Layer 4C Research Articles

Comparison of orientation encoding across layers within single columns of primate V1 revealed by high-density recordings.

Primary visual cortex (V1) has been the focus of extensive neurophysiological investigations, with its laminar organization serving as a crucial model for understanding the functional logic of neocortical microcircuits. Utilizing newly developed high-density, Neuropixels probes, we measured visual responses from large populations of simultaneously recorded neurons distributed across layers of macaque V1. Within single recordings, myriad differences in the functional properties of neuronal subpopulations could be observed. Notably, while standard measurements of orientation selectivity showed only minor differences between laminar compartments, decoding stimulus orientation from layer 4C responses outperformed both superficial and deep layers within the same cortical column. The superior orientation discrimination within layer 4C was associated with greater response reliability of individual neurons rather than lower correlated activity within neuronal populations. Our results underscore the efficacy of high-density electrophysiology in revealing the functional organization and network properties of neocortical microcircuits within single experiments.

Explaining the formation of the 36 smallest known atomic isotopes: from hydrogen to krypton

This work presents a novel approach to understanding the formation and structure of the 36 smallest known atomic isotopes, ranging from hydrogen to krypton, through the lens of the KUPT model. Focusing on the Small Known Isotope (SKI) of each atom, this study meticulously examines the distribution of protons across five nuclear layers (1E, 2T, 3T, 4E, and 4C), constructing a proton tree that spans from hydrogen to krypton atoms. With the aid of Artificial Intelligence, specifically Chat GPT-4, an in-depth analysis was conducted for each atom within the KUPT model framework. This process involved defining minimum stability rules for the nuclear structure, particularly regarding neutron inclusion, across the model’s first five layers. Remarkably, in 35 out of 36 cases, the KUPT model’s predictions aligned perfectly with the SKI data, underscoring the model’s accuracy and reliability. However, an intriguing discrepancy emerged in the case of the Gallium atom (62Ga), suggesting the possible existence of an unknown, lighter isotope with two or three fewer protons (59Ga or 60Ga). This discrepancy not only highlights the KUPT model’s predictive capability but also points to a fertile ground for further experimental investigation, aiming to uncover potentially undiscovered isotope forms of Gallium.

Presented to Kepler Ulianov Proton Tree, A Way to Distribute Protons and Neutrons Within the Atomic Nucleus. First part: From Hydrogen to Krypton

This work presents a novel approach to understanding the formation and structure of the 36 smallest known atomic isotopes, ranging from hydrogen to krypton, through the lens of the KUPT model. Focusing on the Small Known Isotope (SKI) of each atom, this study meticulously examines the distribution of protons across five nuclear layers (1E, 2T, 3T, 4E, and 4C), constructing a proton tree that spans from hydrogen to krypton atoms. With the aid of Artificial Intelligence, specifically Chat GPT-4, an in-depth analysis was conducted for each atom within the KUPT model framework. This process involved defining minimum stability rules for the nuclear structure, particularly regarding neutron inclusion, across the model’s first five layers. Remarkably, in 35 out of 36 cases, the KUPT model’s predictions aligned perfectly with the SKI data, underscoring the model’s accuracy and reliability. However, an intriguing discrepancy emerged in the case of the Gallium atom (62Ga), suggesting the possible existence of an unknown, lighter isotope with two or three fewer protons (59Ga or 60Ga). This discrepancy not only highlights the KUPT model’s predictive capability but also points to a fertile ground for further experimental investigation, aiming to uncover potentially undiscovered isotope forms of Gallium.

Balkan Neanderthals: The Late Pleistocene palaeoecological sequence of Pešturina Cave (Niš, Serbia)

The Central Balkans are a key biogeographical region in Southern Europe, influenced by a central European-Mediterranean climate, which acted as a refugium for flora and fauna, and favored the dispersion of Neanderthals and migration of modern human populations during Late Glacial Period. This study presents pollen analyses of sediment and hyaena coprolites from Pešturina Cave in Serbia to reconstruct the vegetation landscapes faced by Balkan Neanderthals and early Anatomically Modern Humans between MIS 5e-3. Between MIS 5e-5c (archaeological layers 4c and 4b) and MIS 5b-5a (layer 4a), semi-forested environments prevailed, characterized by Pinus, deciduous Quercus, Tilia and other angiosperm woody taxa, accompanied by heliophytes such as Artemisia and Poaceae. During MIS 4-3 (layers 3-2), the vegetation was dominated by Artemisia-Poaceae steppes with Quercus patches, conifers and legumes. Overall across the sequence, pollen assemblages are highly diverse and include a number of deciduous trees and sclerophylls. In addition, the occurrence of several herbaceous taxa reinforces the view that the Balkans were outstanding for endemicity. Neanderthals and early Upper Palaeolithic hominins lived in a highly diverse refugium, offering multiple opportunities for survival during the warm interstadials and, more critically, the cold stadials of the Pleistocene.

