Factors involved in the stimulation of parasympathetic nerve outgrowth.

Nerve growth from the mouse parasympathetic submandibular ganglion is stimulated by the developing target epithelium. To investigate the nature of this trophic influence, homogenates of salivary glands, gland-conditioned medium, and formalin-fixed glands were assayed for ability to elicit parasympathetic axon extension in tissue culture. Neither homogenates nor conditioned medium stimulated axon outgrowth from submandibular ganglia. However, when ganglia were added to glands in which protein synthesis and cell movement had been halted by formalin fixation, stimulation of outgrowth into the tissue was observed. Stimulation of axonal growth occurred after hyaluronidase and collagenase treatment of the glands, but not after treatment with proteases or with heat. Moreover, prolonged formalin fixation destroyed the glandular ability to elicit axon elongation. Intact ganglia cultured with whole live submandibular glands in the presence of low levels of hyaluronidase or collagenase showed extensive axon outgrowth despite disruption of the normal morphogenetic pattern of both epithelium and axons. Our results suggest that stimulation of axon outgrowth does not directly depend on the concomitant metabolic or morphogenetic activity of the epithelium, but is caused by some epithelial product, probably a protein.