Acute insulin administration causes a disparity between the onset of estrous behavior and the LH surge in ovary-intact ewes. To examine the considerable variation in responses, in the present study we used a large number of animals to confirm findings with insulin, and examine whether endotoxin has the same effect. During the breeding season, follicular phases of intact ewes were synchronized with progesterone vaginal pessaries and received saline vehicle (n=22; controls), insulin (4 IU/kg; n=21 ewes) or endotoxin (LPS; 100 ng/kg; n=10) at 28 h after progesterone withdrawal (time zero). In controls, the LH surge onset occurred at 36.5 +/- 5.7 h and were first mounted by a ram at 38.2 +/- 1.8 h, but there was a delay of 17.6 h (P<0.001) and 7.2 h (P<0.05), respectively, in half the insulin-treated animals ('insulin-delayed') but not in the other half; and a delay of 22.5 h (P<0.001) and 20.7 h (P<0.001), respectively, in all LPS-treated animals. Plasma estradiol concentrations decreased after both stressors, and remained low for a period of time equivalent to the LH surge delay (P<0.001; Rs-q=78%). Cortisol increased for 12 h after treatment in both insulin subgroups and the LPS group (P<0.05); whereas progesterone increased in the insulin-delayed and LPS groups from 4.0 +/- 0.5 ng/ml and 5.3 +/- 1.0 ng/ml to a maximum of 5.7 +/- 0.3 ng/ml and 8.8 +/- 1.6 ng/ml, respectively (P<0.05 for both comparisons). Plasma triglycerides were measured to assess insulin resistance, but concentrations were similar before and after treatment (0.25 +/- 0.01 mmol/l versus 0.21 +/- 0.01 and 0.25 +/- 0.01 mmol/l versus 0.26 +/- 0.01 mmol/l in the insulin-non delayed and insulin delayed subgroups, respectively). Therefore, we hypothesize that a) when an acute stressor is applied during the late follicular phase, the duration of the LH surge delay is related to the duration of estradiol signal disruption b) cortisol is not the key disruptor of the LH surge after insulin, c) insulin (but not LPS) can separate the onsets of LH surge and estrus by approximately 10 h, providing a model to identify the specific neuronal systems that control behavior distinct from those initiating the GnRH surge. (C) 2011 Elsevier Inc. All rights reserved.