High-resolution imaging and quantification of plasma membrane cholesterol by NanoSIMS

He, Cuiwen; Hu, Xuchen; Jung, Rachel S.; Weston, Thomas A.; Sandoval, Norma P.; Tontonoz, Peter; Kilburn, Matt R.; Fong, Loren G.; Young, Stephen G.; Jiang, Haibo

doi:10.1073/pnas.1621432114

Cited by 74 publications

(63 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Because the particles originate from the plasma membrane, we suspected that they would contain sphingomyelin. To test this idea, we incubated cholesterol-loaded RAW 264.7 macrophages and primary macrophages with [ 15 N]ALO-D4 and [ 13 C]lysenin [a sphingomyelin-binding cytolysin (15)]. 15 N/ 14 N and 13 C/ 12 C nanoSIMS images revealed that macrophage-derived particles were enriched in both 15 N and 13 C ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…A linear regression analysis revealed a positive correlation between 15 N and 13 C enrichments (P < 0.0001). 15 N/ 14 N and 13 C/ 12 C scales are multiplied by 10,000. Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The accessible pool of cholesterol on the plasma membrane is metabolically important because it is mobile, capable of moving to the endoplasmic reticulum and participating in the regulation of cholesterol biosynthetic enzymes (14). Recently, He et al (15) used nanoSIMS, along with an 15 N-labeled ALO-D4, to show that the accessible pool of cholesterol on the plasma membrane of CHO-K1 cells is concentrated on microvilli.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Macrophages release plasma membrane-derived particles rich in accessible cholesterol

Weston

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

View full text Add to dashboard Cite

Macrophages are generally assumed to unload surplus cholesterol through direct interactions between ABC transporters on the plasma membrane and HDLs, but they have also been reported to release cholesterol-containing particles. How macrophage-derived particles are formed and released has not been clear. To understand the genesis of macrophage-derived particles, we imaged mouse macrophages by EM and nanoscale secondary ion mass spectrometry (nanoSIMS). By scanning EM, we found that large numbers of 20- to 120-nm particles are released from the fingerlike projections (filopodia) of macrophages. These particles attach to the substrate, forming a "lawn" of particles surrounding macrophages. By nanoSIMS imaging we showed that these particles are enriched in the mobile and metabolically active accessible pool of cholesterol (detectable by ALO-D4, a modified version of a cholesterol-binding cytolysin). The cholesterol content of macrophage-derived particles was increased by loading the cells with cholesterol or by adding LXR and RXR agonists to the cell-culture medium. Incubating macrophages with HDL reduced the cholesterol content of macrophage-derived particles. We propose that release of accessible cholesterol-rich particles from the macrophage plasma membrane could assist in disposing of surplus cholesterol and increase the efficiency of cholesterol movement to HDL.

show abstract

Section: Resultsmentioning

confidence: 99%

“…A linear regression analysis revealed a positive correlation between 15 N and 13 C enrichments (P < 0.0001). 15 N/ 14 N and 13 C/ 12 C scales are multiplied by 10,000. Fig.…”

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Macrophages release plasma membrane-derived particles rich in accessible cholesterol

Weston

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Tissue sections were coated with 5 nm of platinum and analyzed with NanoSIMS 50L or NanoSIMS 50 instruments [7, 8]. A 16-KeV 133 Cs + beam was used to bombard the sample, and secondary electrons (SEs) and secondary ions ( e.g., 12 C 14 N–, 1 H–, 2 H–, 12 C–, 13 C–) were collected [7].…”

Section: Methodsmentioning

confidence: 99%

NanoSIMS imaging reveals unexpected heterogeneity in nutrient uptake by brown adipocytes

Weston

et al. 2018

Biochemical and Biophysical Research Communications

Self Cite

View full text Add to dashboard Cite

Heterogeneity in the metabolic properties of adipocytes in white adipose tissue has been well documented. We sought to investigate metabolic heterogeneity in adipocytes of brown adipose tissue (BAT), focusing on heterogeneity in nutrient uptake. To explore the possibility of metabolic heterogeneity in brown adipocytes, we used nanoscale secondary ion mass spectrometry (NanoSIMS) to quantify uptake of lipids in adipocytes interscapular BAT and perivascular adipose tissue (PVAT) after an intravenous injection of triglyceride-rich lipoproteins (TRLs) containing [H]triglycerides (H-TRLs). The uptake of deuterated lipids into brown adipocytes was quantified by NanoSIMS. We also examined C enrichment in brown adipocytes after administering [C]glucose or C-labeled mixed fatty acids by gastric gavage. The uptake ofH-TRLs-derived lipids into brown adipocytes was heterogeneous, with H enrichment in adjacent adipocytes varying by more than fourfold. We also observed substantial heterogeneity inC enrichment in adjacent brown adipocytes after administering [C]glucose or [C]fatty acids by gastric gavage. The uptake of nutrients by adjacent brown adipocytes within a single depot is variable, suggesting that there is heterogeneity in the metabolic properties of brown adipocytes.

show abstract

“…8 The accumulation of sphingomyelin in the microvilli was recently confirmed by using nanoscale secondary ion mass spectrometry (NanoSIMS) imaging. 9 Kai Simons and colleagues also conducted a detailed study on the lipid composition of apical membrane by using mass spectrometry. 10 They reported that sphingomyelin is enriched in the apical membrane, compared to the whole cell lipids, by physically isolating apical membrane using nitrocellulose membrane.…”

Section: Roles Of Sphingolipids In the Formation Of Apical Membranementioning

confidence: 99%

Roles of membrane lipids in the organization of epithelial cells: Old and new problems

Ikenouchi

2018

Tissue Barriers

View full text Add to dashboard Cite

Epithelial cells have characteristic membrane domains. Identification of membrane proteins playing an important role in these membrane domains has progressed and numerous studies have been performed on the functional analysis of these membrane proteins. On the other hand, the precise roles of membrane lipids in the organization of these membrane domains are largely unknown. Historically, the concept of lipid raft arose from the analysis of lipid composition of the apical membrane, and it can be said that epithelial cells are an optimal experimental model for elucidating the functions of lipids. In this review, I discuss the role of lipids in the formation of epithelial polarity and in the formation of cell membrane structures of epithelial cells such as microvilli in the apical domain, cell-cell adhesion apparatus in the lateral domain and cell-matrix adhesion in the basal domain.

show abstract

High-resolution imaging and quantification of plasma membrane cholesterol by NanoSIMS

Cited by 74 publications

References 16 publications

Macrophages release plasma membrane-derived particles rich in accessible cholesterol

Macrophages release plasma membrane-derived particles rich in accessible cholesterol

NanoSIMS imaging reveals unexpected heterogeneity in nutrient uptake by brown adipocytes

Roles of membrane lipids in the organization of epithelial cells: Old and new problems

Contact Info

Product

Resources

About