The effects of two fully characterized, structurally distinct gonadotropins, GtH I and GtH II, on the uptake of vitellogenin (VTG) into oocytes of the rainbow trout, Oncorhynchus mykiss, were investigated both in vivo and in vitro. GtH I, injected into maturing vitellogenic females at a dose of 10 micrograms.kg body wt-1, increased the rate of [3H]VTG uptake into oocytes by more than two-fold, effectively doubling their rate of growth. Ovaries from females similarly treated with GtH II sequestered VTG at rates similar to controls. In vitro, GtH I stimulated VTG uptake in a dose-dependent manner. At a GtH I concentration of 100 ng.ml-1 and above, the rate of VTG uptake was significantly greater than that of the controls and at 1000 ng.ml-1 the rate of uptake was more than doubled. GtH II did not significantly increase VTG sequestration into isolated oocytes at concentrations up to, and including, 1000 ng.ml-1. These data provide the first evidence that GtH I has a primary function in stimulating VTG uptake and strongly support the contention that at least two functionally distinct GtHs occur in fish.