OFF Bipolar Cells Research Articles

Disinhibition Combines with Excitation to Extend the Operating Range of the OFF Visual Pathway in Daylight

Cone signals divide into parallel ON and OFF bipolar cell pathways, which respond to objects brighter or darker than the background and release glutamate onto the corresponding type of ganglion cell. It is assumed that ganglion cell excitatory responses are driven by these bipolar cell synapses. Here, we report an additional mechanism: OFF ganglion cells were driven in part by the removal of synaptic inhibition (disinhibition). The disinhibition played a relatively large role in driving responses at low contrasts. The disinhibition persisted in the presence of CNQX and d-AP-5. Furthermore, the CNQX/d-AP-5-resistant response was blocked by l-AP-4, meclofenamic acid, quinine, or strychnine but not by bicuculline. Thus, the disinhibition circuit was driven by the ON pathway and required gap junctions and glycine receptors but not ionotropic glutamate or GABA(A) receptors. These properties implicate the AII amacrine cell, better known for its role in rod vision, as a critical circuit element through the following pathway: cone --> ON cone bipolar cell --> AII cell --> OFF ganglion cell. Rods could also drive this circuit through their gap junctions with cones. Thus, to light decrement, AII cells, driven by electrical synapses with ON cone bipolar cells, would hyperpolarize and reduce glycine release to excite OFF ganglion cells. To light increment, the AII circuit would directly inhibit OFF ganglion cells. These results show a new role for disinhibition in the retina and suggest a new role for the AII amacrine cell in daylight vision.

Local Neuroretinal Function during Acute Hypoxia in Healthy Older People

To measure the effect of acute, mild, systemic hypoxia on neuroretinal function in two different age groups. The slow-flash paradigm of the multifocal electroretinogram (mfERG) was measured in two different oxygenation levels. Twelve participants of two age groups (28 +/- 4 and 55 +/- 5 years) breathed room air (normoxia) or 12% oxygen (hypoxia). During normoxia (SaO(2) = 98%) there was a significantly different effect between the younger and older participants on the mfERG positive waveform components (N1P1 amplitudes and P1 implicit times; P < 0.001). Hypoxia (SaO(2) = 90%) significantly decreased N1P1 response density amplitudes in the central retinal field in both groups, with a larger effect in the older group (P < 0.001). There was no significant change in function of the cellular generators of oscillatory potentials (OPs). The results extend recent findings by showing a greater effect of hypoxia on ON- and OFF-bipolar cell function in healthy older persons than in younger persons, but no effect on OPs during moderate acute hypoxia in either age group.

Multifocal Electroretinogram in Rhodopsin P347L Transgenic Pigs

Neural ectopic rewiring in retinal degeneration such as retinitis pigmentosa (RP) may form functional synapses between cones and rod bipolar cells that cause atypical signal processing. In this study, the multifocal electroretinograms (mfERGs) of a large animal model of RP, the rhodopsin P347L transgenic (Tg) pig, were measured to examine the sources and nature of altered signal processing. mfERG responses from a 6-week-old Tg pig were recorded before and after sequential application of tetrodotoxin (TTX), N-methyl-D-aspartate (NMDA), 2-amino-4-phosphonobutyric acid (APB), and cis-2,3-piperidinedicarboylic acid (PDA), to identify contributions to the retinal signal from inner retinal neurons, the ON-pathway, the OFF-pathway, and photoreceptors. The mfERG response contributions from different retinal components of in the Tg eyes were estimated and compared with control data from eyes of age-matched wild-type (WT) pigs. There was a prominent difference in the estimates of the inner retinal response and ON-bipolar cell pathway contribution between the Tg and WT mfERG responses. In particular, the early components of the inner retinal contribution were obviously altered in the Tg mfERG. The inner retinal components at approximately 24 and 40 ms appeared to be inverted. Differences in the estimates of OFF-bipolar cell pathway contributions were minimal. There was no change in cone cell responses in the Tg mfERG. In Tg retinas, ectopic synapses formed between cones and rod bipolar cells probably altered signal processing of the ON-bipolar cell pathway. In response to the altered visual signal input from the outer retina, signal processing in inner retinal neurons was also modified.

