Chemically skinned rabbit psoas muscle fibers/bundles were osmotically compressed with a macromolecule dextran T-500 (0-16%, g/100 ml) at 20 degrees C, 200 mM ionic strength, and pH 7.0. The lattice spacing of psoas bundles was measured by equatorial x-ray diffraction studies during relaxation and after rigor induction, and the results were compared with the fiber width measurements by optical microscopy. The purpose of the present study is to determine whether fiber width is a reliable measure of the lattice spacing, and to determine the available spacing for myosin cross-bridges between the thick and thin filaments. We observed that both the lattice spacing and the fiber width decreased with an increase in the dextran concentration during relaxation or after rigor induction, and that the spacing and the fiber width were proportionately related. We further observed that, in the absence of dextran, the lattice spacing (and the fiber width) shrank on a relax-to-rigor transition, whereas in the presence of 16% dextran, the spacing expanded on a relax-to-rigor transition. The cross-over of these plots occurred at the 4-7% dextran concentration. During Ca activation, the fiber width shrank in the absence of dextran, and it slightly expanded in the presence of 14.4% dextran. The degree of expansion was not as large as in the relax-to-rigor transition, and the cross-over occurred at about 11% dextran concentration. We also carried out experiments with dextran T-40 and T-10 to determine the upper limit of the molecular weight that enters the lattice space. We found that the upper limit is about 20 kD.