The effect of manual teat stimulation (milking paradigm) on release of oxytocin, epinephrine and norepinephrine was studied in (1) 15 heifers at 100, 150, 200 and 250 d of gestation and at 30 and 90 d of lactation (during machine milking) and (2) simultaneously in six heifers at less than 100 d and in six heifers at greater than 200 d of gestation. Oxytocin responses to teat stimulation, including peak heights and area under the response curves, at 150, 200 and 250 d or at greater than 200 d of gestation were similar and were significantly greater than responses at 100 d or at less than 100 d. Responses to milking were lower at 90 d compared to responses at 30 d. Catecholamines were measured only during gestation and were generally not affected by teat stimulation. Epinephrine levels were significantly higher at 200 and 250 d compared to levels at 100 and 150 d. Baseline oxytocin concentrations and responses to teat stimulation were greatest at 150 d of gestation when epinephrine levels were still low, suggesting that stimulatory mechanisms responsible for the release of oxytocin develop and/or are expressed prior to the development of inhibitory sympathetic mechanisms. For norepinephrine, linear analyses did not show significant responses to teat stimulation overall. However, elevated norepinephrine responses (greater than .2 pmol/ml) following teat stimulation were seen in 28 of 51 trials, and large oxytocin responses (greater than 75 pg/ml/min) were seen predominantly only when norepinephrine responses were low (less than = .2 pmol/ml).