A laminar model for the joint development of ocular dominance columns and CO blobs in the primary visual cortex

In this paper, we present a multi-layer, activity-dependent model for the joint development of ocular dominance (OD) columns and cytochrome oxidase (CO) blobs in primate V1. For simplicity, we focus on layers 4C and 2/3 with both layers receiving direct thalamic inputs and layer 4C sending vertical projections to layer 2/3. Both the thalamic and the vertical connections are taken to be modifiable by activity. Using a correlation-based Hebbian learning rule with subtractive normalization, we show how the formation of an OD map in layer 4C is inherited by layer 2/3 via the vertical projections. Competition between these feedforward projections and the direct thalamic input to layer 2/3 then results in the formation of CO blobs superimposed upon the ocular dominance map. The spacing of the OD columns is determined by the spatial profile of the intralaminar connections within layer 4, while the spacing of CO blobs depends both on the width of the OD columns inherited from layer 4 and the spatial distribution of intralaminar connections within the superficial layer. The resulting CO blob distribution is shown to be consistent with experimental data. In addition, we numerically simulate monocular deprivation and find that while the CO blob distribution is unaltered, the OD pattern undergoes modification. The OD stripes of the deprived eye narrow, whereas the OD stripes for the remaining open eye widen.

Mechanism underpinning the sharpening of orientation and spatial frequency selectivities in the tree shrew (Tupaia belangeri) primary visual cortex.

Most neurons in the primary visual cortex (V1) of mammals show sharp orientation selectivity and band-pass spatial frequency tuning. Here, we examine whether sharpening of the broad tuning that exists subcortically, namely in the retina and the lateral geniculate nucleus (LGN), underlie the sharper tuning seen for both the above features in tree shrew V1. Since the transition from poor feature selectivity to sharp tuning occurs entirely within V1 in tree shrews, we examined the orientation selectivity and spatial frequency tuning of neurons within individual electrode penetrations. We found that most layer 4 and layer 2/3 neurons in the same cortical column preferred the same stimulus orientation. However, a subset of layer 3c neurons close to the layer 4 border preferred near orthogonal orientations, suggesting that layer 2/3 neurons may inherit the orientation preferences of their layer 4 input neurons and also receive cross-orientation inhibition from layer 3c neurons. We also found that layer 4 neurons showed sharper orientation selectivity at higher spatial frequencies, suggesting that attenuation of low spatial frequency responses by spatially broad inhibition acting on layer 4 inputs to layer 2/3 neurons can enhance both orientation and spatial frequency selectivities. However, in a proportion of layer 2/3 neurons, the sharper tuning of layer 2/3 neurons appeared to arise also or even mainly from inhibition specific to high spatial frequencies acting on the layer 4 inputs to layer 2/3. Overall, our results are consistent with the suggestion that in tree shrews, sharp feature selectivity in layer 2/3 can be established by intracortical mechanisms that sharpen biases observed in layer 4, which are in turn inherited presumably from thalamic afferents.

Cubic Silicon Carbide (3C–SiC) as a buffer layer for high efficiency and highly stable CdTe solar cell

Cubic Silicon Carbide (3C–SiC) is a potential material for use in photovoltaics for its significant advancement in growth in terms of crystal quality and domain size. Hence, 3C–SiC has been introduced here as an alternative non-toxic buffer layer for heterojunction CdTe solar cell. The solar cell is designed for the study consisted of multi-junction semiconductor layers such as p-CdTe/n-3C–SiC/n-SnO2. Device modeling of the novel CdTe solar cell including the thickness and doping concentration of the novel buffer layer 3C–SiC are investigated and optimized using SCAPS-1D software. Also, the defect density in the buffer layer is studied to see the tolerance of the proposed device structure. The working temperature and incident light intensity are also varied to deduce the effect of environmental conditions on efficient PV operation.

