Expression of Ca2+-Binding Buffer Proteins in the Human and Mouse Retinal Neurons.

Ca2+-binding buffer proteins (CaBPs) are widely expressed by various neurons throughout the central nervous system (CNS), including the retina. While the expression of CaBPs by photoreceptors, retinal interneurons and the output ganglion cells in the mammalian retina has been extensively studied, a general description is still missing due to the differences between species, developmental expression patterns and study-to-study discrepancies. Furthermore, CaBPs are occasionally located in a compartment-specific manner and two or more CaBPs can be expressed by the same neuron, thereby sharing the labor of Ca2+ buffering in the intracellular milieu. This article reviews this topic by providing a framework on CaBP functional expression by neurons of the mammalian retina with an emphasis on human and mouse retinas and the three most abundant and extensively studied buffer proteins: parvalbumin, calretinin and calbindin.

The agonistic TSPO ligand XBD173 attenuates the glial response thereby protecting inner retinal neurons in a murine model of retinal ischemia.

BACKGROUND: Ligand-driven modulation of the mitochondrial translocator protein 18 kDa (TSPO) was recently described to dampen the neuroinflammatory response of microglia in a retinal light damage model resulting in protective effects on photoreceptors. We characterized the effects of the TSPO ligand XBD173 in the postischemic retina focusing on changes in the response pattern of the major glial cell types of the retina-microglia and Müller cells. METHODS: Retinal ischemia was induced by increasing the intraocular pressure for 60 min followed by reperfusion of the tissue in mice. On retinal cell types enriched via immunomagnetic separation expression analysis of TSPO, its ligand diazepam-binding inhibitor (DBI) and markers of glial activation were performed at transcript and protein level using RNA sequencing, qRT-PCR, lipid chromatography-mass spectrometry, and immunofluorescent labeling. Data on cell morphology and numbers were assessed in retinal slice and flatmount preparations. The retinal functional integrity was determined by electroretinogram recordings. RESULTS: We demonstrate that TSPO is expressed by Müller cells, microglia, vascular cells, retinal pigment epithelium (RPE) of the healthy and postischemic retina, but only at low levels in retinal neurons. While an alleviated neurodegeneration upon XBD173 treatment was found in postischemic retinae as compared to vehicle controls, this neuroprotective effect of XBD173 is mediated putatively by its action on retinal glia. After transient ischemia, TSPO as a marker of activation was upregulated to similar levels in microglia as compared to their counterparts in healthy retinae irrespective of the treatment regimen. However, less microglia were found in XBD173-treated postischemic retinae at 3 days post-surgery (dps) which displayed a more ramified morphology than in retinae of vehicle-treated mice indicating a dampened microglia activation. Müller cells, the major retinal macroglia, show upregulation of the typical gliosis marker GFAP. Importantly, glutamine synthetase was more stably expressed in Müller glia of XBD173-treated postischemic retinae and homeostatic functions such as cellular volume regulation typically diminished in gliotic Müller cells remained functional. CONCLUSIONS: In sum, our data imply that beneficial effects of XBD173 treatment on the postischemic survival of inner retinal neurons were primarily mediated by stabilizing neurosupportive functions of glial cells.

Short-wavelength cone-opponent retinal ganglion cells in mammals.

In all of the mammalian species studied to date, the short-wavelength-sensitive (S) cones and the S-cone bipolar cells that receive their input are very similar, but the retinal ganglion cells that receive synapses from the S-cone bipolar cells appear to be quite different. Here, we review the literature on mammalian retinal ganglion cells that respond selectively to stimulation of S-cones and respond with opposite polarity to longer wavelength stimuli. There are at least three basic mechanisms to generate these color-opponent responses, including: (1) opponency is generated in the outer plexiform layer by horizontal cells and is conveyed to the ganglion cells via S-cone bipolar cells, (2) inputs from bipolar cells with different cone inputs and opposite response polarity converge directly on the ganglion cells, and (3) inputs from S-cone bipolar cells are inverted by S-cone amacrine cells. These are not mutually exclusive; some mammalian ganglion cells that respond selectively to S-cone stimulation seem to utilize at least two of them. Based on these findings, we suggest that the small bistratified ganglion cells described in primates are not the ancestral type, as proposed previously. Instead, the known types of ganglion cells in this pathway evolved from monostratified ancestral types and became bistratified in some mammalian lineages.

Metabolic profiling of the mouse retina using amino acid signatures: insight into developmental cell dispersion patterns.

