Directional reorientation of migrating neutrophils is limited by suppression of receptor input signaling at the cell rear through myosin II activity

• Published in: Nature communications 2021-11, Vol.12 (1), p.6619-6619, Article 6619
• Main Authors: Hadjitheodorou, Amalia, Bell, George R. R., Ellett, Felix, Shastry, Shashank, Irimia, Daniel, Collins, Sean R., Theriot, Julie A.
• Format: Article
• Language: English
• Subjects: 13/62; 14; 14/33; 14/35; 14/63; 631/80/2373; 631/80/84/1372; 631/80/85; cdc42 GTP-Binding Protein - metabolism; Cdc42 protein; Cell migration; Cell Movement - physiology; Cell Polarity - physiology; Channels; Chemotaxis - physiology; HL-60 Cells; Humanities and Social Sciences; Humans; Leukocytes (neutrophilic); Localization; Microfluidics; multidisciplinary; Myosin; Myosin Light Chains - metabolism; Myosin Type II - metabolism; Neutrophils; Neutrophils - physiology; Phosphorylation; Polarity; Polarization; Receptor mechanisms; Receptors; Rho-associated kinase; rhoA GTP-Binding Protein - metabolism; RhoA protein; Science; Science (multidisciplinary)
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-26622-z

To migrate efficiently to target locations, cells must integrate receptor inputs while maintaining polarity: a distinct front that leads and a rear that follows. Here we investigate what is necessary to overwrite pre-existing front-rear polarity in neutrophil-like HL60 cells migrating inside straight microfluidic channels. Using subcellular optogenetic receptor activation, we show that receptor inputs can reorient weakly polarized cells, but the rear of strongly polarized cells is refractory to new inputs. Transient stimulation reveals a multi-step repolarization process, confirming that cell rear sensitivity to receptor input is the primary determinant of large-scale directional reversal. We demonstrate that the RhoA/ROCK/myosin II pathway limits the ability of receptor inputs to signal to Cdc42 and reorient migrating neutrophils. We discover that by tuning the phosphorylation of myosin regulatory light chain we can modulate the activity and localization of myosin II and thus the amenability of the cell rear to ‘listen’ to receptor inputs and respond to directional reprogramming. Neutrophils migrate with remarkably stable front-rear polarization. Using optogenetic receptor control to induce reversal of polarization in restrictive microfluidic channels, the authors find that myosin II promotes this stability by suppressing transmission of receptor inputs at the cell rear.