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Updated: Aug 8, 2026
Quantifying Synapses: an Immunocytochemistry-based Assay to Quantify Synapse Number
Published on: November 17, 2010
Cutting edge: quantitative imaging of raft accumulation in the immunological synapse
W Richard Burack1, Kyeong-Hee Lee, Amy D Holdorf
1Department of Pathology and Immunology, Washington University School of Medicine, Saint Louis, MO 63110, USA.
Insights
Lipid rafts accumulate in the central zone of the immunological synapse (C-SMAC). This accumulation is modest and driven by rearrangement, independent of CD28 signaling.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Lipid rafts are known to accumulate at the immunological synapse.
- The precise localization, extent, and origin of lipid raft recruitment remain undefined.
Purpose of the Study:
- To define the quantitative and spatial characteristics of lipid raft accumulation at the immunological synapse.
- To investigate the role of CD28 in lipid raft recruitment to the immunological synapse.
Main Methods:
- Quantitative analysis of lipid raft distribution within the immunological synapse.
- Assessment of lipid raft recruitment in CD28-deficient cells.
Main Results:
- Lipid rafts preferentially accumulate in the central supramolecular activation complex (C-SMAC).
- Quantitative analysis shows modest lipid raft recruitment to the C-SMAC, suggesting rearrangement from peripheral zones.
- Lipid raft accumulation is independent of the CD28/B7 co-stimulatory pathway.
Conclusions:
- Lipid raft accumulation at the immunological synapse is primarily a rearrangement process.
- CD28 signaling does not significantly influence lipid raft recruitment to the immunological synapse.
Abstract:
Although the accumulation of lipid rafts at the immunological synapse is now well accepted, the degree of the accumulation, the localization within the fine structure of the immunological synapse, and the region from which lipid rafts are recruited have not been defined. In this work we show that lipid rafts preferentially accumulate in the central zone of the immunological synapse, the central supramolecular activation complex (C-SMAC). However, quantitative analyses indicate that the level of recruitment of lipid rafts to the C-SMAC is relatively small and suggests that rearrangement of lipid rafts from the peripheral zone of the synapse into the C-SMAC can account for this accumulation. We also assessed the effects of CD28 deficiency on lipid raft recruitment to the immunological synapse. The accumulation of lipid occurred independently of the CD28/B7 system and was not measurably altered by CD28.