Photopic Electroretinograms of mGluR6-Deficient Mice

Purpose: To study the properties of the photopic electroretinograms (ERGs) of the metabotropic glutamate receptor subtype 6 (mGluR6)-deficient mice and to investigate the contribution of cone ON-and OFF-pathways to the mouse photopic ERGs. Methods: Photopic ERGs were recorded from mGluR6-deficient and wild-type mice. Photopic ERGs were also recorded after an intravitreous injection ofcis-2,3 piperidine dicarboxylic acid (PDA) to block the transmission of signals from the photoreceptors to the OFF-bipolar cells, horizontal cells, and other inner retinal neurons. Results:The amplitude of the b-wave of the photopic ERG was severely reduced in mGluR6-deficient mice, but a small, slow, positive component was seen after the a-wave. Intravitreous injection of PDA eliminated this positive component. Conclusions: The mGluR6-deficient mouse is a useful animal model to study the contribution of the ON-and OFF-pathways to the mouse ERG.

Type 4 OFF cone bipolar cells of the mouse retina express calsenilin and contact cones as well as rods

Immunocytochemical discrimination of distinct bipolar cell types in the mouse retina is a prerequisite for analyzing retinal circuitry in wild-type and transgenic mice. Here we demonstrate that among the more than 10 anatomically defined mouse bipolar cell types, type 4 bipolar cells are specifically recognized by anti-calsenilin antibodies. Axon terminals in the inner plexiform layer are not readily identifiable because calsenilin is also expressed in a subset of amacrine and ganglion cells. In contrast, in the outer plexiform layer calsenilin immunoreactivity allows the analysis of photoreceptor to type 4 bipolar cell contacts. A dense plexus of calsenilin-positive dendrites makes several basal contacts at cone pedicles. An individual calsenilin-positive bipolar cell contacts five to seven cones. In addition, some calsenilin-positive dendrites contact rod photoreceptors. On average we counted 10 rod spherule contacts per type 4 bipolar cell, and approximately 10% of rods contacted type 4 bipolar cells. We suggest that type 4 bipolar cells, together with the recently described type 3a and b cells, provide an alternative and direct route from rods to OFF cone bipolar cells. In the Bassoon DeltaEx4/5 mouse, a mouse mutant that shows extensive remodeling of the rod system including sprouting of horizontal and rod bipolar cells into the outer nuclear layer due to impaired synaptic transmission, we found that in addition mixed-input (type 3 and 4) OFF bipolar cells sprout to ectopic sites. In contrast, true cone-selective type 1 and 2 OFF cone bipolar cells did not show sprouting in the Bassoon mouse mutant.

Spatial Properties and Functional Organization of Small Bistratified Ganglion Cells in Primate Retina

The primate visual system consists of parallel pathways initiated by distinct cell types in the retina that encode different features of the visual scene. Small bistratified cells (SBCs), which form a major projection to the thalamus, exhibit blue-ON/yellow-OFF [S-ON/(L+M)-OFF] light responses thought to be important for high-acuity color vision. However, the spatial processing properties of individual SBCs and their spatial arrangement across the visual field are poorly understood. The present study of peripheral primate retina reveals that contrary to previous suggestions, SBCs exhibit center-surround spatial structure, with the (L+M)-OFF component of the receptive field approximately 50% larger in diameter than the S-ON component. Analysis of response kinetics shows that the (L+M)-OFF response in SBCs is slower than the S-ON response and significantly less transient than that of simultaneously recorded OFF-parasol cells. The (L+M)-OFF response in SBCs was eliminated by bath application of the metabotropic glutamate receptor agonist L-APB. These observations indicate that the (L+M)-OFF response of SBCs is not formed by OFF-bipolar cell input as has been suspected and suggest that it arises from horizontal cell feedback. Finally, the receptive fields of SBCs form orderly mosaics, with overlap and regularity similar to those of ON-parasol cells. Thus, despite their distinctive morphology and chromatic properties, SBCs exhibit two features of other retinal ganglion cell types: center-surround antagonism and regular mosaic sampling of visual space.

