Heterogeneous output pathways link the anterior pretectal nucleus with the zona incerta and the thalamus in rat.

The anterior pretectal nucleus (APT) and the zona incerta (ZI) are diencephalic nuclei that exert a strong inhibitory influence selectively in higher order thalamic relays. The APT is also known to project to the ZI as well as the thalamus, but anatomical details of the APT-ZI projection have not been described. In the present study, the efferent pathways of the APT were examined in the APT-ZI-thalamus network by using anterograde and retrograde tracing in combination with pre- and postembedding immunocytochemical stainings and in situ hybridization. The vast majority of APT fibers selectively innervated the parvalbumin-positive, ventral part of the ZI, which contains ZI neurons with axons projecting to higher order thalamic nuclei. The APT-ZI pathway consisted of both gamma-aminobutyric acid (GABA)-negative and GABA-positive components; 38.2% of the terminals in the ZI contained GABA, and 8.6% of the projecting somata in the APT were glutamic acid decarboxylase 67 (GAD67) mRNA positive. The combination of parvalbumin immunostaining with retrograde tracing showed that strongly and weakly parvalbumin-positive as well as parvalbumin-negative neurons were all among the population of APT cells projecting to the ZI. Similar heterogeneity was found among the APT cells projecting to the thalamus. Double retrograde tracing from higher order thalamic nuclei and their topographically matched ZI regions revealed hardly any APT neuron with dual projections. Our data suggest that both ZI and the higher order thalamic relays are innervated by distinct, physiologically heterogeneous APT neurons. These various efferent pathways probably interact via the rich recurrent collaterals of the projecting APT cells. Therefore, the powerful, GABAergic APT and ZI outputs to the thalamus are apparently co-modulated in a synergistic manner via dual excitatory and inhibitory APT-ZI connections. Copyright 2007 Wiley-Liss, Inc.