The recent development of an ultrasensitive immunofluorometric rat LH assay makes possible evaluation of pulsatile LH secretion in intact male rats under physiological conditions of minimal volume blood sampling without requiring orchidectomy. Specifically, we applied this assay to determine the effect of macronutrient restriction on pulsatile LH secretion in the presence or absence of testes. In testes-intact rats, halving of food intake for 7 days while maintaining micronutrient supply caused a reduction of mean, maximal, and basal LH levels and LH pulse amplitude (all P < 0.05) compared with those in ad libitum fed controls. The loss of body weight was positively correlated with decreases in mean LH level, pulse amplitude, and area under the curve (all P < 0.009). In contrast, the same food restriction in castrated rats caused an increase in pulse length and area under the curve and a decrease in pulse frequency, but did not change mean, maximal, and basal LH levels or LH pulse amplitude compared to castrated ad libitum fed controls (all P < 0.02). The observed positive correlations between body weight and the LH secretion parameters in intact rats were absent or reversed in castrated rats. This study demonstrates qualitatively different effects of macronutrient restriction on pulsatile LH secretion in castrated and intact rats, indicating that it is not necessary valid to extrapolate consequences of undernutrition on LH secretion from castrate to intact male rats. We conclude that undernutrition-induced inhibition of LH secretion involves both an indirect suppression of LH secretion via amplification of endogenous testicular negative feedback as well as more direct suppression of GnRH release.