Islet-1 Controls the Differentiation of Retinal Bipolar and Cholinergic Amacrine Cells

Whereas the mammalian retina possesses a repertoire of factors known to establish general retinal cell types, these factors alone cannot explain the vast diversity of neuronal subtypes. In other CNS regions, the differentiation of diverse neuronal pools is governed by coordinately acting LIM-homeodomain proteins including the Islet-class factor Islet-1 (Isl1). We report that deletion of Isl1 profoundly disrupts retinal function as assessed by electroretinograms and vision as assessed by optomotor behavior. These deficits are coupled with marked reductions in mature ON- and OFF-bipolar (>76%), cholinergic amacrine (93%), and ganglion (71%) cells. Mosaic deletion of Isl1 permitted a chimeric analysis of "wild-type" cells in a predominantly Isl1-null environment, demonstrating a cell-autonomous role for Isl1 in rod bipolar and cholinergic amacrine development. Furthermore, the effects on bipolar cell development appear to be dissociable from the preceding retinal ganglion cell loss, because Pou4f2-null mice are devoid of similar defects in bipolar cell marker expression. Expression of the ON- and OFF-bipolar cell differentiation factors Bhlhb4 and Vsx1, respectively, requires the presence of Isl1, whereas the early bipolar cell marker Prox1 initially did not. Thus, Isl1 is required for engaging bipolar differentiation pathways but not for general bipolar cell specification. Spatiotemporal expression analysis of additional LIM-homeobox genes identifies a LIM-homeobox gene network during bipolar cell development that includes Lhx3 and Lhx4. We conclude that Isl1 has an indispensable role in retinal neuron differentiation within restricted cell populations and this function may reflect a broader role for other LIM-homeobox genes in retinal development, and perhaps in establishing neuronal subtypes.

Pharmacologically defined components of the normal porcine multifocal ERG

Multifocal electroretinograms (mfERG) from isoflurane anesthetized pigs were recorded and sequential application of TTX, NMDA, APB and PDA were used to identify contributions to the mfERG from inner retinal neurons, ON-pathway, OFF-pathway and photoreceptors. The cellular origins of the first-order kernel (K1) and the first slice of the second-order kernel (K2.1) porcine mfERG are contributed from both inner and outer retina. For the K1 waveform, the n1 involved responses of cone photoreceptors and OFF-bipolar cells. The leading edge of p1 is dominated by ON-bipolar cell depolarization. The rear edge of p1, n2 and p2 are dominated by ON-bipolar activities and shaped by the activities of OFF-bipolar cells and retinal cells with NMDAr and voltage-gated sodium channels other than ganglion cells. The p3 is mainly inner retinal activities. For the K2.1 waveform, the p1 and n1 are the summation of activities of ON-, OFF-bipolar cells and retinal cells rich in NMDAr and voltage-gated sodium channels other than ganglion cells. The p2 seems to be related to the ganglion cells. Better understanding of the cellular origins of the normal porcine mfERG will be useful for comparing and defining the functional changes that may occur in diseased retinas.

