Axonal growth in response to experimentally applied mechanical tension

Abstract

A machine was constructed, called a Cell Puller, that allows the steady advance or withdrawal of a microelectrode at very slow speeds-up to 170 microns/hr. Specially prepared microelectrodes held in the Cell Puller were placed in cultures of dissociated chick sensory ganglion neurons in such a way that growth cones attached to their tips. Movements of the microelectrodes, at speeds up to about 100 microns/hr, then resulted in the elongation of the neurites for up to 24 hr and for increases in length up to 960 microns; more rapid towing failed to cause extensive neurite elongation. Estimates of neurite diameter before and after "towing" indicated that a net increase in neurite volume had occurred. Furthermore, long neurites could be produced by towing from previously rounded neuronal cell bodies confined to small adhesive "islands" on a nonadhesive substratum. Neurites produced by microelectrode towing had a normal appearance, showed rapid saltatory movements of internal organelles and were capable of resuming growth on the substratum. Electron microscopy of bundles of neurites produced in this way from explanted dorsal root ganglia showed an ultrastructure typical of cultured neurites, with abundant longitudinally aligned microtubules and neurofilaments. These experiments demonstrate that neurites can grow in response to mechanical tension under tissue culture conditions. It is proposed that they do so also in normal development, the tension in this case being supplied initially by the locomotory activity of the growth cones and subsequently by the morphogenetic movements of the surrounding tissues.