OFF Responses Research Articles

Melanopsin-driven increases in maintained activity enhance thalamic visual response reliability across a simulated dawn

Twice a day, at dawn and dusk, we experience gradual but very high amplitude changes in background light intensity (irradiance). Although we perceive the associated change in environmental brightness, the representation of such very slow alterations in irradiance by the early visual system has been little studied. Here, we addressed this deficit by recording electrophysiological activity in the mouse dorsal lateral geniculate nucleus under exposure to a simulated dawn. As irradiance increased we found a widespread enhancement in baseline firing that extended to units with ON as well as OFF responses to fast luminance increments. This change in baseline firing was equally apparent when the slow irradiance ramp appeared alone or when a variety of higher-frequency artificial or natural visual stimuli were superimposed upon it. Using a combination of conventional knockout, chemogenetic, and receptor-silent substitution manipulations, we continued to show that, over higher irradiances, this increase in firing originates with inner-retinal melanopsin photoreception. At the single-unit level, irradiance-dependent increases in baseline firing were strongly correlated with improvements in the amplitude of responses to higher-frequency visual stimuli. This in turn results in an up to threefold increase in single-trial reliability of fast visual responses. In this way, our data indicate that melanopsin drives a generalized increase in dorsal lateral geniculate nucleus excitability as dawn progresses that both conveys information about changing background light intensity and increases the signal:noise for fast visual responses.

Solid state fluorescent functionalized-triphenylamine Bodipy detector for HCl vapor with high stability and absolute fluorescent quantum yield

A series of three new boron-dipyrromethenes bringing different functionalized triphenylamine donor Bodipy–TPA–R (R = BI, BH–OH, BH–COOH) (1–3) together with the reference compound Bodipy–TPA (0), have been synthesized and structurally characterized. Due to their same less electronic coupling between the triphenylamine donor and Bodipy acceptor in ground state and in particular the twisted intramolecular charge transfer (TICT) between them in excited state, these functionalized Bodipy derivatives exhibit similar optical properties in solution but not in the solid state owing to their differently intermolecular interaction and molecular packing associated with the diverse triphenylamine-modification. The lack of effective face-to-face π–π stacking interaction between Bodipy luminophores in crystalline states of 0 and 3, in combination with the restriction of the twisting motion and charge transfer between their triphenylamine moiety and Bodipy framework, results in the obvious aggregation- (AIE) and crystallization-induced emission (CIE) effect, with the absolute fluorescent quantum yield (Φ) as high as 19.47% and 12.30% for single crystals of 0 and 3, respectively. In good contrast, despite the same complete restriction of the functionalized triphenylamine donor around Bodipy acceptor, the presence of partial overlapping face-to-face π–π interaction between two parallel C9BN2 fluorophores leads to very low Φ being 1.37% and 3.40% for the single crystals of 1 and 2 as well as the negligible AIE effect. Moreover, the CIE-active red-emission for 3 can be switched between bright and dark states along with the change from the crystalline state to amorphous phase by heating–cooling process. Notably, functionalization over triphenylamine moiety with carboxyl-benzoylhydrazone group in Bodipy–TPA–BH–COOH (3) renders this compound a good solid-state fluorescent sensor due to its fast fluorescence ON–OFF response for HCl vapor in thin film.

Functional segregation of retinal ganglion cell projections to the optic tectum of rainbow trout.

The interpretation of visual information relies on precise maps of retinal representation in the brain coupled with local circuitry that encodes specific features of the visual scenery. In nonmammalian vertebrates, the main target of ganglion cell projections is the optic tectum. Although the topography of retinotectal projections has been documented for several species, the spatiotemporal patterns of activity and how these depend on background adaptation have not been explored. In this study, we used a combination of electrical and optical recordings to reveal a retinotectal map of ganglion cell projections to the optic tectum of rainbow trout and characterized the spatial and chromatic distribution of ganglion cell fibers coding for increments (ON) and decrements (OFF) of light. Recordings of optic nerve activity under various adapting light backgrounds, which isolated the input of different cone mechanisms, yielded dynamic patterns of ON and OFF input characterized by segregation of these two fiber types. Chromatic adaptation decreased the sensitivity and response latency of affected cone mechanisms, revealing their variable contributions to the ON and OFF responses. Our experiments further demonstrated restricted input from a UV cone mechanism to the anterolateral optic tectum, in accordance with the limited presence of UV cones in the dorsotemporal retina of juvenile rainbow trout. Together, our findings show that retinal inputs to the optic tectum of this species are not homogeneous, exhibit highly dynamic activity patterns, and are likely determined by a combination of biased projections and specific retinal cell distributions and their activity states.

