Actin filaments are required for fibripositor-mediated collagen fibril alignment in tendon.
Cells in tendon deposit parallel arrays of collagen fibrils to form a functional tissue, but how this is achieved is unknown. The cellular mechanism is thought to involve the formation of intracellular collagen fibrils within Golgi to plasma membrane carriers. This is facilitated by the intracellular processing of procollagen to collagen by members of the tolloid and ADAMTS families of enzymes. The carriers subsequently connect to the extracellular matrix via finger-like projections of the plasma membrane, known as fibripositors. In this study we have shown, using three-dimensional electron microscopy, the alignment of fibripositors with intracellular fibrils as well as an orientated cable of actin filaments lining the cytosolic face of a fibripositor. To demonstrate a specific role for the cytoskeleton in coordinating extracellular matrix assembly, cytochalasin was used to disassemble actin filaments and nocodazole or colchicine were used to disrupt microtubules. Microtubule disruption delayed procollagen transport through the secretory pathway, but fibripositor numbers were unaffected. Actin filament disassembly resulted in rapid loss of fibripositors and a subsequent disappearance of intracellular fibrils. Procollagen secretion or processing was not affected by cytochalasin treatment, but the parallelism of extracellular collagen fibrils was altered. In this case a significant proportion of collagen fibrils were found to no longer be orientated with the long axis of the tendon. The results suggest an important role for the actin cytoskeleton in the alignment and organization of the collagenous extracellular matrix in embryonic tendon.
Canty
Elizabeth G
EG
Wellcome Trust Centre for Cell Matrix Research, Faculty of Life Sciences, University of Manchester, Michael Smith Building, Oxford Road, Manchester M13 9PT, United Kingdom.
Starborg
Tobias
T
Lu
Yinhui
Y
Humphries
Sally M
SM
Holmes
David F
DF
Meadows
Roger S
RS
Huffman
Adam
A
O'Toole
Eileen T
ET
Kadler
Karl E
KE
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002642:Chick Embryo; D003094:Collagen / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D013710:Tendons / Q000166:cytology / Q000196:embryology / Q000201:enzymology / Q000378:metabolism*
D000199:Actins; D003094:Collagen