Diversity amongst human cortical pyramidal neurons revealed via their sag currents and frequency preferences

In the human neocortex coherent interlaminar theta oscillations are driven by deep cortical layers, suggesting neurons in these layers exhibit distinct electrophysiological properties. To characterize this potential distinctiveness, we use in vitro whole-cell recordings from cortical layers 2 and 3 (L2&3), layer 3c (L3c) and layer 5 (L5) of the human cortex. Across all layers we observe notable heterogeneity, indicating human cortical pyramidal neurons are an electrophysiologically diverse population. L5 pyramidal cells are the most excitable of these neurons and exhibit the most prominent sag current (abolished by blockade of the hyperpolarization activated cation current, Ih). While subthreshold resonance is more common in L3c and L5, we rarely observe this resonance at frequencies greater than 2 Hz. However, the frequency dependent gain of L5 neurons reveals they are most adept at tracking both delta and theta frequency inputs, a unique feature that may indirectly be important for the generation of cortical theta oscillations.

Laminar Subnetworks of Response Suppression in Macaque Primary Visual Cortex.

Cortical inhibition plays an important role in information processing in the brain. However, the mechanisms by which inhibition and excitation are coordinated to generate functions in the six layers of the cortex remain unclear. Here, we measured laminar-specific responses to stimulus orientations in primary visual cortex (V1) of awake monkeys (male, Macaca mulatta). We distinguished inhibitory effects (suppression) from excitation, by taking advantage of the separability of excitation and inhibition in the orientation and time domains. We found two distinct types of suppression governing different layers. Fast suppression (FS) was strongest in input layers (4C and 6), and slow suppression (SS) was 3 times stronger in output layers (2/3 and 5). Interestingly, the two types of suppression were correlated with different functional properties measured with drifting gratings. FS was primarily correlated with orientation selectivity in input layers (r = −0.65, p < 10−9), whereas SS was primarily correlated with surround suppression in output layers (r = 0.61, p < 10−4). The earliest SS in layer 1 indicates the origin of cortical feedback for SS, in contrast to the feedforward/recurrent origin of FS. Our results reveal two V1 laminar subnetworks with different response suppression that may provide a general framework for laminar processing in other sensory cortices.SIGNIFICANCE STATEMENT This study sought to understand inhibitory effects (suppression) and their relationships with functional properties in the six different layers of the cortex. We found that the diversity of neural responses across layers in primary visual cortex (V1) could be fully explained by one excitatory and two suppressive components (fast and slow suppression). The distinct laminar distributions, origins, and functional roles of the two types of suppression provided a simplified representation of the differences between two V1 subnetworks (input network and output network). These results not only help to elucidate computational principles in macaque V1, but also provide a framework for general computation of cortical laminae in other sensory cortices.

GABAergic and non-GABAergic subpopulations of Kv3.1b-expressing neurons in macaque V2 and MT: laminar distributions and proportion of total neuronal population.

The Kv3.1b potassium channel subunit, which facilitates the fast-spiking phenotype characteristic of parvalbumin (PV)-expressing inhibitory interneurons, is also expressed by subpopulations of excitatory neurons in macaque cortex. We have previously shown that V1 neurons expressing Kv3.1b but not PV or GABA were largely concentrated within layers 4Cα and 4B of V1, suggesting laminar or pathway specificity. In the current study, the distribution and pattern of co-immunoreactivity of GABA, PV, and Kv3.1b across layers in extrastriate cortical areas V2 and MT of the macaque monkey were measured using the same triple immunofluorescence labeling, confocal microscopy, and partially automated cell-counting strategies used in V1. For comparison, densities of the overall cell and neuronal populations were also measured for each layer of V2 and MT using tissue sections immunofluorescence labeled for the pan-neuronal marker NeuN. GABAergic neurons accounted for 14% of the total neuronal population in V2 and 25% in MT. Neurons expressing Kv3.1b but neither GABA nor PV were present in both areas. This subpopulation was most prevalent in the lowest subcompartment of layer 3, comprising 5% of the total neuronal population in layer 3C of both areas, and 41% and 36% of all Kv3.1b+ neurons in this layer in V2 and MT, respectively. The prevalence and laminar distribution of this subpopulation were remarkably consistent between V2 and MT and showed a striking similarity to the patterns observed previously in V1, suggesting a common contribution to the cortical circuit across areas.

