Abstract
Chemotaxis, the directional cell migration guided by chemoattractant gradients, plays essential roles in many physiological processes, such as recruitment of neutrophils to sites of inflammation. Neutrophils detect chemoattractants by G protein-coupled receptors (GPCRs). Chemoattractant stimuli activate multiple signaling pathways to regulate directional migration of neutrophils. Recently, we identified a novel GPCR-mediated PLCβγ/ PKCβ/PKD1 signaling axis that regulates cofilin activity through cofilin phosphatase slingshot 2 (SSH2) and remodels actin cytoskeleton during neutrophil chemotaxis. In the future, it will be important to understand how multiple signaling pathways are spatiotemporally regulated to precisely control the rapid remodeling of actin cytoskeleton in the leading front of chemotaxing neutrophils.
Highlights
Chemotaxis, the directional cell migration guided by chemoattractant gradients, plays essential roles in many physiological processes, such as recruitment of neutrophils to sites of inflammation
The ability to screen and analyse large cell populations in a high-throughput manner is crucial in today’s research [4]. To evaluate this imaging technology, we evaluated cellular morphology using Amnis® ImageStreamX Mark II imaging flow cytometer and compared this with experienced human assessment
Circularity describes how ruffled or irregular the membranes of cells are while aspect ratio describes the overall shape of the cells in terms of how elongated cells are. 500 cells were obtained by pressing a commercially available membrane (Eyeprim, Opa Tech) on the conjunctiva of a human participant for 2-5 seconds
Summary
Chemotaxis, the directional cell migration guided by chemoattractant gradients, plays essential roles in many physiological processes, such as recruitment of neutrophils to sites of inflammation. One recent type of automated cellular imaging technology, the Amnis, combines the capabilities of microscopy and flow cytometry in a single platform and is able to perform cellular imaging and automatic calculation of quantitative cellular morphological indices [1,2,3]. We validated 2 common imaging parameters, circularity (Figure 1A) and aspect ratio (Figure 1D).
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