For many years nerve growth factor was the only factor known to influence embryonic and postnatal development of sympathetic neurons. Its deprivation by antibody neutralization or gene mutation results in extensive neuron death. Recently it has been shown that these neurons also require neurotrophin-3 for survival in the late developmental period. Using neurotrophin-3 antiserum to neutralize endogenous factor in newborn rats. Our laboratory has shown that extensive numbers of neurons are lost from both pre- and paravertebral ganglia, indicating a continuing requirement for neurotrophin-3. In the present study we sought to determine whether neurons could survive in vivo in the presence of excess amounts of either nerve growth factor or neurotrophin-3 alone. Consistent with previous findings, administration of antiserum to nerve growth factor or neurotrophin-3 to newborn rats for eight days, resulted in an extensive loss of sympathetic neurons. Interestingly, administration of neurotrophin-3 together with nerve growth factor antiserum or nerve growth factor with neurotrophin-3 antiserum reversed this neuronal loss. However the latter combination was less effective than the former. Furthermore, the ability of exogenous nerve growth factor to increase both the number and size of sympathetic neurons was prevented by the simultaneous deprivation of endogenous neurotrophin-3. Unlike nerve growth factor, exogenous neurotrophin-3 failed to rescue the naturally occurring neuronal death in these newborn rats. Further evidence for a physiological role for both nerve growth factor and neurotrophin-3 was found by the detection of both trkA and trkC immunoreactivity in neurons of the superior cervical ganglion. Taken together, these results suggest that sympathetic neurons do not have an absolute requirement for either nerve growth factor or neurotrophin-3 and that the endogenous supply of either factor alone is insufficient to support neuronal survival postnatally. However, while each factor may play similar roles in the regulation of postmitotic neuronal function, some evidence for distinct functions has been identified.