Retinal bipolar cells are the first ‘projection neurons’ of the vertebrate visual system—all of the information needed for vision is relayed by this intraretinal connection. Each of the at least 13 distinct types of bipolar cells systematically transforms the photoreceptor input in a different way, thereby generating specific channels that encode stimulus properties, such as polarity, contrast, temporal profile and chromatic composition. As a result, bipolar cell output signals represent elementary ‘building blocks’ from which the microcircuits of the inner retina derive a feature-oriented description of the visual world.

SUMMARY The retina extracts visual features for transmission to the brain. Different types of bipolar cell split the photoreceptor input into parallel channels and provide the excitatory drive for downstream visual circuits. Anatomically and genetically, mouse bipolar cell types have been described at great detail, but a similarly deep understanding of their functional diversity is lacking. By imaging light-driven glutamate release from more than 13,000 bipolar cell axon terminals in the intact retina, we here show that bipolar cell functional diversity is generated by the interplay of dendritic excitatory inputs and axonal inhibitory inputs. The resultant centre and surround components of bipolar cell receptive fields interact to decorrelate bipolar cell output in the spatial and temporal domain. Our findings highlight the importance of inhibitory circuits in generating functionally diverse excitatory pathways and suggest that decorrelation of parallel visual pathways begins already at the second synapse of the mouse visual system.

For efficient coding, sensory systems need to adapt to the distribution of signals to which they are exposed. In vision, natural scenes above and below the horizon differ in the distribution of chromatic and achromatic features. Consequently, many species differentially sample light in the sky and on the ground using an asymmetric retinal arrangement of short- (S, "blue") and medium- (M, "green") wavelength-sensitive photoreceptor types. Here, we show that in mice this photoreceptor arrangement provides for near-optimal sampling of natural achromatic contrasts. Two-photon population imaging of light-driven calcium signals in the synaptic terminals of cone-photoreceptors expressing a calcium biosensor revealed that S, but not M cones, preferred dark over bright stimuli, in agreement with the predominance of dark contrasts in the sky but not on the ground. Therefore, the different cone types do not only form the basis of "color vision," but in addition represent distinct (achromatic) contrast-selective channels.

In the mouse retina, three different types of photoreceptors provide input to 14 bipolar cell (BC) types. Classically, most BC types are thought to contact all cones within their dendritic field; ON-BCs would contact cones exclusively via so-called invaginating synapses, while OFF-BCs would form basal synapses. By mining publically available electron microscopy data, we discovered interesting violations of these rules of outer retinal connectivity: ON-BC type X contacted only ~20% of the cones in its dendritic field and made mostly atypical non-invaginating contacts. Types 5T, 5O and 8 also contacted fewer cones than expected. In addition, we found that rod BCs received input from cones, providing anatomical evidence that rod and cone pathways are interconnected in both directions. This suggests that the organization of the outer plexiform layer is more complex than classically thought.DOI: http://dx.doi.org/10.7554/eLife.20041.001

Horizontal cells are interneurons of the vertebrate retina that exhibit strong electrical and tracer coupling but the identity of the channel-forming connexins has remained elusive. Here we show that horizontal cells of the mouse retina express connexin57 (Cx57). We have generated Cx57-deficient mice by replacing the Cx57 coding region with a lacZ reporter gene, expressed under control of the endogenous Cx57 promoter. These mice were fertile and showed no obvious anatomical or behavioural abnormalities. Cx57 mRNA was expressed in the retina of wild-type littermates but was absent from the retina of Cx57-deficient mice. Previously reported results that the Cx57 gene was very weakly expressed in several other mouse tissues turned out to be unspecific. Cx57 mRNA is abundantly expressed in the retina and weakly in the thymus of adult mice but absent in all other adult tissues tested, including brain. Furthermore, Cx57 is expressed in embryonic kidney at E16.5 to E18.5 days post-conception, as indicated by the pattern of lacZ expression. Within the retina, lacZ signals were assigned exclusively to horizontal cells based on co-localization with cell-type-specific marker proteins. Microinjection of Neurobiotin into horizontal cells of isolated retinae revealed less than 1% of tracer coupling in Cx57-deficient retinae compared with wild-type controls. Cx57 is the first connexin identified in mammalian horizontal cells and the first connexin whose expression is apparently restricted to only one type of neuron.