Morphology and function of three VIP-expressing amacrine cell types in the mouse retina.

Amacrine cells (ACs) are the most diverse class of neurons in the retina. The variety of signals provided by ACs allows the retina to encode a wide range of visual features. Of the 30-50 AC types in mammalian species, few have been studied in detail. Here, we combine genetic and viral strategies to identify and to characterize morphologically three vasoactive intestinal polypeptide-expressing GABAergic AC types (VIP1-, VIP2-, and VIP3-ACs) in mice. Somata of VIP1- and VIP2-ACs reside in the inner nuclear layer and somata of VIP3-ACs in the ganglion cell layer, and they show asymmetric distributions along the dorsoventral axis of the retina. Neurite arbors of VIP-ACs differ in size (VIP1-ACs ≈ VIP3-ACs > VIP2-ACs) and stratify in distinct sublaminae of the inner plexiform layer. To analyze light responses and underlying synaptic inputs, we target VIP-ACs under 2-photon guidance for patch-clamp recordings. VIP1-ACs depolarize strongly to light increments (ON) over a wide range of stimulus sizes but show size-selective responses to light decrements (OFF), depolarizing to small and hyperpolarizing to large stimuli. The switch in polarity of OFF responses is caused by pre- and postsynaptic surround inhibition. VIP2- and VIP3-ACs both show small depolarizations to ON stimuli and large hyperpolarizations to OFF stimuli but differ in their spatial response profiles. Depolarizations are caused by ON excitation outweighing ON inhibition, whereas hyperpolarizations result from pre- and postsynaptic OFF-ON crossover inhibition. VIP1-, VIP2-, and VIP3-ACs thus differ in response polarity and spatial tuning and contribute to the diversity of inhibitory and neuromodulatory signals in the retina.

Proton block of proton-activated TRPV1 current.

The TRPV1 cation channel is a polymodal nociceptor that is activated by heat and ligands such as capsaicin and is highly sensitive to changes in extracellular pH. In the body core, where temperature is usually stable and capsaicin is normally absent, H(+) released in response to ischemia, tissue injury, or inflammation is the best-known endogenous TRPV1 agonist, activating the channel to mediate pain and vasodilation. Paradoxically, removal of H(+) elicits a transient increase in TRPV1 current that is much larger than the initial H(+)-activated current. We found that this prominent OFF response is caused by rapid recovery from H(+) inhibition of the excitatory current carried by H(+)-activated TRPV1 channels. H(+) inhibited current by interfering with ion permeation. The degree of inhibition is voltage and permeant ion dependent, and it can be affected but not eliminated by mutations to acidic residues within or near the ion selectivity filter. The opposing H(+)-mediated gating and permeation effects produce complex current responses under different cellular conditions that are expected to greatly affect the response of nociceptive neurons and other TRPV1-expressing cells.

Temperature representation in the Drosophila brain.

In Drosophila, rapid temperature changes are detected at the periphery by dedicated receptors forming a simple sensory map for hot and cold in the brain. However, flies show a host of complex innate and learned responses to temperature, indicating that they are able to extract a range of information from this simple input. Here we define the anatomical and physiological repertoire for temperature representation in the Drosophila brain. First, we use a photolabelling strategy to trace the connections that relay peripheral thermosensory information to higher brain centres, and show that they largely converge onto three target regions: the mushroom body, the lateral horn (both of which are well known centres for sensory processing) and the posterior lateral protocerebrum, a region we now define as a major site of thermosensory representation. Next, using in vivo calcium imaging, we describe the thermosensory projection neurons selectively activated by hot or cold stimuli. Fast-adapting neurons display transient ON and OFF responses and track rapid temperature shifts remarkably well, while slow-adapting cell responses better reflect the magnitude of simple thermal changes. Unexpectedly, we also find a population of broadly tuned cells that respond to both heating and cooling, and show that they are required for normal behavioural avoidance of both hot and cold in a simple two-choice temperature preference assay. Taken together, our results uncover a coordinated ensemble of neural responses to temperature in the Drosophila brain, demonstrate that a broadly tuned thermal line contributes to rapid avoidance behaviour, and illustrate how stimulus quality, temporal structure, and intensity can be extracted from a simple glomerular map at a single synaptic station.

Visual homeostatic processing in V1: when probability meets dynamics.

Visual homeostatic processing in V1: when probability meets dynamics.

