Dopamine differentially affects retinal circuits to shape the retinal code.

Dopamine has long been reported to enhance antagonistic surrounds of retinal ganglion cells (RGCs). Yet, the retina contains many different RGC subtypes and the effects of dopamine can be subtype-specific. Using multielectrode array (MEA) recordings we investigated how dopamine shapes the receptive fields of RGCs in the mouse retina. We found that the non-selective dopamine receptor agonist apomorphine can either increase or decrease RGCs' surround strength, depending on their subtype. We then used two-photon targeted patch-clamp to target a specific RGC subtype, the transient-Off-αRGC. In line with our MEA recordings, apomorphine did not increase the antagonistic surround of transient-Off-αRGCs but enhanced their responses to Off stimuli in the centre receptive field. Both D1 - and D2 -like family receptor (D1 -R and D2 -R) blockers had the opposite effect and reduced centre-mediated responses, but differently affected transient-Off-αRGC's surround. While D2 -R blocker reduced surround antagonism, D1 -R blocker led to surround activation, revealing On responses to large stimuli. Using voltage-clamp recordings we separated excitatory inputs from Off cone bipolar cells and inhibitory inputs from the primary rod pathway. In control conditions, cone inputs displayed strong surround antagonism, while inputs from the primary rod pathway showed no surround. Yet, the surround activation in the D1 -R blockade originated from the primary rod pathway. Our findings demonstrate that dopamine differentially affects RGC subtypes via distinct pathways, suggesting that dopamine has a more complex role in shaping the retinal code than previously reported. KEY POINTS: Receptive fields of retinal ganglion cells (RGCs) have a centre-surround organisation, and previous work has shown that this organisation can be modulated by dopamine in a light-intensity-dependent manner. Dopamine is thought to enhance RGCs' antagonistic surround, but a detailed understanding of how different RGC subtypes are affected is missing. Using a multielectrode array recordings, clustering analysis and pharmacological manipulations, we found that dopamine can either enhance or weaken antagonistic surrounds, and also change response kinetics, of RGCs in a subtype-specific manner. We performed targeted patch-clamp recordings of one RGC subtype, the transient-Off-αRGC, and identified the underlying circuits by which dopamine shapes its receptive field. Our findings demonstrate that dopamine acts in a subtype-specific manner and can have complex effects, which has implications for other retinal computations that rely on receptive field structure.

Realistic retinal modeling unravels the differential role of excitation and inhibition to starburst amacrine cells in direction selectivity.

Retinal direction-selectivity originates in starburst amacrine cells (SACs), which display a centrifugal preference, responding with greater depolarization to a stimulus expanding from soma to dendrites than to a collapsing stimulus. Various mechanisms were hypothesized to underlie SAC centrifugal preference, but dissociating them is experimentally challenging and the mechanisms remain debatable. To address this issue, we developed the Retinal Stimulation Modeling Environment (RSME), a multifaceted data-driven retinal model that encompasses detailed neuronal morphology and biophysical properties, retina-tailored connectivity scheme and visual input. Using a genetic algorithm, we demonstrated that spatiotemporally diverse excitatory inputs-sustained in the proximal and transient in the distal processes-are sufficient to generate experimentally validated centrifugal preference in a single SAC. Reversing these input kinetics did not produce any centrifugal-preferring SAC. We then explored the contribution of SAC-SAC inhibitory connections in establishing the centrifugal preference. SAC inhibitory network enhanced the centrifugal preference, but failed to generate it in its absence. Embedding a direction selective ganglion cell (DSGC) in a SAC network showed that the known SAC-DSGC asymmetric connectivity by itself produces direction selectivity. Still, this selectivity is sharpened in a centrifugal-preferring SAC network. Finally, we use RSME to demonstrate the contribution of SAC-SAC inhibitory connections in mediating direction selectivity and recapitulate recent experimental findings. Thus, using RSME, we obtained a mechanistic understanding of SACs' centrifugal preference and its contribution to direction selectivity.

Transgenic mice reveal unexpected diversity of on-off direction-selective retinal ganglion cell subtypes and brain structures involved in motion processing.