Neural Reprogramming in Retinal Degeneration

Early visual defects in degenerative diseases such as retinitis pigmentosa (RP) may arise from phased remodeling of the neural retina. The authors sought to explore the functional expression of ionotropic (iGluR) and group 3, type 6 metabotropic (mGluR6) glutamate receptors in late-stage photoreceptor degeneration. Excitation mapping with organic cations and computational molecular phenotyping were used to determine whether retinal neurons displayed functional glutamate receptor signaling in rodent models of retinal degeneration and a sample of human RP. After photoreceptor loss in rodent models of RP, bipolar cells lose mGluR6 and iGluR glutamate-activated currents, whereas amacrine and ganglion cells retain iGluR-mediated responsivity. Paradoxically, amacrine and ganglion cells show spontaneous iGluR signals in vivo even though bipolar cells lack glutamate-coupled depolarization mechanisms. Cone survival can rescue iGluR expression by OFF bipolar cells. In a case of human RP with cone sparing, iGluR signaling appeared intact, but the number of bipolar cells expressing functional iGluRs was double that of normal retina. RP triggers permanent loss of bipolar cell glutamate receptor expression, though spontaneous iGluR-mediated signaling by amacrine and ganglion cells implies that such truncated bipolar cells still release glutamate in response to some nonglutamatergic depolarization. Focal cone-sparing can preserve iGluR display by nearby bipolar cells, which may facilitate late RP photoreceptor transplantation attempts. An instance of human RP provides evidence that rod bipolar cell dendrite switching likely triggers new gene expression patterns and may impair cone pathway function.

Cellular Mechanisms for Direction Selectivity in the Retina

Direction selectivity represents a fundamental computation found across multiple sensory systems. In the mammalian visual system, direction selectivity appears first in the retina, where excitatory and inhibitory interneurons release neurotransmitter most rapidly during movement in a preferred direction. Two parallel sets of interneuron signals are integrated by a direction-selective ganglion cell, which creates a direction preference for both bright and dark moving objects. Direction selectivity of synaptic input becomes amplified by action potentials in the ganglion cell dendrites. Recent work has elucidated direction-selective mechanisms in inhibitory circuitry, but mechanisms in excitatory circuitry remain unexplained.

Synaptic influences on rat ganglion‐cell photoreceptors

The intrinsically photosensitive retinal ganglion cells (ipRGCs) provide a conduit through which rods and cones can access brain circuits mediating circadian entrainment, pupillary constriction and other non-image-forming visual functions. We characterized synaptic inputs to ipRGCs in rats using whole-cell and multielectrode array recording techniques. In constant darkness all ipRGCs received spontaneous excitatory and inhibitory synaptic inputs. Light stimulation evoked in all ipRGCs both synaptically driven ('extrinsic') and autonomous melanopsin-based ('intrinsic') responses. The extrinsic light responses were depolarizing, about 5 log units more sensitive than the intrinsic light response, and transient near threshold but sustained to brighter light. Pharmacological data showed that ON bipolar cells and amacrine cells make the most prominent direct contributions to these extrinsic light responses, whereas OFF bipolar cells make a very weak contribution. The spatial extent of the synaptically driven light responses was comparable to that of the intrinsic photoresponse, suggesting that synaptic contacts are made onto the entire dendritic field of the ipRGCs. These synaptic influences increase the sensitivity of ipRGCs to light, and also extend their temporal bandpass to higher frequencies. These extrinsic ipRGC light responses can explain some of the previously reported properties of circadian photoentrainment and other non-image-forming visual behaviours.

Retinal ganglion cell dendrites undergo a visual activity‐dependent redistribution after eye opening

Recent studies showed that light stimulation is required for the maturational segregation of retinal ganglion cell (RGC) synaptic connectivity with ON and OFF bipolar cells in mammalian retina. However, it is not clear to what extent light stimulation regulates the maturation of RGC dendritic ramification and synaptic connections. The present work quantitatively analyzed the dendritic ramification patterns of different morphological subtypes of RGCs of developing mouse retinas and demonstrated that RGCs in all four major morphological subtypes underwent profound dendritic redistributions from the center to specific stratum of the IPL after eye opening. Light deprivation preferentially blocked the developmental RGC dendritic redistribution from the center to sublamina a of the IPL. Interestingly, this developmental redistribution of RGC dendrites could not be explained by a simple developmental elimination of "excess" dendrites and, therefore, suggests a possible mechanism that requires both selective dendritic growth and elimination guided by visual activity.

