Thyrotrophin-releasing hormone (TRH) was demonstrated immunocytochemically in the infundibulum of the chick embryo as early as Day 4.5 of incubation. From Days 4.5 through 19.5 of embryonic development there is a gradual increase within the developing hypothalamus in the number of TRH-positive perikarya as well as the amount of immunoreactive-TRH (IR-TRH) per cell. There are no abrupt changes in either parameter during the critical time period (Days 10.5-13.5 of incubation) in the maturation of the pituitary-thyroid axis. Thus, although TRH is probably not directly responsible for the dramatic increase in the number of thyrotrophin-producing cells which occurs in the pars distalis of 10.5- to 11.5-day-old embryos (R. C. Thommes, J. B. Martens, W. E. Hopkins, J. Caliendo, M. J. Sorrentino, and J. E. Woods (1983). Gen. Comp. Endocrinol. 51, 434-443) the marked change in the activity of the pituitary-thyroid unit at this time may well reflect the response of these newly differentiated thyrothrophs to low levels of plasma TRH. This hypothesis is supported by the observations that between Days 10.5 and 11.5 the hypothalamic-adenohypophyseal-thyroid (HAT) axis is first responsive to cold (R. C. Thommes, J. B. Martens, J. B. Hopkins, D. A. Griesbach, D. J. Williams, M. J. Sorrentino, P. Wernke, and J. E. Woods. In "Proceedings, Ninth International Symposium on Comparative Endocrinology Hong Kong, 7-11 December 1981" (B. Lofts, ed.). Hong Kong Univ. Press, Hong Kong, in press) and also that the pituitary-thyroid unit exhibits a marked increase in its sensitivity to exogenous TRH (R. C. Thommes, D. J. Williams, and J. E. Woods (1984). Gen. Comp. Endocrinol. 55, 275-279).