Cholinergic Modulation Promotes Attentional Modulation in Primary Visual Cortex- A Modeling Study

Attention greatly influences sensory neural processing by enhancing firing rates of neurons that represent the attended stimuli and by modulating their tuning properties. The cholinergic system is believed to partly mediate the attention contingent improvement of cortical processing by influencing neuronal excitability, synaptic transmission and neural network characteristics. Here, we used a biophysically based model to investigate the mechanisms by which cholinergic system influences sensory information processing in the primary visual cortex (V1) layer 4C. The physiological properties and architectures of our model were inspired by experimental data and include feed-forward input from dorsal lateral geniculate nucleus that sets up orientation preference in V1 neural responses. When including a cholinergic drive, we found significant sharpening in orientation selectivity, desynchronization of LFP gamma power and spike-field coherence, decreased response variability and correlation reduction mostly by influencing intracortical interactions and by increasing inhibitory drive. Our results indicated that these effects emerged due to changes specific to the behavior of the inhibitory neurons. The behavior of our model closely resembles the effects of attention on neural activities in monkey V1. Our model suggests precise mechanisms through which cholinergic modulation may mediate the effects of attention in the visual cortex.

A Neuronal Network Model of the Primate Visual System: Color Mechanisms in the Retina, LGN and V1.

Color plays a key role in human vision but the neural machinery that underlies the transformation from stimulus to perception is not well understood. Here, we implemented a two-dimensional network model of the first stages in the primate parvocellular pathway (retina, lateral geniculate nucleus and layer 4C in V1) consisting of conductance-based point neurons. Model parameters were tuned based on physiological and anatomical data from the primate foveal and parafoveal vision, the most relevant visual field areas for color vision. We exhaustively benchmarked the model against well-established chromatic and achromatic visual stimuli, showing spatial and temporal responses of the model to disk- and ring-shaped light flashes, spatially uniform squares and sine-wave gratings of varying spatial frequency. The spatiotemporal patterns of parvocellular cells and cortical cells are consistent with their classification into chromatically single-opponent and double-opponent groups, and nonopponent cells selective for luminance stimuli. The model was implemented in the widely used neural simulation tool NEST and released as open source software. The aim of our modeling is to provide a biologically realistic framework within which a broad range of neuronal interactions can be examined at several different levels, with a focus on understanding how color information is processed.

Major Feedforward Thalamic Input Into Layer 4C of Primary Visual Cortex in Primate.

One of the underlying principles of how mammalian circuits are constructed is the relative influence of feedforward to recurrent synaptic drive. It has been dogma in sensory systems that the thalamic feedforward input is relatively weak and that there is a large amplification of the input signal by recurrent feedback. Here we show that in trichromatic primates there is a major feedforward input to layer 4C of primary visual cortex. Using a combination of 3D-electron-microscopy and 3D-confocal imaging of thalamic boutons we found that the average feedforward contribution was about 20% of the total excitatory input in the parvocellular (P) pathway, about 3 times the currently accepted values for primates. In the magnocellular (M) pathway it was around 15%, nearly twice the currently accepted values. New methods showed the total synaptic and cell densities were as much as 150% of currently accepted values. The new estimates of contributions of feedforward synaptic inputs into visual cortex call for a major revision of the design of the canonical cortical circuit.

Contrasting effects of strabismic amblyopia on metabolic activity in superficial and deep layers of striate cortex.

To probe the mechanism of visual suppression, we have raised macaques with strabismus by disinserting the medial rectus muscle in each eye at 1 mo of age. Typically, this operation produces a comitant, alternating exotropia with normal acuity in each eye. Here we describe an unusual occurrence: the development of severe amblyopia in one eye of a monkey after induction of exotropia. Shortly after surgery, the animal demonstrated a strong fixation preference for the left eye, with apparent suppression of the right eye. Later, behavioral testing showed inability to track or to saccade to targets with the right eye. With the left eye occluded, the animal demonstrated no visually guided behavior. Optokinetic nystagmus was absent in the right eye. Metabolic activity in striate cortex was assessed by processing the tissue for cytochrome oxidase (CO). Amblyopia caused loss of CO in one eye's rows of patches, presumably those serving the blind eye. Layers 4A and 4B showed columns of reduced CO, in register with pale rows of patches in layer 2/3. Layers 4C, 5, and 6 also showed columns of CO activity, but remarkably, comparison with more superficial layers showed a reversal in contrast. In other words, pale CO staining in layers 2/3, 4A, and 4B was aligned with dark CO staining in layers 4C, 5, and 6. No experimental intervention or deprivation paradigm has been reported previously to produce opposite effects on metabolic activity in layers 2/3, 4A, and 4B vs. layers 4C, 5, and 6 within a given eye's columns.

