.
FAU - Bechmann, Ingo
AU  - Bechmann I
AD  - Institute of Anatomy, Faculty of Medicine, University of Leipzig, 04103 Leipzig, 
      Germany.
FAU - Ceglarek, Uta
AU  - Ceglarek U
AD  - Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, 
      University of Leipzig Medical Center, 04103 Leipzig, Germany.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20230323
PL  - Switzerland
TA  - Cells
JT  - Cells
JID - 101600052
RN  - 0 (Sterols)
RN  - 0 (Phytosterols)
RN  - 99WUK5D0Y8 (Stigmasterol)
RN  - 1J05Z83K3M (Lanosterol)
SB  - IM
MH  - *Sterols/metabolism
MH  - Chromatography, Liquid
MH  - Microglia/metabolism
MH  - Tandem Mass Spectrometry
MH  - *Phytosterols
MH  - Stigmasterol
MH  - Lanosterol
MH  - Cell Membrane/metabolism
PMC - PMC10093698
OTO - NOTNLM
OT  - cholesterol precursors
OT  - microglia
OT  - plant sterols
OT  - plasma membrane microdomains
OT  - sterols
OT  - targeted mass spectrometry
COIS- The authors declare no conflict of interest. The funders had no role in the 
      design of the study; in the collection, analyses, or interpretation of data; in 
      the writing of the manuscript; or in the decision to publish the results.
EDAT- 2023/04/14 06:00
MHDA- 2023/04/14 06:42
PMCR- 2023/03/23
CRDT- 2023/04/13 01:16
PHST- 2022/12/09 00:00 [received]
PHST- 2023/03/09 00:00 [revised]
PHST- 2023/03/20 00:00 [accepted]
PHST- 2023/04/14 06:42 [medline]
PHST- 2023/04/13 01:16 [entrez]
PHST- 2023/04/14 06:00 [pubmed]
PHST- 2023/03/23 00:00 [pmc-release]
AID - cells12070974 [pii]
AID - cells-12-00974 [pii]
AID - 10.3390/cells12070974 [doi]
PST - epublish
SO  - Cells. 2023 Mar 23;12(7):974. doi: 10.3390/cells12070974.