Neurotransmitter secretion from the nerve terminal is mediated by the fusion of synaptic vesicles with the plasma membrane. It is generally believed that neurotransmitter release in mature synapses is localized to the presynaptic nerve terminals. To probe the topology of neurotransmitter secretion along developing axons in culture, we recorded membrane currents from myocytes manipulated into contact with axons. At the early stages of growth, exocytotic events were detected along the axon as well as at the growth cone. At the later stages of growth, neurotransmitter secretion adopted the form of a smooth proximodistal gradient, with the highest level of activity at the growth cone region. Our results reveal the existence of a previously undetected early stage of axonal growth and suggest developmental regulation in the pattern of neurotransmitter secretion along the growing nerve processes.