An ultrastructural study of the microfilaments in rat brain by means of heavy meromyosin labeling. I. The perikaryon, the dendrites and the axon.

Abstract

Nervous tissue pieces from the caudate nucleus and the substantia nigra of the rat were incubated in cold glycerol solutions of decreasing concentrations and then transferred into standard phosphate buffer (pH 7.0) or into tris-K+-Mg++-Ca++ buffer (pH 7.9) containing HMM, prepared from rabbit skeletal muscle by tryptic digestion. As controls, pieces were immersed for an identical period in the same buffers (1) without HMM or (2) with HMM to which had been added 2.5 mM Na+ pyrophosphate or 5 mM ATP. In control neurons smooth-surfaced microfilaments, about 50 A in diameter, were observed. After reaction with HMM, the microfilaments were increased in number and density and in width to 180-200 A. A meshwork was formed. Arrowheads pointing in the same direction were spaced at regular intervals (300-350 A) among short segments of the surfaces of the microfilaments, depending upon the plane of section. More often, however, typical arrowheads were not observed, and the surfaces of the microfilaments were seen coated with polarized side-arms cross-bridging the spaces between adjacent elements at more or less regular intervals. When cross-sectioned, the microfilaments appeared as dense dots from which a material of lesser electron density radiated. Following incubation in HMM solutions containing Na+ pyrophosphate or ATP, no arrowhead structures were seen. Of particular interest was the structural relation of the actin-like filaments with occasional, tapered myosin-like filaments, and with the plasma membrane, which served as anchor points. Mitochondria and smooth ER membranes were observed to be attached to the actin-like filaments or enmeshed in the network. The microtubules, as well as most of the neurofilaments, were disrupted by the glycerination procedure at 4 degrees, and thus no precision about the structural relationship of the actin-like filaments with the latter elements could be added. The role of the actin-like filaments in the transport of material, by a mechanism of chemomechanical transduction, throughout the neuron from sites of synthesis to functional locations, and between several functional locations, is discussed.