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Cultures were grown in serum\u2010free chemically defined medium and were treated with either Met\u2010enkephalin antiserum (anti\u2010Met) to immunoneutralize enkephalins, or with naloxone, a universal opioid receptor antagonist, to block receptor\u2010mediated actions of released endogenous opioids. The enzyme activities of choline acetyl\u2010transferase (ChAT) and tyrosine hydroxylase (TH) were used as markers for cholinergic and catecholaminergic phenotypic expression, respectively. We found that cultures treated with anti\u2010Met or naloxone exhibited strikingly different neuronal growth patterns as compared to controls. In addition, ChAT activity was enhanced by anti\u2010Met, and TH activity by both anti\u2010Met and naloxone. 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