Immunofluorescence microscopy cytoskeleton

Small, J. V., Zobeley, S., Rinnerthaler, G., and Faulstich, H. (1988) Coumarin-phalloidin a new actin probe permitting triple immunofluorescence microscopy of the cytoskeleton. J. Cell Sci. 89,21-24. 

The structural integrity of microfilaments has been shown to be necessary for signal transduction within osteoblasts. Examination of the effects of mechanical strain on the expression of major structural elements of the cytoskeleton indicated that the amount of tubulin decreased by 75% and the amount of vinculin, a major component of focal adhesion complexes, increased by about 250%. Immunofluorescence microscopy demonstrated that mechanical strain led to increased formation and thickening of actin stress fibers, with dissociation of the microtubules and a clear increase in levels of vinculin at the peripheral edges of the cell. This suggests that mechanical strain leads to a coordinated change in both the cytoskeleton and in ECM proteins that facilitate tighter adhesion of osteoblasts and their ECM. 

The location of CD in the smooth muscle cell has been determined by immunofluorescence microscopy. A number of studies have shown the presence of two populations of thin filaments in smooth muscles one population is mainly p-actin and associated with the cytoskeleton, filamin, and calponin, whereas the remainder (about 60% of the total in chicken gizzard) is mainly a-actin and is associated with contractile proteins, myosin, TM, and CD (Furst et al., 1986 North et al., 1994a,b Lehman etal, 1987). 

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