Synaptotagmin I and II are present in distinct subsets of central synapses

Synaptotagmin 1 and 2 (syt 1, syt 2) are synaptic vesicle-associated membrane proteins that act as calcium sensors for fast neurotransmitter release from presynaptic nerve terminals. Here we show that widely used monoclonal antibodies, mab 48 and znp-1, stain nerve terminals in multiple species and, in mouse, recognize syt 1 and syt 2, respectively. With these antibodies, we examined the synaptic localization of these synaptotagmin isoforms in the mouse central nervous system. Syt 1 and syt 2 are localized predominantly to different subsets of synapses in retina, hippocampus, cerebellum, and median nucleus of the trapezoid body (MNTB). In the MNTB, syt 1 and syt 2 are present in different presynaptic terminals on the same postsynaptic principal neuron. In retina, horizontal and OFF-bipolar cell terminals contain syt 2, whereas most other terminals contain syt 1. Syt 1 localization in the immature retina resembles that seen in adult; however, syt 2 localization appears strikingly different at perinatal ages and continues to change dramatically prior to eye opening. For example, starburst amacrine cells, which lack syt 2 in adult retina, transiently express syt 2 during the first 2 postnatal weeks. In addition to differences in spatial and temporal distribution, species-specific differences in synaptotagmin localization were observed in retina and cerebellum. The cell-, temporal-, and species-specific expression of synaptotagmin isoforms suggests that each may have distinct functions in neurotransmitter release.

Cation Cl− cotransporters in the dendrites of goldfish bipolar cells

Bipolar cells receive an input from gamma-aminobutyric acid ergic horizontal cells in distal retinas. gamma-Aminobutyric acid can induce either an inhibitory or an excitatory Cl(-) response based on the intracellular Cl(-) concentration in the bipolar dendrites. Two cation Cl(-) cotransporters, Na-K-2Cl(-) and K-2Cl(-), differently control local Cl(-) levels in neurons by uptaking or extracting Cl(-), respectively. By using immunocytochemical techniques, we found that Na-K-2Cl(-) and K-2Cl(-) were preferentially distributed in On-bipolar and Off-bipolar dendrites in goldfish retinas. Na-K-2Cl(-) was colocalized with protein kinase C in On-bipolar dendrites, whereas K-2Cl(-) was located in Off-bipolar dendrites, fully overlapping with ionotropic glutamate receptor GluR4. Possibly, an internal Cl(-) level is higher in the On-bipolar dendrites than in the Off-bipolar dendrites. gamma-Aminobutyric acid inputs in the distal retina might excite On-bipolar cells, but inhibit Off-bipolar cells.

Connections of diffuse bipolar cells in primate retina are biased against S‐cones

In mammalian retina, each diffuse bipolar type stratifies in a distinct layer of the inner plexiform layer. Thus, different types of bipolar cells provide output to distinct visual pathways. Here, the question of whether diffuse bipolar cell types differ with respect to their contacts with short wavelength-sensitive (S-) cones was investigated in the retinas of a New World monkey, Callithrix jacchus, and an Old World monkey, Macaca fascicularis. Subpopulations of OFF bipolar cells were labeled with antibodies to the glutamate transporter Glt-1 and ON bipolar cells were labeled with antibodies to the alpha subunit of the Go protein (Goalpha). Two types of diffuse ON bipolar cells, DB4 and DB6, were identified with antibodies to protein kinase Calpha and CD15, respectively. Cone pedicles were labeled either with peanut agglutinin coupled to fluorescein or with antibodies to the ribbon protein, C-terminus binding protein 2. We found that immunoreactivity for Glt-1 (OFF bipolar cells) is reduced at S-cones in comparison to medium/long wavelength-sensitive (M/L-) cones. Immunoreactivity for Goalpha (ON bipolar cells) is comparable at all cone types. Nearly all M/L-cone pedicles contact the diffuse ON bipolar types DB4 and DB6, but only between 60% and 75% of the S-cone pedicles make contact. Furthermore, the number of dendritic tips of DB4 and DB6 cells at S-cone pedicles is lower than that at M/L-cone pedicles. These results suggest that there is a bias in the S-cone connectivity of diffuse bipolar cells.

