Choline acetyltransferase-immunoreactive patches overlap specific efferent cell groups in the cat superior colliculus

Fibers containing acetylcholine (ACh) form distinct patches in the dorsal intermediate gray layer (IGL) of the cat superior colliculus (SC). Although these patches are known to overlap several afferent projections to SC, it is not known whether they are associated with specific postsynaptic cell groups. We have examined the relationship of these ACh fiber patches to specific efferent cell groups by combining retrograde transport of horseradish peroxidase (HRP) with choline acetyltransferase (ChAT) immunocytochemistry. Successful HRP injections were made into the predorsal bundle (PB), the tecto-pontine-bulbar pathway (TPB) and the cuneiform region (CFR), the inferior olive (IO), the dorsolateral pontine gray nucleus (PGD), and the pedunculopontine tegmental nucleus (PPTN). The distribution of HRP-labeled neurons which project to these targets was mapped by a computer-based microscope plotter. Distinct clusters of HRP-labeled neurons in the IGL were seen after three injections into the mesencephalic reticular formation that involved the caudal TPB and cuneiform region (CFR), and after one injection into the medial accessory nucleus of IO. As many as seven clusters of labeled neurons were found in some sections through the caudal one-half of SC after the TPB/CFR injections. Each cluster consisted of 3-20 cells, all of which were small to medium in size. In sections also tested for ChAT, the cell clusters in the TPB/CFR cases were found to overlap precisely the ACh patches in the IGL. In addition, SC neurons projecting to the IO formed clusters above the ChAT patches and in the intermediate white layer (IWL) of SC. None of the other HRP injections produced any obvious cell clusters in the deep layers of SC. These results are the first to show that specific cell groups, distinguished by size and projection site, form clusters that match the patch-like innervation of cholinergic afferents to SC. This modular organization may correspond to saccade-related cells that have also been reported to be organized into clusters in the cat SC.