Unique geometry of actin-membrane anchorage sites in avian gizzard smooth muscle cells

Abstract

Adherens junctions in isolated avian gizzard smooth muscle cells appear as short longitudinal streaks or chevrons that are arranged in periodic, mainly transverse bands along the cell surface. This barrel-like geometry, revealed by antibodies to either vinculin or talin, was seen also in teased gizzard strips by confocal laser-scanning microscopy and contrasted with the rib-like surface pattern observed here and previously in other avian and mammalian smooth muscles. There were on average 67 transverse bands per gizzard cell and an estimated total of around 800 vinculin/talin sites. The longitudinal spacing between the transverse bands of vinculin streaks in the gizzard cells changed from 4–5 microns in extended cells to around 1 micron in shortened cells and the bands remained essentially transverse at all cell lengths, inconsistent with a screw-like mode of cell shortening as has been invoked for smooth muscle cells by others. The absence of rotation on shortening was confirmed by observations on isolated and bead-decorated skinned cells that were induced to contract with ATP. Counterlabelling of cells with alpha-actinin antibodies produced more or less exclusive staining of the cytoplasmic dense bodies, and little surface label: the total number of dense bodies per cell, estimated from confocal microscope through focal series was in the range of 3000. The data are consistent with a periodic anchorage of actin filaments to the cell surface and, in turn, with the existence of regularly spaced contractile assemblies.