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A novel function of LAR clustering in cytoneme-like membrane protrusions formation

Nguyen, Quynh Mai 大阪大学 DOI:10.18910/82350

2021.03.24

概要

Intercellular communication between cells distally located from each other can be assured by the formation of membrane protrusions that elongate from one cell to its target and function as conduits for the trafficking of various signaling factors. One type of the membrane protrusions called “cytonemes”- defined as thin, close ended protrusions enriched in actin filaments — is receiving more attentions in regards to its formation in many types of cells supporting numerous events like- establishment of the morphogen gradients, pattern formation, directed morphogenesis, and maintenance of the stem cell niche. Despite their crucial roles in cell signaling, the molecular mechanism by which cytonemes are established and maintained, however, remains elusive. In this study, I showed that the leukocyte common antigen-related (LAR) protein tyrosine phosphatase receptor plays an important role in cytoneme-like membrane protrusions formation. Exogenous expression of LAR in the human derived cell line induced formation of actin rich thin membrane protrusions, some of which exceed lengths of 200 gm and displayed a complex morphology with branches. To address how LAR functions in the cytoneme-like protrusions formation, I focused on the regulation of LAR dimerization/oligomerization by the extracellular matrix components- heparan sulphates (HS), chondroitin sulphates (CS) and the resulting regulatory effects on LAR phosphatase activity. Longer and more branched LAR-induced cytoneme-like membrane protrusions were observed with HS application compared to vehicle or CS treatments. These results were supported with the fact that a newly established rapamycin-induced LAR dimerization system also displayed significant increase in the formed longest protrusion length and complexity. Interestingly, although the truncated form of LAR entirely lacking the phosphatase-related domains DI and D2 (LAR_AD1D2)resulted into promotion of cytoneme-like protrusions formation, the phosphatase-inactive forms did not show clear changes suggesting that LAR clustering triggers formation of a molecular assembly that activates the formation of cytoneme-like protrusions in a phosphatase-independent manner. Altogether with the fact that axon (a well- known type of membrane protrusion formed in neurons) and cytonemes formation machineries share common cytoskeletal regulating factors like Arp2/3, Ena/VASP, Rael, the current study results hint on the existence of a common molecular mechanism for membrane protrusions establishment in the cell-to-cell contact formation, the one in which LAR is a key player.

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