On-Off direction-selective retinal ganglion cells (DSGCs) encode the axis of visual motion. They respond strongly to an object moving in a preferred direction and weakly to an object moving in the opposite, "null," direction. Historically, On-Off DSGCs were classified into four subtypes according to their directional preference (anterior, posterior, superior, or inferior). Here, we compare two genetically identified populations of On-Off DSGCs: dopamine receptor 4 (DRD4)-DSGCs and thyrotropin-releasing hormone receptor (TRHR)-DSGCs. We find that although both populations are tuned for posterior motion, they can be distinguished by a variety of physiological and anatomical criteria. First, the directional tuning of TRHR-DSGCs is broader than that of DRD4-DSGCs. Second, whereas both populations project similarly to the dorsal lateral geniculate nucleus, they project differently to the ventral lateral geniculate nucleus and the superior colliculus. Moreover, TRHR-DSGCs, but not DRD4-DSGCs, also project to the zona incerta, a thalamic area not previously known to receive direction-tuned visual information. Our findings reveal unexpected diversity among mouse On-Off DSGC subtypes that uniquely process and convey image motion to the brain.

Neurons in both pallidal segments change their firing properties similarly prior to closure of the eyes.

Current anatomical models of the cortico-basal ganglia (BG) network predict reciprocal discharge patterns between the external and internal segments of the globus pallidus (GPe and GPi, respectively), as well as cortical driving of BG activity. However, physiological studies revealing similarity in the transient responses of GPe and GPi neurons cast doubts on these predictions. Here, we studied the discharge properties of GPe, GPi, and primary motor cortex neurons of two monkeys in two distinct states: when eyes are open versus when they are closed. Both pallidal populations exhibited decreased discharge rates in the "eye closed" state accompanied by elevated values of the coefficient of variation (CV) of their interspike interval (ISI) distributions. The pallidal modulations in discharge patterns were partially attributable to larger fractions of longer ISIs in the "eye closed" state. In addition, the pallidal discharge modulations were gradual, starting prior to closing of the eyes. Cortical neurons, as opposed to pallidal neurons, increased their discharge rates steeply on closure of the eyes. Surprisingly, the cortical rate modulations occurred after pallidal modulations. However, as in the pallidum, the CV values of cortical ISI distributions increased in the "eye closed" state, indicating a more bursty discharge pattern in that state. Thus changes in GPe and GPi discharge properties were positively correlated, suggesting that the subthalamic nucleus and/or the striatum constitute the main common driving force for both pallidal segments. Furthermore, the early, unexpected changes in the pallidum are better explained by a subcortical rather than a cortical loop through the BG.

Topographic Variations in Retinal Encoding of Visual Space.

A retina completely devoid of topographic variations would be homogenous, encoding any given feature uniformly across the visual field. In a naive view, such homogeneity would appear advantageous. However, it is now clear that retinal topographic variations exist across mammalian species in a variety of forms and patterns. We briefly review some of the more established topographic variations in retinas of different mammalian species and focus on the recent discovery that cells belonging to a single neuronal subtype may exhibit distinct topographic variations in distribution, morphology, and even function. We concentrate on the mouse retina-originally viewed as homogenous-in which genetic labeling of distinct neuronal subtypes and other advanced techniques have revealed unexpected anatomical and physiological topographic variations. Notably, different subtypes reveal different patterns of nonuniformity, which may even be opposite or orthogonal to one another. These topographic variations in the encoding of visual space should be considered when studying visual processing in the retina and beyond.

Antagonistic Center-Surround Mechanisms for Direction Selectivity in the Retina.

An antagonistic center-surround receptive field is a key feature in sensory processing, but how it contributes to specific computations such as direction selectivity is often unknown. Retinal On-starburst amacrine cells (SACs), which mediate direction selectivity in direction-selective ganglion cells (DSGCs), exhibit antagonistic receptive field organization: depolarizing to light increments and decrements in their center and surround, respectively. We find that a repetitive stimulation exhausts SAC center and enhances its surround and use it to study how center-surround responses contribute to direction selectivity. Center, but not surround, activation induces direction-selective responses in SACs. Nevertheless, both SAC center and surround elicited direction-selective responses in DSGCs, but to opposite directions. Physiological and modeling data suggest that the opposing direction selectivity can result from inverted temporal balance between excitation and inhibition in DSGCs, implying that SAC's response timing dictates direction selectivity. Our findings reveal antagonistic center-surround mechanisms for direction selectivity and demonstrate how context-dependent receptive field reorganization enables flexible computations.

Low-pass filter properties of basal ganglia cortical muscle loops in the normal and MPTP primate model of parkinsonism.

