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Primary fibroblasts isolated from foetal mouse cornea, skin and tendon were subjected to linear shear stress and analysed for morphological parameters and by microarray, as compared with unstimulated controls. Approx. 350 genes were either up- or down-regulated by a significant amount, with 51 of these being common to all three cell types. Approx. 50% of altered genes in tendon and cornea fibroblasts were changed in common with one of the other cell types, with the remaining approx. 50% being specific to tendon or cornea. In skin fibroblasts, however, less than 25% of genes whose transcription was altered were specific only to skin. The functional spectrum of genes that were up- or down-regulated was diverse, with apparent house-keeping genes forming the major category of up-regulated genes. However, a significant number of genes associated with cell adhesion, extracellular matrix and matrix remodelling, as well as cytokines and other signalling factors, were also affected. Somewhat surprisingly, in these latter categories the trend was towards a reduction in mRNA levels. Verification of the mRNA quantity of a subset of these genes was performed by reverse transcriptase PCR and was found to be in agreement with the microarray analysis. These findings provide the first in-depth analysis of phenotypic differences between fibroblast cells from different tissue sources and reveal the responses of these cells to mechanical stress.

The sodium-hydrogen exchanger isoform-1 (NHE1) functions in intracellular pH and cell volume homeostasis by catalyzing an electroneutral exchange of extracellular sodium and intracellular hydrogen. Recent studies have revealed the structural functions of NHE1 as an anchor for actin filaments and a scaffold for an ensemble of signaling proteins. This review highlights how these functions contribute to NHE1 regulation of biochemical events and cell behaviors.New data confirming nontransport structural functions of NHE1 suggest reexamining how NHE1 regulates cell functions. Cell survival, cell substrate adhesion, and organization of the actin cytoskeleton are confirmed to be regulated through actin anchoring by NHE1 and likely by NHE1-dependent scaffolding of signaling proteins. A role for NHE1 in mechanotransduction is emerging and a challenge of future studies is to determine whether structural functions of NHE1 are important for mechanoresponsiveness.This review highlights evidence for the nontransport functions of NHE1 and describes how the structural functions are integrated with ion translocation to regulate a range of cellular processes. Nontransporting features of NHE1 are analogous to recently observed nonconducting actions of ion channels in regulating cell behaviors and represent an emerging paradigm of ion transporters as multifunctional proteins.

Cellular stress activates transcription of various genes that mediate stress‐driven proliferation and differentiation in many cells including osteoblasts, endothelial cells, and fibroblasts. In response to mechanical stress, expression of some genes is altered regardless of cell types and that of others in specific cell types. Using the microarray‐based expression data for primary fibroblasts isolated from fetal mouse cornea, skin and tendon, we conducted a model‐based transcription analysis and predicted transcription‐factor binding motifs (TFBMs) responsible for the observed gene alteration. The computational procedure was formulated as a combinatorial optimization problem, and the AntModeler using an ant algorithm was employed to select TFBMs for each of the three fibroblast types. The results indicate that the stress responses are regulated mostly through cell type specific TFBMs together with a limited number of common TFBMs. The predicted role of those TFBMs should be evaluated experimentally.