The connection between the dorsal lateral geniculate nucleus (dLGN) and area 17 of the cat is a classical model for studying thalamocortical relations. We investigated the proportion of asymmetric synapses in layer 4 of area 17 of cats formed by axons of the dLGN, because this is an important morphological parameter in understanding the impact of dLGN axons on their target neurons. Although the present consensus is that this proportion is small, the exact percentage remains in doubt. Most previous work estimated that the thalamus contributes less than 10% of excitatory synapses in layer 4, but one estimate was as high as 28%. Two issues contribute to these widely different estimates, one being the tracers used, the other being the use of biased stereological approaches. We have addressed both of these issues. Thalamic axons were labeled in vivo by injections of biotinylated dextran amine into the A lamina of the dLGN of anesthetized cats. After processing, the brain was cut serially and prepared for light and electron microscopy. The density of asymmetric synapses in the neuropil and the density of synapses formed by labeled dLGN boutons were measured by using an unbiased sampling method called the physical disector. Our counts indicate that, in the fixed cat brain, there are 5.9 × 108 ± 0.9 × 108 asymmetric synapses per cubic millimeter of layer 4 in area 17, and the dLGN input provides only 6% of all asymmetric synapses in layer 4. The vast majority of synapses of layer 4 probably originate from other neurons in area 17. J. Comp. Neurol. 516:264-276, 2009. © 2009 Wiley-Liss, Inc.