Oscillatory bursting activity is commonly found in the basal ganglia (BG) and the thalamus of the parkinsonian brain. The frequency of these oscillations is often similar to or higher than that of the parkinsonian tremor, but their relationship to the tremor and other parkinsonian symptoms is still under debate. We studied the frequency dependency of information transmission in the cortex-BG and cortex-periphery loops by recording simultaneously from multiple electrodes located in the arm-related primary motor cortex (MI) and in the globus pallidus (GP) of two vervet monkeys before and after 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treatment and induction of parkinsonian symptoms. We mimicked the parkinsonian bursting oscillations by stimulating with 35 ms bursts given at different frequencies through microelectrodes located in MI or GP while recording the evoked neuronal and motor responses. In the normal state, microstimulation of MI or GP does not modulate the discharge rate in the other structure. However, the functional-connectivity between MI and GP is greatly enhanced after MPTP treatment. In the frequency domain, GP neurons usually responded equally to 1-15 Hz stimulation bursts in both states. In contrast, MI neurons demonstrated low-pass filter properties, with a cutoff frequency above 5 Hz for the MI stimulations, and below 5 Hz for the GP stimulations. Finally, muscle activation evoked by MI microstimulation was markedly attenuated at frequencies higher than 5 Hz. The low-pass properties of the pathways connecting GP to MI to muscles suggest that parkinsonian tremor is not directly driven by the BG 5-10 Hz burst oscillations despite their similar frequencies.

Basal ganglia oscillations and pathophysiology of movement disorders.

Low frequency rest tremor is one of the cardinal signs of Parkinson's disease and some of its animal models. Current physiological studies and models of the basal ganglia differ as to which aspects of neuronal activity are crucial to the pathophysiology of Parkinson's disease. There is evidence that neural oscillations and synchronization play a central role in the generation of the disease. However, parkinsonian tremor is not strictly correlated with the synchronous oscillations in the basal ganglia networks. Rather, abnormal basal ganglia output enforces abnormal thalamo-cortical processing leading to akinesia, the main negative symptom of Parkinson's disease. Parkinsonian tremor has probably evolved as a downstream compensatory mechanism.

Flexible Neural Hardware Supports Dynamic Computations in Retina.

The ability of the retina to adapt to changes in mean light intensity and contrast is well known. Classically, however, adaptation is thought to affect gain but not to change the visual modality encoded by a given type of retinal neuron. Recent findings reveal unexpected dynamic properties in mouse retinal neurons that challenge this view. Specifically, certain cell types change the visual modality they encode with variations in ambient illumination or following repetitive visual stimulation. These discoveries demonstrate that computations performed by retinal circuits with defined architecture can change with visual input. Moreover, they pose a major challenge for central circuits that must decode properties of the dynamic visual signal from retinal outputs.

Inhomogeneous Encoding of the Visual Field in the Mouse Retina.

Stimulus characteristics of the mouse's visual field differ above and below the skyline. Here, we show for the first time that retinal ganglion cells (RGCs), the output neurons of the retina, gradually change their functional properties along the ventral-dorsal axis to allow better representation of the different stimulus characteristics. We conducted two-photon targeted recordings of transient-Offα-RGCs and found that they gradually became more sustained along the ventral-dorsal axis, revealing >5-fold-longer duration responses in the dorsal retina. Using voltage-clamp recordings, pharmacology, and genetic manipulation, we demonstrated that the primary rod pathway underlies this variance. Our findings challenge the current belief that RGCs of the same subtype exhibit the same light responses, regardless of retinal location, and suggest that networks underlying RGC responses may change with retinal location to enable optimized sampling of the visual image.

Dopamine replacement therapy does not restore the full spectrum of normal pallidal activity in the 1-methyl-4-phenyl-1,2,3,6-tetra-hydropyridine primate model of Parkinsonism.

