The secreted metalloprotease ADAMTS20 is required for melanoblast survival.
ADAMTS20 (Adisintegrin-like and metalloprotease domain with thrombospondin type-1 motifs) is a member of a family of secreted metalloproteases that can process a variety of extracellular matrix (ECM) components and secreted molecules. Adamts20 mutations in belted (bt) mice cause white spotting of the dorsal and ventral torso, indicative of defective neural crest (NC)-derived melanoblast development. The expression pattern of Adamts20 in dermal mesenchymal cells adjacent to migrating melanoblasts led us to initially propose that Adamts20 regulated melanoblast migration. However, using a Dct-LacZ transgene to track melanoblast development, we determined that melanoblasts were distributed normally in whole mount E12.5 bt/bt embryos, but were specifically reduced in the trunk of E13.5 bt/bt embryos due to a seven-fold higher rate of apoptosis. The melanoblast defect was exacerbated in newborn skin and embryos from bt/bt animals that were also haploinsufficient for Adamts9, a close homolog of Adamts20, indicating that these metalloproteases functionally overlap in melanoblast development. We identified two potential mechanisms by which Adamts20 may regulate melanoblast survival. First, skin explant cultures demonstrated that Adamts20 was required for melanoblasts to respond to soluble Kit ligand (sKitl). In support of this requirement, bt/bt;Kit(tm1Alf)/+ and bt/bt;Kitl(Sl)/+ mice exhibited synergistically increased spotting. Second, ADAMTS20 cleaved the aggregating proteoglycan versican in vitro and was necessary for versican processing in vivo, raising the possibility that versican can participate in melanoblast development. These findings reveal previously unrecognized roles for Adamts proteases in cell survival and in mediating Kit signaling during melanoblast colonization of the skin. Our results have implications not only for understanding mechanisms of NC-derived melanoblast development but also provide insights on novel biological functions of secreted metalloproteases.
Silver
Debra L
DL
Genetic Disease Research Branch, National Human Genome Research Institute, Bethesda, Maryland, United States of America.
Hou
Ling
L
Somerville
Robert
R
Young
Mary E
ME
Apte
Suneel S
SS
Pavan
William J
WJ
D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism*; D000071096:ADAMTS Proteins; D000483:Alleles; D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival / Q000235:genetics / Q000502:physiology; D017931:DNA Primers / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D005260:Female; D006801:Humans; D008544:Melanocytes / Q000166:cytology* / Q000201:enzymology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D008822:Mice, Transgenic; D009154:Mutation; D009432:Neural Crest / Q000166:cytology / Q000196:embryology / Q000201:enzymology; D010641:Phenotype; D011247:Pregnancy; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D015398:Signal Transduction; D012867:Skin / Q000166:cytology / Q000196:embryology / Q000201:enzymology; D012880:Skin Pigmentation / Q000235:genetics / Q000502:physiology; D053675:Versicans / Q000378:metabolism
D017931:DNA Primers; C508323:Vcan protein, mouse; D053675:Versicans; D019009:Proto-Oncogene Proteins c-kit; D051722:ADAM Proteins; D000071096:ADAMTS Proteins; C479569:Adamts20 protein, mouse