The anterior forebrain pathway in songbirds is a specialization of the avian basal ganglia pathway and is prominent in males that sing, but seem to be absent or incomplete in females that do not sing. We studied the connectivity in females in the in vitro slice preparation by applying the tracer Fluoro Ruby, biotinylated dextran amine, and cholera toxin B. We identified (1) retrograde labeled neurons in the lateral magnocellular nucleus of the anterior nidopallium (LMAN) projecting to the medial striatum (MSt), and (2) we identified fibers in the MSt labeled by anterograde transport after tracer injection into LMAN. Our data clearly demonstrate the existence of a cortico-basal ganglia pathway in female birds.