Neuroanatomical distribution of oxytocin and vasopressin 1a receptors in the socially monogamous coppery titi monkey (Callicebus cupreus)

The coppery titi monkey (Callicebus cupreus) is a socially monogamous New World primate that has been studied in the field and the laboratory to investigate the behavioral neuroendocrinology of primate pair bonding and parental care. Arginine vasopressin has been shown to influence male titi monkey pair-bonding behavior, and studies are currently underway to examine the effects of oxytocin on titi monkey behavior and physiology. Here, we use receptor autoradiography to identify the distribution of arginine vasopressin 1a receptor (AVPR1a) and oxytocin receptors (OXTR) in hemispheres of titi monkey brain (n=5). AVPR1a are diffuse and widespread throughout the brain, but the OXTR distribution is much more limited, with the densest binding being in the hippocampal formation (dentate gyrus, CA1 field) and the presubiculum (layers I and III). Moderate OXTR binding was detected in the nucleus basalis of Meynert, pulvinar, superior colliculus, layer 4C of primary visual cortex, periaqueductal gray (PAG), pontine gray, nucleus prepositus, and spinal trigeminal nucleus. OXTR mRNA overlapped with OXTR radioligand binding, confirming that the radioligand was detecting OXTR protein. AVPR1a binding is present throughout the cortex, especially in cingulate, insular, and occipital cortices, as well as in the caudate, putamen, nucleus accumbens, central amygdala, endopiriform nucleus, hippocampus (CA4 field), globus pallidus, lateral geniculate nucleus, infundibulum, habenula, PAG, substantia nigra, olivary nucleus, hypoglossal nucleus, and cerebellum. Furthermore, we show that, in the titi monkey brain, the OXTR antagonist ALS-II-69 is highly selective for OXTR and that the AVPR1a antagonist SR49059 is highly selective for AVPR1a. Based on these results and the fact that both ALS-II-69 and SR49059 are non-peptide, small-molecule antagonists that should be capable of crossing the blood–brain barrier, these two compounds emerge as excellent candidates for the pharmacological manipulation of OXTR and AVPR1a in future behavioral experiments in titi monkeys and other primate species.

Distribution of vesicular glutamate transporter 2 (VGluT2) in the primary visual cortex of the macaque and human

The majority of thalamic terminals in V1 arise from lateral geniculate nucleus (LGN) afferents. Thalamic afferent terminals are preferentially labeled by an isoform of the vesicular glutamate transporter, VGluT2. The goal of our study was to determine the distribution of VGluT2-ir puncta in macaque and human visual cortex. First, we investigated the distribution of VGluT2-ir puncta in all layers of macaque monkey primary visual cortex (V1), and found a very close correspondence between the known distribution of LGN afferents from previous studies and the distribution of VGluT2-immunoreactive (-ir) puncta. There was also a close correspondence between cytochrome oxidase density and VGluT2-ir puncta distribution. After validating the correspondence in macaque, we made a comparative study in human V1. In many aspects, the distribution of VGluT2-ir puncta in human was qualitatively similar to that of the macaque: high densities in layer 4C, patches of VGluT2-ir puncta in the supragranular layer (2/3), lower but clear distribution in layers 1 and 6, and very few puncta in layers 5 and 4B. However, there were also important differences between macaques and humans. In layer 4A of human, there was a sparse distribution of VGluT2-ir puncta, whereas in macaque, there was a dense distribution with the characteristic honeycomb organization. The results suggest important changes in the pattern of cortical VGluT2 immunostaining that may be related to evolutionary differences in the cortical organization of LGN afferents between Old World monkeys and humans.

The Contributions of the on and off Gain Difference to the Contextual Effect in Macaque Monkey V1

In macaque monkey V1, the response amplitudes of ON and OFF sub-regions of simple receptive fields are about equal for cells in the input layer 4c, but the OFF responses tend to be greater than the ON responses for layer-2/3 cells. This OFF-over-ON bias is evident when receptive fields are mapped with sparse noise (a series of dark or bright squares shown in different space and time) but is weaker with Hartleys (gratings shown briefly at different orientations and spatial frequencies). One possible mechanism for the receptive-field mismatch in layer 2/3 of V1 is the difference between ON and OFF response gains caused by different degrees of sparseness between Hartleys and sparse noise. Here we manipulated the relative strength of ON and OFF sparse-noise responses until the receptive field similarity (RFS, quantified as the pixel-by-pixel correlation between two maps) between sparse-noise and Hartley maps reached the maximum. On average, the relative strength of ON/OFF responses in sparse-noise maps should be increased to approximately two times of their origins in order to match closely their Hartley compartments for layer-2/3 neurons (1.93±0.19; mean±S.E.M.). The sole change in response gain largely increased the RFS between Hartley and sparse-noise maps (on average the RFS increases from 0.35 to 0.58), and it could account for approximately 81% of the overall differences between the two maps. In summary, the difference in ON and OFF gains under different visual stimulations is one critical mechanism underlying the contextual effect in the superficial layer of V1.

