The dynamics of secretion during sea urchin embryonic skeleton formation.
Skeleton formation involves secretion of massive amounts of mineral precursor, usually a calcium salt, and matrix proteins, many of which are deposited on, or even occluded within, the mineral. The cell biological underpinnings of this secretion and subsequent assembly of the biomineralized skeletal element is not well understood. We ask here what is the relationship of the trafficking and secretion of the mineral and matrix within the primary mesenchyme cells of the sea urchin embryo, cells that deposit the endoskeletal spicule. Fluorescent labeling of intracellular calcium deposits show mineral precursors are present in granules visible by light microscopy, from whence they are deposited in the endoskeletal spicule, especially at its tip. In contrast, two different matrix proteins tagged with GFP are present in smaller post-Golgi vesicles only seen by electron microscopy, and the secreted protein are only incorporated into the spicule in the vicinity of the cell of origin. The matrix protein, SpSM30B, is post-translationally modified during secretion, and this processing continues after its incorporation into the spicule. Our findings also indicate that the mineral precursor and two well characterized matrix proteins are trafficked by different cellular routes.
Wilt
Fred H
FH
Department of Molecular and Cell Biology, University of California, 142 Life Sciences Addition, Berkeley, CA 94720-3200, USA. wilt@socrates.berkeley.edu <wilt@socrates.berkeley.edu>
Killian
Christopher E
CE
Hamilton
Patricia
P
Croker
Lindsay
L
D000818:Animals; D002113:Calcification, Physiologic; D002118:Calcium / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000032:analysis; D047270:Strongylocentrotus purpuratus / Q000166:cytology / Q000196:embryology* / Q000378:metabolism
D003598:Cytoskeletal Proteins; D016326:Extracellular Matrix Proteins; D011993:Recombinant Fusion Proteins; C051691:SM50 protein, sea urchin; C071119:SM30 protein, Strongylocentrotus purpuratus; D049452:Green Fluorescent Proteins; D002118:Calcium