Abstract
Mouse keratinocytes migrate significantly slower than their human counterparts in vitro on uncoated surfaces. We tested the hypothesis that this is a consequence of differences in the extracellular matrix (ECM) that cells deposit. In support of this, human keratinocyte motility was markedly reduced when plated onto the ECM of mouse skin cells, whereas the latter cells migrated faster when plated onto human keratinocyte ECM. The ECM of mouse and human keratinocytes contained similar levels of the α3 laminin subunit of laminin-332. However, mouse skin cells expressed significantly more fibronectin (FN) than human cells. To assess whether FN is a motility regulator, we used small interfering RNA (siRNA) to reduce the expression of FN in mouse keratinocytes. The treated mouse keratinocytes moved significantly more rapidly than wild-type mouse skin cells. Moreover, the FN-depleted mouse cell ECM supported increased migration of both mouse and human keratinocytes. Furthermore, the motility of human keratinocytes was slowed when plated onto FN-coated substrates or human keratinocyte ECM supplemented with FN in a dose-dependent manner. Consistent with these findings, the ECM of α3 integrin-null keratinocytes, which also migrated faster than wild-type cells, was FN deficient. Our results provide evidence that FN is a brake to skin cell migration supported by laminin-332-rich matrices.
Original language | English (US) |
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Pages (from-to) | 448-457 |
Number of pages | 10 |
Journal | Journal of Investigative Dermatology |
Volume | 132 |
Issue number | 2 |
DOIs | |
State | Published - Feb 2012 |
Funding
We thank Lou Laimins of Northwestern University for providing immortalized human keratinocytes. A Dermatology Foundation Research grant and a Career Development Award support KH. Work in the Jones laboratory is funded by NIAMS (R01 AR054184) and in the Green laboratory by NIAMS (R01 AR041836). This research was supported in part by resources provided by the Northwestern University Skin Disease Research Center (5P30AR057216-02), Chicago, IL, with support from the NIH/NIAMS. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the Northwestern University Skin Disease Research Center or the NIH/NIAMS. Imaging work was performed at the Northwestern University Cell Imaging Facility generously supported by NCI CCSG P30 CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center. FACS analyses were performed at the Northwestern University Flow Cytometry Facility supported by a Cancer Center Support grant (NCI CA060553).
ASJC Scopus subject areas
- Dermatology
- Molecular Biology
- Biochemistry
- Cell Biology