Neoformation of intercellular adherens junctions accompanies the differentiation of fibroblasts into contractile myofibroblasts, a key event during development of fibrosis and in wound healing. We have previously shown that intercellular mechanical coupling of stress fibres via adherens junctions improves contraction of collagen gels by myofibroblasts. By assessing spontaneous intracellular Ca2+ oscillations, we here test whether adherens junctions mechanically coordinate myofibroblast activities. Periodic Ca2+ oscillations are synchronised between physically contacting myofibroblasts and become desynchronised upon dissociation of adherens junctions with function-blocking peptides. Similar uncoupling is obtained by inhibiting myofibroblast contraction using myosin inhibitors and by blocking mechanosensitive ion channels using Gd3+ and GSMTx4. By contrast, gap junction uncouplers do not affect myofibroblast coordination. We propose the following model of mechanical coupling for myofibroblasts: individual cell contraction is transmitted via adherens junctions and leads to the opening of mechanosensitive ion channels in adjacent cells. The resulting Ca2+ influx induces a contraction that can feed back on the first cell and/or stimulate other contacting cells. This mechanism could improve the remodelling of cell-dense tissue by coordinating the activity of myofibroblasts.
A new model based on ray tracing was developed to estimate power spectral properties in laser Doppler velocimetry of retinal vessels and to predict the effects of laser beam size and eccentricity as well as absorption of laser light by oxygenated and reduced hemoglobin. We describe the model and show that it correctly converges to the traditional rectangular shape of the Doppler shift power spectrum, given the same assumptions, and that reduced beam size and eccentric alignment cause marked alterations in this shape. The changes in the detected total power of the Doppler-shifted light due to light scattering and absorption by blood can also be quantified with this model and may be used to determine the oxygen saturation in retinal arteries and veins. The potential of this approach is that it uses direct measurements of Doppler signals originating from moving red blood cells. This may open new avenues for retinal vessel oximetry.
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