Gap junctions in AII amacrine cells of mammalian retina participate in the coordination of the rod and cone signaling pathway involved in visual adaptation. Upon stimulation by light, released dopamine binds to D 1 receptors on AII amacrine cells leading to increased intracellular cAMP (cyclic adenosine monophosphate) levels. AII amacrine cells express the gap junctional protein connexin36 (Cx36). Phosphorylation of Cx36 has been hypothesized to regulate gap junctional activity of AII amacrine cells. However, until now in vivo phosphorylation of Cx36 has not been reported. Indeed, it had been concluded that Cx36 in bovine retina is not phosphorylated, but in vitro phosphorylation for Cx35, the bass ortholog of Cx36, had been shown. To clarify this experimental discrepancy, we examined protein kinase A (PKA)-induced phosphorylation of Cx36 in mouse retina as a possible mechanism to modulate the extent of gap junctional coupling. The cytoplasmic domains of Cx36 and the total Cx36 protein were phosphorylated in vitro by PKA. Mass spectroscopy revealed that all four possible PKA consensus motifs were phosphorylated; however, domains point mutated at the sites in question showed a prevalent usage of Ser-110 and Ser-293. Additionally, we demonstrated that Cx36 was phosphorylated in cultured mouse retina. Furthermore, activation of PKA increased the level of phosphorylation of Cx36. cAMP-stimulated, PKA-mediated phosphorylation of Cx36 protein was accompanied by a decrease of tracer coupling between AII amacrine cells. Our results link increased phosphorylation of Cx36 to down-regulation of permeability through gap junction channels mediating light adaptation in the retina.The mammalian retina is a structure with three neuronal layers and two synaptic layers in which retinal neurons are not only forming numerous chemical synaptic contacts but also electrically and chemically coupled networks via gap junction channels. Gap junctions were reported in rod and cone photoreceptor cells, cone bipolar cells, horizontal cells, various subtypes of amacrine cells, and ganglion cells (1). Interneuronal communication through gap junction channels is essential for the rod pathway under conditions of scotopic illumination. Visual information from the rods is carried via rod bipolar cells to AII amacrine cells that form heterologous electrical synapses with ON cone bipolar cells, thus transferring rod signals to ganglion cells (2-4). Junctional hemichannels on the AII amacrine cell side consist of Cx36, 2 whereas Cx36 and/or Cx45 protein form electrical synapses on the bipolar cell side (5-7). Additionally, AII amacrine cells form homotypic gap junctions to neighboring AII amacrine cells, involving Cx36 (2, 3, 8 -10). Under scotopic conditions, the AII network only shows weak coupling and thus supports optimal transfer of the rod signal to the ON cone bipolar cells. Under mesopic light conditions, AII amacrine cells form an extensively coupled network. Pooling visual signals under this light condition increases sensitivity of AII ama...

Background: Neurons receive excitatory synaptic inputs that are distributed across their dendritic arbors at densities and with spatial patterns that influence their output. How specific synaptic distributions are attained during development is not well understood. The distribution of glutamatergic inputs across the dendritic arbors of mammalian retinal ganglion cells (RGCs) has long been correlated to the spatial receptive field profiles of these neurons. Thus, determining how glutamatergic inputs are patterned onto RGC dendritic arbors during development could provide insight into the cellular mechanisms that shape their functional receptive fields.

Sensory neurons with common function are often non-randomly arranged and form dendritic territories that exhibit little overlap or tiling. Repulsive homotypic interactions underlie such patterns in cell organization in invertebrate neurons. In mammalian retinal horizontal cells, however, it is unclear how dendro-dendritic repulsive interactions can produce a non-random distribution of cells and their spatial territories because mature horizontal cell dendrites overlap substantially. By imaging developing mouse horizontal cells, we found that upon reaching their final laminar positions, these cells transiently elaborate vertical neurites that form non-overlapping columnar territories. Targeted cell ablation revealed that the vertical neurites engage in homotypic interactions resulting in tiling of neighboring cells prior to establishment of their dendritic fields. This developmental tiling of transient neurites correlates with the emergence of a non-random distribution of the cells, and could represent a mechanism that organizes neighbor relationships and territories of neurons of the same type before circuit assembly.