Whether or not adrenal medullary (chromaffin) cells which respond to nerve growth factor (NGF) both in vitro and in vivo require NGF for their normal development is controversial. Systemic deprivation of endogenous NGF by injection of anti-NGF antibodies into rat fetuses or by transfer of anti-NGF to the offspring of autoimmunized mothers has provided conflicting results. We have reinvestigated the effects of a specific antiserum to NGF on the morphology, catecholamine (CA) and neuropeptide (Met-enkephalin, Met-ENK; substance P, SP) content, and choline acetyltransferase (ChAT) activity of the rat adrenal medulla. Fetuses were injected with anti-NGF antibodies on day 17 of gestation and postnatally at daily intervals for 7 days. The histological appearance of adrenal medullae of anti-NGF injected animals was not altered as compared to controls. Ultrastructurally, no degenerative changes or developmental retardation of chromaffin cells could be detected. However, numbers of chromaffin granules per micron 2 of cytoplasmic area were greater and the mean diameters of the cores of adrenaline storage granules were smaller in antibody-treated than in control animals. CA and SP content, ratios of adrenaline to noradrenaline and ChAT activities were identical in anti-NGF-treated and control animals. Anti-NGF antibodies caused a reduction of adrenal Met-ENK by 40% as compared to controls. Superior cervical ganglia from the same animals were used to document immunosympathectomy induced by the antiserum. They displayed the well-established structural alterations and a marked reduction of the CA content. We conclude that administration of anti-NGF antibodies to embryonic and early postnatal rats induces only subtle changes in the ultramorphology of chromaffin cells without altering the development of normal CA levels. The small, yet significant effects of anti-NGF antibodies on adrenal Met-ENK, however, may suggest a role for endogenous NGF in the regulation of opioid peptide metabolism in developing chromaffin cells.