Neurons In Ferret Visual Cortex Research Articles

Differential tuning of excitation and inhibition shapes direction selectivity in ferret visual cortex

To encode specific sensory inputs, cortical neurons must generate selective responses for distinct stimulus features. In principle, a variety of factors contribute to a cortical neuron’s response selectivity: the tuning and strength of excitatory1–3 and inhibitory synaptic inputs4–6, dendritic nonlinearities7–9, and spike threshold10,11. Here we employ a combination of techniques including in vivo whole-cell recording, synaptic and cellular resolution in vivo two photon calcium imaging, and GABAergic-selective optogenetic manipulation to dissect the factors contributing to direction selective responses of layer 2/3 neurons in ferret visual cortex (V1). Two-photon calcium imaging of dendritic spines12,13 revealed that each neuron receives a mixture of excitatory synaptic inputs selective for the somatic preferred or null direction of motion. The relative number of preferred- and null-tuned excitatory inputs predicted a neuron’s somatic direction preference, but failed to account for the degree of direction selectivity. In contrast, in vivo whole-cell patch clamp recordings revealed a striking degree of direction selectivity in subthreshold responses that was significantly correlated with spiking direction selectivity. Subthreshold direction selectivity was predicted by the magnitude and variance of the response to the null direction of motion, and several lines of evidence including conductance measurements demonstrate that differential tuning of excitation and inhibition suppresses responses to the null direction of motion. Consistent with this idea, optogenetic inactivation of GABAergic neurons in layer 2/3 reduced direction selectivity by enhancing responses to the null direction. Furthermore, using a new technique to optogenetically map connections of inhibitory neurons in layer 2/3 in vivo, we find that layer 2/3 inhibitory neurons make long-range, intercolumnar projections to excitatory neurons that prefer the opposite direction of motion. We conclude that intracortical inhibition exerts a major influence on the degree of direction selectivity in layer 2/3 of ferret V1 by suppressing responses to the null direction of motion.

GABAergic Neurons in Ferret Visual Cortex Participate in Functionally Specific Networks

Functional circuits in the visual cortex require the coordinated activity of excitatory and inhibitory neurons. Molecular genetic approaches in the mouse have led to the "local non-specific pooling principle" of inhibitory connectivity, in which inhibitory neurons are untuned for stimulus features due to the random pooling of local inputs. However, it remains unclear whether this principle generalizes to species with a columnar organization of feature selectivity such as carnivores, primates, and humans. Here we use virally mediated GABAergic-specific GCaMP6f expression to demonstrate that inhibitory neurons in ferret visual cortex respond robustly and selectively to oriented stimuli. We find that the tuning of inhibitory neurons is inconsistent with the local non-specific pooling of excitatory inputs and that inhibitory neurons exhibit orientation-specific noise correlations with local and distant excitatory neurons. These findings challenge the generality of the non-specific pooling principle for inhibitory neurons, suggesting different rules for functional excitatory-inhibitory interactions in non-murine species.

Influence of contrast on orientation and temporal frequency tuning in ferret primary visual cortex.

Neurons in primary visual cortex are highly sensitive to the contrast, orientation, and temporal frequency of a visual stimulus. These three stimulus properties can be varied independently of one another, raising the question of how they interact to influence neuronal responses. We recorded from individual neurons in ferret primary visual cortex to determine the influence of stimulus contrast on orientation tuning, temporal-frequency tuning, and latency to visual response. Results show that orientation-tuning bandwidth is not affected by contrast level. Thus neurons in ferret visual cortex display contrast-invariant orientation tuning. Stimulus contrast does, however, influence the structure of orientation-tuning curves as measures of circular variance vary inversely with contrast for both simple and complex cells. This change in circular variance depends, in part, on a contrast-dependent change in the ratio of null to preferred orientation responses. Stimulus contrast also has an influence on the temporal-frequency tuning of cortical neurons. Both simple and complex cells display a contrast-dependent rightward shift in their temporal frequency-tuning curves that results in an increase in the highest temporal frequency needed to produce a half-maximum response (TF(50)). Results show that the degree of the contrast-dependent increase in TF(50) is similar for cortical neurons and neurons in the lateral geniculate nucleus (LGN) and indicate that subcortical mechanisms likely play a major role in establishing the degree of effect displayed by downstream neurons. Finally, results show that LGN and cortical neurons experience a contrast-dependent phase advance in their visual response. This phase advance is most pronounced for cortical neurons indicating a role for both subcortical and cortical mechanisms.

Reorganization of Exuberant Axonal Arbors Contributes to the Development of Laminar Specificity in Ferret Visual Cortex

Layer-specific cortical axons are believed to develop precisely from the outset without making exuberant branches in incorrect cortical layers. We tested this hypothesis by following the development of axonal arbors of layer 2/3 pyramidal neurons in ferret visual cortex, which in the adult have dense axonal arbors in layers 2/3 and 5 and not in layer 4. We began our studies at earlier ages and sampled at closer intervals than in previous studies. Our studies reveal that between postnatal day (P) 14 and P18 the initial growth of axonal arbors lacks specificity for layers 2/3 and 5 and involves the formation of a small number of branches incorrectly in layer 4. After P18 the number of incorrect axonal branches in layer 4 decreased, whereas there was extensive axonal growth specifically in layers 2/3 and 5. To further study the mechanisms involved in directing the layer-specific growth and elimination of axonal branches, we studied the development of layer 2/3 neurons in slices kept in culture for 5 d. In these studies the initial imprecise growth at P14–18 followed by precise growth after P18 was mimicked <i>in vitro</i>, but the elimination of incorrect axons from layer 4 did not occur. These studies reveal that most axonal arbors grow precisely in the correct layers, but the initial growth involves the formation of a small number of incorrect axonal branches that are later eliminated. Therefore the mechanisms that regulate axonal growth are not as precise initially as later, and mechanisms must exist to allow selective elimination of axon collaterals from incorrect layers.

Reorganization of exuberant axonal arbors contributes to the development of laminar specificity in ferret visual cortex.

Layer-specific cortical axons are believed to develop precisely from the outset without making exuberant branches in incorrect cortical layers. We tested this hypothesis by following the development of axonal arbors of layer 2/3 pyramidal neurons in ferret visual cortex, which in the adult have dense axonal arbors in layers 2/3 and 5 and not in layer 4. We began our studies at earlier ages and sampled at closer intervals than in previous studies. Our studies reveal that between postnatal day (P) 14 and P18 the initial growth of axonal arbors lacks specificity for layers 2/3 and 5 and involves the formation of a small number of branches incorrectly in layer 4. After P18 the number of incorrect axonal branches in layer 4 decreased, whereas there was extensive axonal growth specifically in layers 2/3 and 5. To further study the mechanisms involved in directing the layer-specific growth and elimination of axonal branches, we studied the development of layer 2/3 neurons in slices kept in culture for 5 d. In these studies the initial imprecise growth at P14-18 followed by precise growth after P18 was mimicked in vitro, but the elimination of incorrect axons from layer 4 did not occur. These studies reveal that most axonal arbors grow precisely in the correct layers, but the initial growth involves the formation of a small number of incorrect axonal branches that are later eliminated. Therefore the mechanisms that regulate axonal growth are not as precise initially as later, and mechanisms must exist to allow selective elimination of axon collaterals from incorrect layers.