A double-antibody heterologous radioimmunoassay (RIA) was developed to measure plasma LH values in hedgehogs. This RIA system used anti-rat LH serum and rabbit LH (AFP-559B) for radioiodination and as standard. The accuracy of the method was evaluated and indicated the ability to detect various relative concentrations of LH in plasma. The minimum detectable dose was 0.2 ng/ml. The intra- and inter-assay coefficients of variation were 4.2 and 7.9% respectively. Biological tests, e.g. effect of castration, effect of castration + testosterone implant and GnRH administration, confirmed that this method was suitable to determine subsequent changes in pituitary gonadotrophic activity in the hedgehog. LH concentrations were determined in blood samples obtained during 1 year: (a) each month, at 4-h intervals during 24 h, from different groups of unanaesthetized animals fitted with a catheter and (b) twice a month, under a light anaesthesia, from the same group of 6 animals. During the year: (1) the range of LH change was narrow (minimum values congruent to 0.25 ng/ml and maximum values congruent to 2.00 ng/ml); (2) the 24-h LH patterns did not exhibit any daily rhythm; (3) a clear annual rhythm was observed with the highest values from February to April and the lowest values in October and November. LH decreased rapidly at the end of summer and increased progressively from December to February, during hibernation. In these experiments, it was not possible to determine the characteristics of LH release patterns in the hedgehog but individual profiles indicated clearly the episodic secretion of LH, particularly during the highest pituitary activity period. During the year, a close relationship between the seasonal cycles of plasma LH and testosterone was observed.