Cholinergic suppression of visual responses in primate V1 is mediated by GABAergic inhibition

Acetylcholine (ACh) has been implicated in selective attention. To understand the local circuit action of ACh, we iontophoresed cholinergic agonists into the primate primary visual cortex (V1) while presenting optimal visual stimuli. Consistent with our previous anatomical studies showing that GABAergic neurons in V1 express ACh receptors to a greater extent than do excitatory neurons, we observed suppressed visual responses in 36% of recorded neurons outside V1's primary thalamorecipient layer (4c). This suppression is blocked by the GABA(A) receptor antagonist gabazine. Within layer 4c, ACh release produces a response gain enhancement (Disney AA, Aoki C, Hawken MJ. Neuron 56: 701-713, 2007); elsewhere, ACh suppresses response gain by strengthening inhibition. Our finding contrasts with the observation that the dominant mechanism of suppression in the neocortex of rats is reduced glutamate release. We propose that in primates, distinct cholinergic receptor subtypes are recruited on specific cell types and in specific lamina to yield opposing modulatory effects that together increase neurons' responsiveness to optimal stimuli without changing tuning width.

Neuronal Projections from V1 to V2 in Amblyopia

The mechanism of amblyopia in children with congenital cataract is not understood fully, but studies in macaques have shown that geniculate synapses are lost in striate cortex (V1). To search for other projection abnormalities in amblyopia, the pathway from V1 to V2 was examined using a triple-label technique in three animals raised with monocular suture. [(3)H]proline was injected into one eye to label the ocular dominance columns. Cholera toxin B subunit conjugated to gold (CTB-Au) was injected into V2 to label V1 projection neurons. Alternate sections were processed for cytochrome oxidase (CO) and CTB-Au, or dipped for autoradiography. Eight fields of CTB-Au-labeled cells in V1 opposite injection sites were plotted in layers 2/3 or 4B. After thin stripe injection, labeled cells were concentrated in CO patches. Despite column shrinkage, cells in deprived and normal columns were equal in size and density in both layers 2/3 and 4B. After pale or thick stripe injection, labeled cells were concentrated in interpatches. Only 23% of projection neurons originated from deprived columns. This reduction exceeded the degree of column shrinkage, a result explained by the fact that column shrinkage causes disproportionate loss of interpatch territory. These data indicate that early monocular form deprivation does not alter the segregation of patch and interpatch pathways to V2 stripes or cause selective loss or atrophy of V1 projection neurons. The effect of shrinkage of geniculocortical afferents in layer 4C following visual deprivation is not amplified further by attenuation of the amblyopic eye's projections from V1 to V2.

Cholinergic modulation of response gain in the primary visual cortex of the macaque

ACh modulates neuronal activity throughout the cerebral cortex, including the primary visual cortex (V1). However, a number of issues regarding this modulation remain unknown, such as the effect and its function and the receptor subtypes involved. To address these issues, we combined extracellular single-unit recordings and microiontophoretic administration of ACh and measured V1 neuronal responses to drifting sinusoidal grating stimuli in anesthetized macaque monkeys. ACh was found to have mostly facilitatory effects on the visual responses, although some cases of suppressive effects were also seen. To assess the functional role of ACh, we further examined how ACh modulates the stimulus contrast-response function, finding that the response gain increased with the facilitatory effect. The response facilitation was completely or strongly blocked by atropine (At), a muscarinic ACh receptor (mAChR) antagonist, in almost all neurons (96% of cells), whereas any residual effect after At administration was fully removed by mecamylamine, a nicotinic AChR (nAChR) antagonist, suggesting a predominant role for mAChRs in this mechanism. Furthermore, we found no laminar distribution bias for the facilitatory modulation, although the relative contribution of mAChRs was smaller in layer 4C than in other layers. The suppressive effect was blocked completely by At. These results demonstrate that ACh plays an important role in visual information processing in V1 by controlling the response gain via mAChRs across all cortical layers and via nAChRs, mainly in layer 4C.