The electroretinogram of Mongolian gerbil (Meriones unguiculatus): Comparison to mouse

The Mongolian gerbil (Meriones unguiculatus) is a diurnal rodent whose retinal photoreceptors comprise 13% cones in contrast to 1–3% in nocturnal mice and rats. Moreover, it displays a retinal structure more analogous to that of human than of mouse. However, the electroretinogram (ERG) recordings of gerbils have not yet been well studied. Thus, here we compared the ERGs of gerbils and C57 mice. We recorded responses to full-field flashes of increasing intensities under both dark and light adaptation. We also investigated responses to flickers of increasing frequencies and to long-duration flashes under photopic conditions. In scotopic, the amplitudes of the gerbil a- and b-waves are slightly smaller than those of the mouse waves. However, in photopic, the gerbil wave amplitudes are 2-fold larger than those of mice. Gerbils also exhibit larger flicker responses and higher flicker fusion frequencies than mice. Furthermore, unlike mice, gerbils show a positive OFF response (d-wave) and a post b-wave positive potential (i-wave), features commonly observed in human photopic ERGs. Our results suggest that gerbils may complement rod-dominant mice as models for studying retinal cone function and pathologies.

Opposite effects of ethanol on electroretinographic ON and OFF responses are due to its action in the distal retina

Abstract The aim of the present study was to give insight into the mechanisms of action of ethanol on the function of retinal ON and OFF channels. In order to fulfill this aim the effects of ethanol on the electroretinographic (ERG) ON

Targeting Channelrhodopsin-2 to ON-bipolar Cells With Vitreally Administered AAV Restores ON and OFF Visual Responses in Blind Mice

Most inherited retinal dystrophies display progressive photoreceptor cell degeneration leading to severe visual impairment. Optogenetic reactivation of retinal neurons mediated by adeno-associated virus (AAV) gene therapy has the potential to restore vision regardless of patient-specific mutations. The challenge for clinical translatability is to restore a vision as close to natural vision as possible, while using a surgically safe delivery route for the fragile degenerated retina. To preserve the visual processing of the inner retina, we targeted ON bipolar cells, which are still present at late stages of disease. For safe gene delivery, we used a recently engineered AAV variant that can transduce the bipolar cells after injection into the eye's easily accessible vitreous humor. We show that AAV encoding channelrhodopsin under the ON bipolar cell-specific promoter mediates long-term gene delivery restricted to ON-bipolar cells after intravitreal administration. Channelrhodopsin expression in ON bipolar cells leads to restoration of ON and OFF responses at the retinal and cortical levels. Moreover, light-induced locomotory behavior is restored in treated blind mice. Our results support the clinical relevance of a minimally invasive AAV-mediated optogenetic therapy for visual restoration.

Proton as a Dual Regulator for TRPV1

TRPV1 is a pain-sensing polymodal receptor. In the body core where temperature is stable and capsaicin is normally absent, extracellular H+ (from tissue injury, inflammation, ischemia, etc.) serves as the best-known endogenous ligand. While H+ is known to strongly activate TRPV1, the underlying mechanism is still unclear. We observe two opposing effects of H+: it strongly potentiates activation gating but inhibits ion permeation. The combination of these two effects produces a smaller current upon acidification and a prominent OFF response when H+ is removed. The permeation effect is voltage-dependent, indicating that H+ binds in the conducting pore to cause block. Correlations between TRPV1 cryo-EM structures and functional effects of point mutations allow us to investigate the mechanism underlying H+-mediated permeation block. Inhibition of ion permeation substantially shifts the macroscopic dose-response relationship for H+ activation, which needs to be corrected for the study of gating effects.

Molecular Mechanism of TRPV1 Activation by Capsaicin

Molecular Mechanism of TRPV1 Activation by Capsaicin

ON-OFF Interactions in the Retina: Role of Glycine and GABA.

In the vertebrate retina, visual signals are segregated into parallel ON and OFF pathways, which provide information for light increments and decrements. The segregation is first evident at the level of the ON and OFF bipolar cells and it apparently remains as signals propagate to higher brain visual centers. A fundamental question in visual neuroscience is how these two parallel pathways function: are they independent from each other or do they interact somehow? In the latter case, what kinds of mechanisms are involved and what are the consequences from this cross-talk? This review summarizes current knowledge about the types of interactions between the ON and OFF channels in nonmammalian and mammalian retina. Data concerning the ON-OFF interactions in distal retina revealed by recording of single bipolar cell activity and electroretinographic ON (b-wave) and OFF (d-wave) responses are presented. Special emphasis is put on the ON-OFF interactions in proximal retina and their dependence on the state of light adaptation in mammalian retina. The involvement of the GABAergic and glycinergic systems in the ON-OFF crosstalk is also discussed.