Pattern recognition has been used for the complete and statistically rigid classification of retinal neurons in vertebrates such as the adult cat, primate, rat and goldfish. Here, we label the mouse retina with antibodies against seven amino acids and use pattern recognition to characterize distinct retinal neurochemical cell classes based on their unique amino acid signatures. We followed the development of the cell classes in the X-inactivation transgenic mouse expressing the lacZ reporter gene on one X-chromosome. This mouse allows clonally related cells to be identified through differential ß-galactosidase activity due to random X-chromosome inactivation. Pattern recognition analysis partitioned the retina into nine neuronal classes at birth, increasing to 19 classes at eye opening and 26 classes by adulthood. Emergence of new cell classes was partly attributed to new neuron types and partly to the splitting of classes from early ages from refinement of their amino acid profiles. All six GABAergic amacrine cell classes and most ganglion cell classes appeared by P7 whilst all the glycinergic amacrine cell classes did not appear till adulthood. Separable bipolar cell classes were not detected till eye opening. Photoreceptor cell classes were detected at P3 but inner and outer segments did not form separable classes until adulthood. More importantly, we show that cells which share common amino acid profiles also shared cell dispersion patterns. GABAergic amacrine cell classes with conventional and displaced counterparts transgressed clonal boundaries whereas GABAergic amacrine cell classes found exclusively in the inner nuclear layer and all glycinergic amacrine cell classes did not transgress. Ganglion cells displayed both dispersion patterns. This study provides a comprehensive neurochemical atlas of the developing mouse retina, tracking the amino acid levels within distinct neuronal populations and highlighting unique migratory patterns within subpopulations of inner retinal neurons.

Light adaptation alters the source of inhibition to the mouse retinal OFF pathway.

Sensory systems must avoid saturation to encode a wide range of stimulus intensities. One way the retina accomplishes this is by using both dim-light-sensing rod and bright-light-sensing cone photoreceptor circuits. OFF cone bipolar cells are a key point in this process, as they receive both excitatory input from cones and inhibitory input from AII amacrine cells via the rod pathway. However, in addition to AII amacrine cell input, other inhibitory inputs from cone pathways also modulate OFF cone bipolar cell light signals. It is unknown how these inhibitory inputs to OFF cone bipolar cells change when switching between rod and cone pathways or whether all OFF cone bipolar cells receive rod pathway input. We found that one group of OFF cone bipolar cells (types 1, 2, and 4) receive rod-mediated inhibitory inputs that likely come from the rod-AII amacrine cell pathway, while another group of OFF cone bipolar cells (type 3) do not. In both cases, dark-adapted rod-dominant light responses showed a significant contribution of glycinergic inhibition, which decreased with light adaptation and was, surprisingly, compensated by an increase in GABAergic inhibition. As GABAergic input has distinct timing and spatial spread from glycinergic input, a shift from glycinergic to GABAergic inhibition could significantly alter OFF cone bipolar cell signaling to downstream OFF ganglion cells. Larger GABAergic input could reflect an adjustment of OFF bipolar cell spatial inhibition, which may be one mechanism that contributes to retinal spatial sensitivity in the light.

Role of neurotransmitter receptors in mediating light-evoked responses in retinal interplexiform cells.

Interplexiform (IP) cells are a long-neglected group of retinal neurons the function of which is yet to be determined. Anatomical study indicates that IP cells are located in the inner nuclear layer, juxtaposed with the third-order neurons. However, the synaptic transmission of IP cells in the inner retina is poorly understood. Using whole cell patch-clamp and pharmacological techniques, we extensively studied synaptic receptors in IP cells. The IP cells in amphibian retinal slices were identified by electrical and morphological properties with voltage-clamp recording and Lucifer yellow dialysis. We find that light-evoked excitatory postsynaptic currents (L-EPSCs) are mediated by AMPA and N-methyl-d-aspartate receptors in IP cells. Although both receptors contributed to the amplitude and kinetics of L-EPSCs, AMPA receptor desensitization substantially shaped L-EPSCs in the neurons, similar to those found in the third-order neurons. The light-evoked inhibitory postsynaptic currents (L-IPSCs) in IP cells were primarily mediated by strychnine-sensitive glycine receptors with a small component of GABA(C) receptors. GABA(C) receptor rho2 subunits were detected in IP cells with single-cell RT-PCR assays. Expression of GABA(C) receptors is one of the special features for IP cells, distinct from most of the third-order neurons that depend on GABA(A) and glycine receptors to relay the inhibitory signals. However, GABA(A) receptors in IP cells acted like nonsynaptic receptors that were activated by exogenous GABA application. Furthermore, L-IPSCs in IP cells were inhibited by the serial inhibitions between amacrine cells in the inner retina. In addition, application of neurotransmitters on the axon terminals of IP cells had no significant current generated in the cells, indicating that the synaptic inputs of IP cells are mainly from the inner retina. This study demonstrates the important role that light signals are encoded by both experiment of inhibitory receptors in IP cells.