OSMOTIC DILUTION STIMULATES AXONAL OUTGROWTH BY MAKING AXONS MORE SENSITIVE TO TENSION

Mechanical tension is a potent stimulator of axonal growth rate, which is also stimulated by osmotic dilution. We wished to determine the re lationship, if any, between osmotic stimulation and tensile regulation of axonal growth. We used calibrated glass needles to apply constant force to elongate axons of cultured chick sensory neurons. We find tha t a neurite being pulled at a constant force will grow 50-300% faster following a 50% dilution of inorganic ions in the culture medium. That is, osmotic dilution appears to cause axons to increase their sensiti vity to applied tensions. Experimental interventions suggest that this effect is not mediated by dilution of extracellular calcium, or to os motic stimulation of adenylate cyclase, or to osmotic stimulation of m echanosensitive ion channels. Rather, experiments measuring the static tension normally borne by neurites suggest a direct mechanical effect on the cytoskeletal proteins of the neurite shaft. Our results are co nsistent with a formal thermodynamic model for axonal growth in which removing a compressive load on axonal microtubules promotes their asse mbly, thus promoting axonal elongation.