2018
DOI: 10.1002/jbmr.3409
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Live Imaging of Type I Collagen Assembly Dynamics in Osteoblasts Stably Expressing GFP and mCherry-Tagged Collagen Constructs

Abstract: Type I collagen is the most abundant extracellular matrix protein in bone and other connective tissues and plays key roles in normal and pathological bone formation as well as in connective tissue disorders and fibrosis. Although much is known about the collagen biosynthetic pathway and its regulatory steps, the mechanisms by which it is assembled extracellularly are less clear. We have generated GFPtpz and mCherry-tagged collagen fusion constructs for live imaging of type I collagen assembly by replacing the … Show more

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Cited by 44 publications
(39 citation statements)
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“…Macrophages directly contribute to collagen deposition and scar formation. To address whether macrophages contribute directly to collagen deposition in the P7 and adult mouse heart post-MI, we utilised a transgenic mouse line that expresses a collagen-GFP fusion protein to fluorescently label type-1 collagen fibrils 36 . We initially characterised GFPtpz-collagen deposition in adult mice post-MI ( Supplementary Fig.…”
Section: Resultsmentioning
confidence: 99%
“…Macrophages directly contribute to collagen deposition and scar formation. To address whether macrophages contribute directly to collagen deposition in the P7 and adult mouse heart post-MI, we utilised a transgenic mouse line that expresses a collagen-GFP fusion protein to fluorescently label type-1 collagen fibrils 36 . We initially characterised GFPtpz-collagen deposition in adult mice post-MI ( Supplementary Fig.…”
Section: Resultsmentioning
confidence: 99%
“…A tool that might assist with work of this type is GFP-tagged collagen and mouse lines expressing this protein. 74 In vitro experiments are presently underway using live-cell imaging of fibroblasts derived from GFP-collagen expressing mice to observe patterns of collagen laid down by motile cells and determine whether αCT1 increases the disorganization of newly secreted ECM. A further limitation is that we do not understand the molecular bases of the effect of the peptide on dermal collagen order in vivo.…”
Section: Discussionmentioning
confidence: 99%
“…Hence, other methodologies should be adopted when dynamic events involved in bone formation, such as extracellular collagen assembly, are the imaging target. Lu et al revealed the dynamics of in vitro extracellular collagen assembly using time‐lapse confocal imaging. Imaging in cultured osteoblasts expressing fluorescent collagen and fibronectin‐null cells showed that collagen assembly is highly dynamic and influenced by constantly stretching and contracting from motile cells, and that collagen assembly is dependent on fibronectin deposition as a dynamically integrative event.…”
Section: Imaging Of Bone Matricesmentioning
confidence: 99%