Test of the paired-flash electroretinographic method in mice lacking b-waves

Previous studies of rod photoreceptors in vivo have employed a paired-flash electroretinographic (ERG) technique to determine rod response properties. To test whether absence versus presence of the ERG b-wave affects the photoreceptor response derived by the paired-flash method, we examined paired-flash-derived responses obtained from nob mice, a mutant strain with a defect in signal transduction between photoreceptors and ON bipolar cells that causes a lack of the b-wave. Normal littermates of the nob mice served as controls. The normalized amplitude-intensity relation of the derived response determined in nob mice at the near-peak time of 86 ms was similar to that determined for the controls. The full time course of the derived rod response was obtained for test flash strengths ranging from 0.11 to 17.38 scotopic cd s m(-2) (sc cd s m(-2)). Time-course data obtained from nob and control mice exhibited significant but generally modest differences. With saturating test flash strengths, half-recovery times for the derived response of nob versus control mice differed by approximately 60 ms or less about the combined (nob and control) average respective values. Time course data also were obtained before versus after intravitreal injection of L-2-amino-4-phosphonobutyrate (APB) (which blocks transmission from photoreceptors to depolarizing bipolar cells) and of cis 2,3-piperidine dicarboxylic acid (PDA) (which blocks transmission to OFF bipolar cells, and to horizontal, amacrine and ganglion cells). Neither APB nor PDA substantially affected derived responses obtained from nob or control mice. The results provide quantitative information on the effect of b-wave removal on the paired-flash-derived response in mouse. They argue against a substantial skewing effect of the b-wave on the paired-flash-derived response obtained in normal mice and are consistent with the notion that, to good approximation, this derived response represents the isolated flash response of the photoreceptors in both nob and normal mice.

Experimental hypoxia in human eyes: Implications for ischaemic disease

Objective This study investigated neuroretinal activity under normoxic and hypoxic conditions with the multifocal electroretinogram (mfERG). Methods We used two mfERG paradigms, the fast flicker and slow flash stimulation modes, to measure neuroretinal activity in five healthy participants who breathed room air and a reduced oxygen mixture (14% oxygen, balance nitrogen). We analysed concentric ring N1P1 and P1N2 response density amplitudes, the P1 implicit times as well as the local scalar product (SP) response densities. Results During hypoxia there was a significant reduction of the scalar product response density for the fast flicker ( p < 0.001) and for the slow flash mfERG ( p < 0.001). The N1P1 and P1N2 response densities were lower especially for the central three rings; although these reductions were not significant between the two oxygen conditions, they indicated an overall distortion of the mfERG waveform. Conclusions It is demonstrated that a postreceptoral, primarily ON and OFF bipolar cell deficit is evident in the central retina of healthy young people during short term hypoxia. Significance Our findings suggest that persons with pre-existing ischaemic eye disease may be at risk when exposed to hypoxic conditions.

Cross-talk between ON and OFF channels in the salamander retina: Indirect bipolar cell inputs to ON–OFF ganglion cells

It has been widely accepted that ON and OFF channels in the visual system are segregated with little cross-communication, except for the mammalian rod bipolar cell-AII amacrine cell-ganglion cell pathway. Here, we show that in the tiger salamander retina the light responses of a subpopulation of ON–OFF ganglion cells are mediated by crossing the ON and OFF bipolar cell pathways. Although the majority of ON–OFF ganglion cells (type I cells) receive direct excitatory inputs from depolarizing and hyperpolarizing bipolar cells (DBCs and HBCs), about 5% (type II cells) receive indirect excitatory inputs from DBCs and 20% (type III cells) receive indirect excitatory inputs from HBCs. These indirect bipolar cell inputs are likely to be mediated by a subpopulation of amacrine cells that exhibit transient hyperpolarizing light responses (AC Hs) and make GABAergic/glycinergic synapses on DBC or HBC axon terminals. GABA and glycine receptor antagonists enhanced the ON and OFF excitatory cation current (Δ I C) in type I ganglion cells, but completely suppressed the ON Δ I C mediated by DBCs in type II cells and the OFF Δ I C mediated by HBCs in types III cells. Dendrites of type I cells ramify in both sublamina A and B, type II cells exclusively in sublamina A, and type III cells exclusively in sublamina B of the inner plexiform layer. These results demonstrate that indirect, amacrine cell-mediated bipolar cell-ganglion cell synaptic pathways exist in a non-mammalian retina, and that bidirectional cross-talk between ON and OFF channels is present in the vertebrate retina.

