Abstract
SummaryThe five‐subunit WASH complex generates actin networks that participate in endocytic trafficking, migration and invasion in various cell types. Loss of one of the two subunits WASH or strumpellin in mice is lethal, but little is known about their role in mammals in vivo. We explored the role of strumpellin, which has previously been linked to hereditary spastic paraplegia, in the mouse melanocytic lineage. Strumpellin knockout in melanocytes revealed abnormal endocytic vesicle morphology but no impairment of migration in vitro or in vivo and no change in coat colour. Unexpectedly, WASH and filamentous actin could still localize to vesicles in the absence of strumpellin, although the shape and size of vesicles was altered. Blue native PAGE revealed the presence of two distinct WASH complexes, even in strumpellin knockout cells, revealing that the WASH complex can assemble and localize to endocytic compartments in cells in the absence of strumpellin.
Highlights
In the adult mouse, mature melanocytes localize to the base of hair follicles where they generate melanin to regulate coat colour
The five-subunit WASH complex generates actin networks that participate in endocytic trafficking, migration and invasion in various cell types
We explored the role of strumpellin, which has previously been linked to hereditary spastic paraplegia, in the mouse melanocytic lineage
Summary
Mature melanocytes localize to the base of hair follicles where they generate melanin to regulate coat colour. Melanoblasts originate at the neural crest at ~E8.5, and a subset migrate on a dorsal–lateral pathway to populate the skin of the whole embryo by ~E15.5 and home to hair follicles. Another population of melanoblasts may arise from Schwann cell precursors along peripheral nerves underneath the skin (Adameyko and Lallemend, 2010; Adameyko et al, 2009). Melanoblasts migrate in the epidermis with long protrusions in a Rac1-dependent manner, using actin assembly to drive elongation of the cell and translocation between keratinocytes (Li et al, 2011). It is unknown whether melanoblasts depend on integrin and receptor recycling via the activity of the WASH complex for their migration, as has been reported for other cell types in a 3D environment (Dozynkiewicz et al, 2012; Muller et al, 2009; Zech et al, 2011)
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