TY - JOUR
T1 - ALIX and ceramide differentially control polarized small extracellular vesicle release from epithelial cells
AU - Matsui, Takahide
AU - Osaki, Futaba
AU - Hiragi, Shu
AU - Sakamaki, Yuriko
AU - Fukuda, Mitsunori
N1 - Funding Information:
We thank K. Shoji for technical assistance, and E. Morita, N. Tanaka, and all members of the Fukuda laboratory for helpful discussions. This work was supported in part by Grant‐in‐Aid for Research Activity Start‐up 19K21174 from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan (to TM); the Kao Foundation for Arts and Sciences (to TM); Grant‐in‐Aid for Scientific Research (B) 19H03220 from MEXT (to MF); and Japan Science and Technology Agency (JST) CREST Grant JPMJCR17H4 (to MF).
Publisher Copyright:
© 2021 The Authors
PY - 2021/5/5
Y1 - 2021/5/5
N2 - Exosomes, important players in cell–cell communication, are small extracellular vesicles of endocytic origin. Although single cells are known to release various kinds of exosomes (referred to as exosomal heterogeneity), very little is known about the mechanisms by which they are produced and released. Here, we established methods of studying exosomal heterogeneity by using polarized epithelial cells and showed that distinct types of small extracellular vesicles (more specifically CD9- and CD63-positive, Annexin I-negative small extracellular vesicles, which we refer to as exosomes herein) are differentially secreted from the apical and basolateral sides of polarized epithelial cells. We also identify GPRC5C (G protein-coupled receptor class C group 5 member C) as an apical exosome-specific protein. We further demonstrate that basolateral exosome release depends on ceramide, whereas ALIX, an ESCRT (endosomal sorting complexes required for transport)-related protein, not the ESCRT machinery itself, is required for apical exosome release. Thus, two independent machineries, the ALIX–Syntenin1–Syndecan1 machinery (apical side) and the sphingomyelinase-dependent ceramide production machinery (basolateral side), are likely to be responsible for the polarized exosome release from epithelial cells.
AB - Exosomes, important players in cell–cell communication, are small extracellular vesicles of endocytic origin. Although single cells are known to release various kinds of exosomes (referred to as exosomal heterogeneity), very little is known about the mechanisms by which they are produced and released. Here, we established methods of studying exosomal heterogeneity by using polarized epithelial cells and showed that distinct types of small extracellular vesicles (more specifically CD9- and CD63-positive, Annexin I-negative small extracellular vesicles, which we refer to as exosomes herein) are differentially secreted from the apical and basolateral sides of polarized epithelial cells. We also identify GPRC5C (G protein-coupled receptor class C group 5 member C) as an apical exosome-specific protein. We further demonstrate that basolateral exosome release depends on ceramide, whereas ALIX, an ESCRT (endosomal sorting complexes required for transport)-related protein, not the ESCRT machinery itself, is required for apical exosome release. Thus, two independent machineries, the ALIX–Syntenin1–Syndecan1 machinery (apical side) and the sphingomyelinase-dependent ceramide production machinery (basolateral side), are likely to be responsible for the polarized exosome release from epithelial cells.
KW - ESCRT
KW - ceramide
KW - exosome
KW - multivesicular body
KW - small extracellular vesicle
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U2 - 10.15252/embr.202051475
DO - 10.15252/embr.202051475
M3 - Article
C2 - 33724661
AN - SCOPUS:85102617825
VL - 22
JO - EMBO Reports
JF - EMBO Reports
SN - 1469-221X
IS - 5
M1 - e51475
ER -