Effects of dopamine on response properties of ON-OFF RGCs in encoding stimulus durations

Single retinal ganglion cell's (RGCs) response properties, such as spike count and response latency, are known to encode some features of visual stimuli. On the other hand, neuronal response can be modulated by dopamine (DA), an important endogenous neuromodulator in the retina. In the present study, we investigated the effects of DA on the spike count and the response latency of bullfrog ON-OFF RGCs during exposure to different stimulus durations. We found that neuronal spike count and response latency were both changed with stimulus durations, and exogenous DA (10 μM) obviously attenuated the stimulus-duration-dependent response latency change. Information analysis showed that the information about light ON duration was mainly carried by the OFF response and vice versa, and the stimulation information was carried by both spike count and response latency. However, during DA application, the information carried by the response latency was greatly decreased, which suggests that dopaminergic pathway is involved in modulating the role of response latency in encoding the information about stimulus durations.

Disinhibitory recruitment of NMDA receptor pathways in retina

Glutamate release at bipolar to ganglion cell synapses activates NMDA and AMPA/kainic acid (KA) ionotropic glutamate receptors. Their relative strength determines the output signals of the retina. We found that this balance is tightly regulated by presynaptic inhibition that preferentially suppresses NMDA receptor (NMDAR) activation. In transient ON-OFF neurons, block of GABA and glycine feedback enhanced total NMDAR charge by 35-fold in the ON response and 9-fold in the OFF compared with a 1.7-fold enhancement of AMPA/KA receptors. Blocking only glycine receptors enhanced the NMDAR excitatory postsynaptic current 10-fold in the ON and 2-fold in the OFF pathway. Blocking GABA(A) or GABA(C) receptors (GABA(C)Rs or GABA(A)Rs) produced small changes in total NMDAR charge. When both GABA(A)Rs and GABA(C)Rs were blocked, the total NMDAR charge increased ninefold in the ON and fivefold in the OFF pathway. This exposed a strong GABA(C)R feedback to bipolar cells that was suppressed by serial amacrine cell synapses mediated by GABA(A)Rs. The results indicate that NMDAR currents are large but latent, held in check by dual GABA and glycine presynaptic inhibition. One example of this controlled NMDAR activation is the cross talk between ON and OFF pathways. Blocking the ON pathway increased NMDAR relative strength in the OFF pathway. Stimulus prolongation similarly increased the NMDAR relative strength in the OFF response. This NMDAR enhancement was produced by a diminution in GABA and glycine feedback. Thus the retinal network recruits NMDAR pathways through presynaptic disinhibition.

Neuronal nonlinearity explains greater visual spatial resolution for darks than lights

Astronomers and physicists noticed centuries ago that visual spatial resolution is higher for dark than light stimuli, but the neuronal mechanisms for this perceptual asymmetry remain unknown. Here we demonstrate that the asymmetry is caused by a neuronal nonlinearity in the early visual pathway. We show that neurons driven by darks (OFF neurons) increase their responses roughly linearly with luminance decrements, independent of the background luminance. However, neurons driven by lights (ON neurons) saturate their responses with small increases in luminance and need bright backgrounds to approach the linearity of OFF neurons. We show that, as a consequence of this difference in linearity, receptive fields are larger in ON than OFF thalamic neurons, and cortical neurons are more strongly driven by darks than lights at low spatial frequencies. This ON/OFF asymmetry in linearity could be demonstrated in the visual cortex of cats, monkeys, and humans and in the cat visual thalamus. Furthermore, in the cat visual thalamus, we show that the neuronal nonlinearity is present at the ON receptive field center of ON-center neurons and ON receptive field surround of OFF-center neurons, suggesting an origin at the level of the photoreceptor. These results demonstrate a fundamental difference in visual processing between ON and OFF channels and reveal a competitive advantage for OFF neurons over ON neurons at low spatial frequencies, which could be important during cortical development when retinal images are blurred by immature optics in infant eyes.

Response dynamics of bullfrog ON-OFF RGCs to different stimulus durations

Stimulus duration is an important feature of visual stimulation. In the present study, response properties of bullfrog ON-OFF retinal ganglion cells (RGCs) in exposure to different visual stimulus durations were studied. By using a multi-electrode recording system, spike discharges from ON-OFF RGCs were simultaneously recorded, and the cells' ON and OFF responses were analyzed. It was found that the ON response characteristics, including response latency, spike count, as well as correlated activity and relative latency between pair-wise cells, were modulated by different light OFF intervals, while the OFF response characteristics were modulated by different light ON durations. Stimulus information carried by the ON and OFF responses was then analyzed, and it was found that information about different light ON durations was more carried by transient OFF response, whereas information about different light OFF intervals were more carried by transient ON response. Meanwhile, more than 80% information about stimulus durations was carried by firing rate. These results suggest that ON-OFF RGCs are sensitive to different stimulus durations, and they can efficiently encode the information about visual stimulus duration by firing rate.