G protein coupling profile of mGluR6 and expression of Gα proteins in retinal ON bipolar cells

Metabotropic glutamate receptor 6 (mGluR6) is a group III, pertussis toxin (PTX)-sensitive G protein coupled mGluR that plays a specialized role in the retina. Retinal ON bipolar cells, which receive direct glutamatergic input from photoreceptor cells, express mGluR6 as their primary postsynaptic glutamate receptor. Activation of mGluR6 in these cells initiates an intracellular signaling cascade ultimately leading to inhibition of a cation channel and cell hyperpolarization. The primary mediator of this pathway in vivo is G alpha(o), but the potential roles of other G proteins from the G alpha(i/o) family in the regulation of this or other signaling pathways in ON bipolar cells are unclear. To determine which specific G proteins from the G alpha(i/o) family are able to couple to mGluR6, a G alpha reconstitution system was employed using PTX-insensitive G alpha mutants expressed with mGluR6 in PTX-treated sympathetic neurons from the rat superior cervical ganglion (SCG). The efficiency of coupling to mGluR6 was G(oa) > G(ob), G(i1) > G(i2), G(i3), whereas no coupling was observed with G alpha(z), nor with the retinal G alpha proteins, rod (GNAT2) or cone (GNAT1) transducin (G alpha(Tr-R), G alpha(Tr-C)). Finally, the expression of G alpha proteins determined to couple with mGluR6 was examined in rat ON bipolar cells using single cell RT-PCR. Co-expression of mGluR6 message was used to distinguish ON from OFF bipolar cells. Expression of G alpha(o) was detected in every ON bipolar cell examined. Message for G alpha(i1), which coupled moderately to mGluR6, was not detected in ON bipolar cells, nor was G alpha(i3), which coupled to mGluR6 in only a few cells but on average did not exhibit statistically significant coupling. Finally, though G alpha(i2) was detectable in ON bipolar cells, its coupling to mGluR6 in the SCG system was not significant. Together, these data indicate that signaling through mGluR6 in mammalian ON bipolar cells is highly focused, apparently acting through a single G alpha protein subtype.

Slow flash multifocal electroretinogram in myopia

Purpose The purpose of this study was to investigate the characteristics of retinal function in myopes using a modified multifocal electroretinogram (mfERG) protocol, the slow flash (sf-mfERG) paradigm, which is thought to primarily reflect responses of ON- and OFF-bipolar cells and emphasize the late response components. Methods Twenty-eight subjects (10 emmetropes and 18 myopes) underwent mfERG testing using VERIS 5.1.5X. The sf-mfERG stimulus array consisted of 103-scaled hexagons and flickered according to a pseudorandom binary m-sequence (2 13-1). The stimulation sequence was slowed by inserting three dark frames such that each step in the m-sequence was four frames long (53.3 ms). The amplitude and implicit time of the major sf-mfERG waveform features (N1, P1, and N2) of the first-order kernel were analysed. Results Myopes had significantly reduced P1 and N2 amplitudes compared to the emmetropes ( F 1, 25 = 8.818, p = 0.007; F 1, 25 = 6.723, p = 0.017). There were no significant differences in N1 amplitude or implicit time between the groups ( F 1, 25 = 1.506, p = 0.233; F 1, 25 = 1.291, p = 0.269). Conclusions Late response components (P1 and N2) of the first-order sf-mfERG responses were preferentially affected in myopia, suggesting possible reduced ON- and OFF-bipolar cell activity. As bipolar cells form the first synapse of the visual system with the photoreceptors to initiate the ON- and OFF-pathways, future investigations of ON- and OFF-systems in myopia are of interest.