This study investigates the intercellular communication between neurons and oligodendrocytes through the release of exosomes. By utilizing a contact-free co-culture system, the research explores the mechanisms of calcium signaling and exosome secretion.
Begin with a contact-free co-culture of neurons and oligodendrocytes.
A permeable membrane separates the neurons in the transwell insert from the oligodendrocytes at the bottom well.
Add potassium to the insert to raise extracellular potassium levels, which inhibits the efflux of intracellular potassium.
The potassium accumulation depolarizes the neurons, triggering the opening of voltage-gated calcium channels and allowing calcium influx.
The influx stimulates the release of excitatory neurotransmitters, which diffuse across the permeable membrane to reach the bottom well and activate receptors on oligodendrocytes, leading to calcium influx.
The rise in calcium activates signaling pathways that induce the inward budding of membranes in early endosomes.
The budding encloses cytoplasmic proteins and RNAs within vesicles called exosomes inside multivesicular bodies or MVBs.
The MVBs fuse with the oligodendrocyte plasma membrane, releasing exosomes into the extracellular environment.
The secreted exosomes are now ready to assess intercellular communication.
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