Current physiological studies emphasize the role of neuronal oscillations and synchronization in the pathophysiology of Parkinson's disease; however, little is known about their specific roles in the neuronal substrate of dopamine replacement therapy (DRT). We investigated oscillatory activity and correlations throughout the different states of levodopa-naive parkinsonism as well as "Off-On" and dyskinetic states of DRT in the external globus pallidum (GPe) of tremulous (vervet) and rigid-akinetic (macaque) monkeys and in the internal globus pallidum (GPi) of the vervet monkey. We found that, although oscillatory activity of cells and interneuronal correlation in both pallidal segments increases after induction of parkinsonism with 1-methyl-4-phenyl-1,2,3,6-tetra-hydropyridine (MPTP) and decreases in response to DRT, important differences exist between the two pallidal segments. In the GPi, the fraction of oscillatory cells and relative power of oscillations were significantly higher than in the GPe, and the dominant frequency was within the range of 7.5-13.5 Hz compared with a range of 4.5-7.5 Hz within the GPe. The interneuronal correlations were mostly oscillatory in the GPi, whereas at least half are non-oscillatory in the GPe. We demonstrate that the tremor characteristics after exposure to DRT do not resemble those of the normal or the levodopa-naive state. Moreover, although DRT reverses the MPTP-induced neuronal changes (rate, pattern, and pairwise correlations), the balance between GPe and GPi fails to restore. We therefore suggest that this imbalance reflects additional abnormal organization of the basal ganglia networks in response to dopamine replacement and may constitute the physiological substrate of the limitations and side effects of chronic DRT.

Physiology and pathophysiology of the basal ganglia-thalamo-cortical networks.

Low-frequency resting tremor is one of the cardinal signs of Parkinson's disease (PD) and occurs also in some of its animal models. Current physiological studies and models of the basal ganglia indicate that changes of discharge pattern and synchronization of basal ganglia neurons rather than modification in their discharge rate are crucial to the pathophysiology of PD. However, parkinsonian tremor is not strictly correlated with the synchronous oscillations in the basal ganglia networks. We therefore suggest that abnormal basal ganglia output enforces abnormal thalamo-cortical processing leading to akinesia, the main negative symptom of Parkinson's disease. The parkinsonian positive motor signs, such as tremor and rigidity, most likely evolve as a downstream compensatory mechanism.

Visual stimulation switches the polarity of excitatory input to starburst amacrine cells.

Direction-selective ganglion cells (DSGCs) are tuned to motion in one direction. Starburst amacrine cells (SACs) are thought to mediate this direction selectivity through precise anatomical wiring to DSGCs. Nevertheless, we previously found that visual adaptation can reverse DSGCs's directional tuning, overcoming the circuit anatomy. Here we explore the role of SACs in the generation and adaptation of direction selectivity. First, using pharmacogenetics and two-photon calcium imaging, we validate that SACs are necessary for direction selectivity. Next, we demonstrate that exposure to an adaptive stimulus dramatically alters SACs' synaptic inputs. Specifically, after visual adaptation, On-SACs lose their excitatory input during light onset but gain an excitatory input during light offset. Our data suggest that visual stimulation alters the interactions between rod- and cone-mediated inputs that converge on the terminals of On-cone BCs. These results demonstrate how the sensory environment can modify computations performed by anatomically defined neuronal circuits.

On and off retinal circuit assembly by divergent molecular mechanisms.

Direction-selective responses to motion can be to the onset (On) or cessation (Off) of illumination. Here, we show that the transmembrane protein semaphorin 6A and its receptor plexin A2 are critical for achieving radially symmetric arborization of On starburst amacrine cell (SAC) dendrites and normal SAC stratification in the mouse retina. Plexin A2 is expressed in both On and Off SACs; however, semaphorin 6A is expressed in On SACs. Specific On-Off bistratified direction-selective ganglion cells in semaphorin 6A(-/-) mutants exhibit decreased tuning of On directional motion responses. These results correlate the elaboration of symmetric SAC dendritic morphology and asymmetric responses to motion, shedding light on the development of visual pathways that use the same cell types for divergent outputs.

Visual stimulation reverses the directional preference of direction-selective retinal ganglion cells.

Direction selectivity in the retina is mediated by direction-selective ganglion cells. These cells are part of a circuit in which they are asymmetrically wired to inhibitory neurons. Thus, they respond strongly to an image moving in the preferred direction and weakly to an image moving in the opposite (null) direction. Here, we demonstrate that adaptation with short visual stimulation of a direction-selective ganglion cell using drifting gratings can reverse this cell's directional preference by 180°. This reversal is robust, long lasting, and independent of the animal's age. Our findings indicate that, even within circuits that are hardwired, the computation of direction can be altered by dynamic circuit mechanisms that are guided by visual stimulation.

Closed-loop deep brain stimulation is superior in ameliorating parkinsonism.

