In adult ferrets, retinal ganglion cells (RGCs) responsive to increased (On) or decreased (Off) illumination convey information to different cellular layers of the dorsal lateral geniculate nucleus (dLGN). These dLGN sublaminae emerge during development when RGCs are found to undergo correlated spontaneous bursting activity. Using Ca2+ imaging and intracellular dye-filling techniques, we demonstrate here that in ferret neonates, morphologically identified On and Off beta RGCs have similar burst frequencies prior to the segregation of their inputs in the dLGN, but during the segregation period, they develop distinct burst frequencies. Although the bursts of On cells and Off cells occur synchronously, On cells burst only 25%-35% of the time that Off cells do. This change in the temporal bursting patterns of On and Off RGCs may underlie the segregation of their inputs on dLGN neurons.