Ionotropic GABA Receptors and Distal Retinal ON and OFF Responses.

In the vertebrate retina, visual signals are segregated into parallel ON and OFF pathways, which provide information for light increments and decrements. The segregation is first evident at the level of the ON and OFF bipolar cells in distal retina. The activity of large populations of ON and OFF bipolar cells is reflected in the b- and d-waves of the diffuse electroretinogram (ERG). The role of gamma-aminobutyric acid (GABA), acting through ionotropic GABA receptors in shaping the ON and OFF responses in distal retina, is a matter of debate. This review summarized current knowledge about the types of the GABAergic neurons and ionotropic GABA receptors in the retina as well as the effects of GABA and specific GABAA and GABAC receptor antagonists on the activity of the ON and OFF bipolar cells in both nonmammalian and mammalian retina. Special emphasis is put on the effects on b- and d-waves of the ERG as a useful tool for assessment of the overall function of distal retinal ON and OFF channels. The role of GABAergic system in establishing the ON-OFF asymmetry concerning the time course and absolute and relative sensitivity of the ERG responses under different conditions of light adaptation in amphibian retina is also discussed.

Effect of GABAergic inhibition on odorant concentration coding in mushroom body intrinsic neurons of the honeybee

Kenyon cells, the intrinsic neurons of the insect mushroom body, have the intriguing property of responding in a sparse way to odorants. Sparse neuronal codes are often invariant to changes in stimulus intensity and duration, and sparse coding often depends on global inhibition. We tested if this is the case for honeybees' Kenyon cells, too, and used in vivo Ca²⁺ imaging to record their responses to different odorant concentrations. Kenyon cells responded not only to the onset of odorant stimuli (ON responses), but also to their termination (OFF responses). Both, ON and OFF responses increased with increasing odorant concentration. ON responses were phasic and invariant to the duration of odorant stimuli, while OFF responses increased with increasing odorant duration. Pharmacological blocking of GABA receptors in the brain revealed that ionotropic GABA(A) and metabotropic GABA(B) receptors attenuate Kenyon cells' ON responses without changing their OFF responses. Ionotropic GABA(A) receptors attenuated Kenyon cell ON responses more strongly than metabotropic GABA(B) receptors. However, the response dynamic, temporal resolution and paired-pulse depression did not depend on GABA(A) transmission. These data are discussed in the context of mechanisms leading to sparse coding in Kenyon cells.

Electroretinographic assessment of retinal function during acute exposure to normobaric hypoxia

The current study aimed to investigate retinal function during exposure to normobaric hypoxia. Standard Ganzfeld ERG equipment (Diagnosys LLC, Cambridge, UK) using an extended ISCEV protocol was applied to explore intensity-response relationship in dark- and light- adapted conditions in 13 healthy volunteers (mean age 25 ± 3years). Baseline examinations were performed under atmospheric air conditions at 341meters above sea level (FIO2 of 21%), and were compared to hypoxia (FIO2 of 13.2%) by breathing a nitrogen-enriched gas mixture for 45min. All subjects were monitored using infrared oximetry and blood gas analysis. The levels of PaCO2 changed from 38.4 ± 2.7mmHg to 36.4 ± 3.0mmHg, PaO2 from 95.5 ± 1.9mmHg to 83.7 ± 4.6mmHg, and SpO2 from 100 ± 0% to 87 ± 4%, from baseline to hypoxia respectively. A significant decrease (p < 0.05) was found for saturation amplitude of the dark-adapted b-wave intensity-response function (Vmax), dark-adapted a- and b-wave amplitudes of combined rod and cone responses (3 and 10cd.s/m(2)), light-adapted b-wave amplitudes of single flash (3 and 10cd.s/m(2)), and flicker responses (5-45Hz) during hypoxia compared to baseline, without changes in implicit times. The a-wave slope of combined rod and cone responses (3 and 10cd.s/m(2)) and the oscillatory potentials were significantly lower during hypoxia (p < 0.05). A isolated light-adapted ON response (250ms flash) showed a reduction of amplitudes at hypoxia (p < 0.05), but no changes were observed for the OFF response. The results show significant impairment of retinal function during simulated normobaric short-term hypoxia affecting specific retinal cells of rod and cone pathways.