The retina contains ganglion cells (RGCs) that respond selectively to objects moving in particular directions. Individual members of a group of ON-OFF direction-selective RGCs (ooDSGCs) detect stimuli moving in one of four directions: ventral, dorsal, nasal or temporal. Despite this physiological diversity, little is known about subtype-specific differences in structure, molecular identity and projections. To seek such differences, we characterized mouse transgenic lines that selectively mark ooDSGCs preferring ventral or nasal motion as well as a line that marks both ventral- and dorsal-preferring subsets. We then used the lines to identify cell surface molecules, including Cadherin 6, Collagen25a1, and Matrix metalloprotease 17, that are selectively expressed by distinct subsets of ooDSGCs. We also identify a neuropeptide, CART, that distinguishes all ooDSGCs from other RGCs. Together, this panel of endogenous and transgenic markers distinguishes the four ooDSGC subsets. Patterns of molecular diversification occur before eye-opening and are therefore experience-independent. They may help explain how the four subsets obtain distinct inputs. We also demonstrate differences among subsets in their dendritic patterns within the retina and their axonal projections to the brain. Differences in projections indicate that information about motion in different directions is sent to different destinations.

SUMMARY How are sensory representations in the brain influenced by the state of an animal? Here we use chronic two-photon calcium imaging to explore how wakefulness and experience shape odor representations in the mouse olfactory bulb. Comparing the awake and anesthetized state, we show that wakefulness greatly enhances the activity of inhibitory granule cells and makes principal mitral cell odor responses more sparse and temporally dynamic. In awake mice, brief repeated odor experience leads to a gradual and long-lasting (months) weakening of mitral cell odor representations. This mitral cell plasticity is odor-specific, recovers gradually over months and can be repeated with different odors. Furthermore, the expression of this experience-dependent plasticity is prevented by anesthesia. Together, our results demonstrate the dynamic nature of mitral cell odor representations in awake animals, which is constantly shaped by recent odor experience.

In many parts of the nervous system, neuronal somata display orderly spatial arrangements1. In the retina, neurons of numerous individual subtypes form regular arrays called mosaics: they are less likely to be near neighbors of the same subtype than would occur by chance, resulting in “exclusion zones” that separate them1-4. Mosaic arrangements provide a mechanism to distribute each cell type evenly across the retina, ensuring that all parts of the visual field have access to a full set of processing elements2. Remarkably, mosaics are independent of each other: while a neuron of one subtype is unlikely to be adjacent to another of the same subtype, there is no restriction on its spatial relationship to neighboring neurons of other subtypes5. This independence has led to the hypothesis that molecular cues expressed by specific subtypes pattern mosaics by mediating homotypic (within-subtype) short-range repulsive interactions1,4-9. To date, however, no molecules have been identified that show such activity, so this hypothesis remains untested. Here, we demonstrate that two related transmembrane proteins, MEGF10 and MEGF11, play critical roles in formation of mosaics by two retinal interneuron subtypes, starburst amacrine cells (SACs) and horizontal cells (HCs). MEGF10/11 and their invertebrate relatives C. elegans CED-1 and Drosophila Draper, have hitherto been studied primarily as receptors necessary for engulfment of debris following apoptosis or axonal injury10-14. Our results demonstrate that members of this gene family can also serve as subtype-specific ligands that pattern neuronal arrays.

Summary For reliable stimulus identification, sensory codes have to be robust by including redundancy to combat noise, but redundancy sacrifices coding efficiency. To address how experience affects the balance between the robustness and efficiency of sensory codes, we probed odor representations in the mouse olfactory bulb during learning over a week, using longitudinal two-photon calcium imaging. When mice learned to discriminate between two dissimilar odorants, responses of mitral cell ensembles to the two odorants gradually became less discrete, increasing the efficiency. In contrast, when mice learned to discriminate between two very similar odorants, the initially overlapping representations of the two odorants became progressively decorrelated, enhancing the robustness. Qualitatively similar changes were observed when the same odorants were experienced passively, a condition that would induce implicit perceptual learning. These results suggest that experience adjusts odor representations to balance the robustness and efficiency depending on the similarity of the experienced odorants.

Most regions of the central nervous system contain numerous subtypes of inhibitory interneurons that play specialized roles in circuit function. In mammalian retina, the ~30 subtypes of inhibitory interneurons called amacrine cells (ACs) are generally divided into two groups: wide/medium-field GABAergic and narrow-field glycinergic, which mediate lateral and vertical interactions, respectively, within the inner plexiform layer. We used expression profiling and mouse transgenic lines to identify and characterize two closely-related narrow-field AC subtypes. Both arise postnatally and one, surprisingly, is neither glycinergic nor GABAergic (nGnG). Two transcription factors selectively expressed by these subtypes, Neurod6 and Satb2, regulate a postmitotic cell fate choice between them. Satb2 induces Neurod6, which persists in nGnG ACs and promotes their fate, but is down-regulated in the related glycinergic AC subtype. Our results support the view that cell fate decisions made in progenitors and their progeny act together to diversify ACs.