Continuous high-frequency deep brain stimulation (DBS) is a widely used therapy for advanced Parkinson's disease (PD) management. However, the mechanisms underlying DBS effects remain enigmatic and are the subject of an ongoing debate. Here, we present and test a closed-loop stimulation strategy for PD in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) primate model of PD. Application of pallidal closed-loop stimulation leads to dissociation between changes in basal ganglia (BG) discharge rates and patterns, providing insights into PD pathophysiology. Furthermore, cortico-pallidal closed-loop stimulation has a significantly greater effect on akinesia and on cortical and pallidal discharge patterns than standard open-loop DBS and matched control stimulation paradigms. Thus, closed-loop DBS paradigms, by modulating pathological oscillatory activity rather than the discharge rate of the BG-cortical networks, may afford more effective management of advanced PD. Such strategies have the potential to be effective in additional brain disorders in which a pathological neuronal discharge pattern can be recognized.

Computational physiology of the basal ganglia in Parkinson's disease.

The normal activity of basal ganglia neurons is characterized by Poisson-like (random) firing patterns. Correlations between neurons of the same structure are weak or non-existent. By contrast, synchronous oscillations are commonly found in the basal ganglia of human patients and animal models of Parkinson's disease. The frequency of these oscillations is often similar to that of the parkinsonian tremor, but their role in generating the tremor or other parkinsonian symptoms is still under debate. The tremor is intermittent and does not appear in all human patients. Similarly, primate models tend to develop tremor as a function of species of monkey. African green (vervet) monkeys usually demonstrate a high-amplitude, low-frequency (4-7Hz) tremor beyond their akinesia and bradykinesia, whereas macaques tend to be akinetic rigid and rarely demonstrate a low-amplitude high-frequency (10-12Hz) action-postural tremor. We took advantage of this fact and studied the appearance of the synchronicity and oscillations in six monkeys, three vervets and three macaques, before and after 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) systemic treatment and induction of parkinsonism. Multiple extracellular recordings were conducted in the primary motor cortex of two monkeys and in the globus pallidus (GP) of all six monkeys. All the monkeys became akinetic and bradykinetic as a result of the MPTP treatment, but only vervets demonstrated prolonged episodes of low-frequency (4-6Hz) tremor, whereas macaques were non-tremulous. The GP population exhibited approximately 5Hz oscillatory activity in all six monkeys, whereas approximately 10Hz neural oscillations were only detected in the tremulous monkeys. The activity of the cortical neurons became strongly oscillatory at approximately 10Hz in one of these monkeys, but not the other, although both were tremulous and exhibited comparable pallidal oscillatory activity. Finally, synchronous oscillations, when present, were centred around the higher frequencies of oscillations. These findings suggest that there is a correlation between high-frequency GP neural oscillations and tremor. Furthermore, these pallidal 10Hz oscillations are probably transferred to the periphery through cortical and brainstem pathways.

Local shuffling of spike trains boosts the accuracy of spike train spectral analysis.

Spectral analysis of neuronal spike trains is an important tool in understanding the characteristics of neuronal activity by providing insights into normal and pathological periodic oscillatory phenomena. However, the refractory period creates high-frequency modulations in spike-train firing rate because any rise in the discharge rate causes a descent in subsequent time bins, leading to multifaceted modifications in the structure of the spectrum. Thus the power spectrum of the spiking activity (autospectrum) displays elevated energy in high frequencies relative to the lower frequencies. The spectral distortion is more dominant in neurons with high firing rates and long refractory periods and can lead to reduced identification of low-frequency oscillations (such as the 5- to 10-Hz burst oscillations typical of Parkinsonian basal ganglia and thalamus). We propose a compensation process that uses shuffling of interspike intervals (ISIs) for reliable identification of oscillations in the entire frequency range. This compensation is further improved by local shuffling, which preserves the slow changes in the discharge rate that may be lost in global shuffling. Cross-spectra of pairs of neurons are similarly distorted regardless of their correlation level. Consequently, identification of low-frequency synchronous oscillations, even for two neurons recorded by a single electrode, is improved by ISI shuffling. The ISI local shuffling is computed with confidence limits that are based on the first-order statistics of the spike trains, thus providing a reliable estimation of auto- and cross-spectra of spike trains and making it an optimal tool for physiological studies of oscillatory neuronal phenomena.