The cerebral cortex is subdivided into six layers based on morphological features. The supragranular layers 2/3 (L2/3) contain morphologically and genetically diverse populations of neurons, suggesting the existence of discrete classes of cells. In primates and carnivores L2/3 can be subdivided morphologically, but cytoarchitectonic divisions are less clear in rodents. Nevertheless, discrete classes of cells could exist based on their computational requirement, which might be linked to their associated functional microcircuits. Through slice recordings coupled with laser-scanning photostimulation we investigated whether L2/3 of male mouse auditory cortex contains discrete subpopulations of cells with specific functional microcircuits. We use hierarchical clustering on the laminar connection patterns to reveal the existence of multiple distinct classes of L2/3 neurons. The classes of L2/3 neurons are distinguished by the pattern of their laminar and columnar inputs from within A1 and their location within L2/3. Cells in superficial L2 show more extensive columnar integration than deeper L3 cells. Moreover, L3 cells receive more translaminar input from L4. imaging in awake mice revealed that L2 cells had higher bandwidth than L3 cells, consistent with the laminar differences in columnar integration. These results suggest that similar to higher mammals, rodent L2/3 is not a homogenous layer but contains several parallel microcircuits. Layer 2/3 of auditory cortex is functionally diverse. We investigated whether L2/3 cells form classes based on their functional connectivity. We used whole-cell patch-clamp recordings with laser-scanning photostimulation and performed unsupervised clustering on the resulting excitatory and inhibitory connection patterns. Cells within each class were located in different sublaminae. Superficial cells showed wider integration along the tonotopic axis and the amount of L4 input varied with sublaminar location. To identify whether sensory responses varied with sublaminar location, we performed Ca imaging and found that L2 cells were less frequency-selective than L3 cells. Our results show that the diversity of receptive fields in L2/3 is likely due to diversity in the underlying functional circuits.

During the critical period, neuronal connections are shaped by sensory experience. While the basis for this temporarily heightened plasticity remains unclear, shared connections introducing activity correlations likely play a key role. Thus, we investigated the changing intracortical connectivity in primary auditory cortex (A1) over development. In adult, layer 2/3 (L2/3) neurons receive ascending inputs from layer 4 (L4) and also receive few inputs from subgranular layer 5/6 (L5/6). We measured the spatial pattern of intracortical excitatory and inhibitory connections to L2/3 neurons in slices of mouse A1 across development using laser-scanning photostimulation. Before P11, L2/3 cells receive most excitatory input from within L2/3. Excitatory inputs from L2/3 and L4 increase after P5 and peak during P9–16. L5/6 inputs increase after P5 and provide most input during P12–16, the peak of the critical period. Inhibitory inputs followed a similar pattern. Functional circuit diversity in L2/3 emerges after P16. In vivo two-photon imaging shows low pairwise signal correlations in neighboring neurons before P11, which peak at P15–16 and decline after. Our results suggest that the critical period is characterized by high pairwise activity correlations and that transient hyperconnectivity of specific circuits, in particular those originating in L5/6, might play a key role.

SUMMARYLoss of a sensory modality can lead to functional enhancement of the remaining senses. For example, short-term visual deprivations, or dark exposure (DE), can enhance neuronal responses in the auditory cortex to sounds. These enhancements encompass increased spiking rates and frequency selectivity as well as increased spiking reliability. Although we previously demonstrated enhanced thalamocortical transmission after DE, increased synaptic strength cannot account for increased frequency selectivity or reliability. We thus investigated whether other changes in the underlying circuitry contributed to improved neuronal responses. We show that DE can lead to refinement of intra- and inter-laminar connections in the mouse auditory cortex. Moreover, we use a computational model to show that the combination of increased transmission and circuit refinement can lead to increased firing reliability. Thus cross-modal influences can alter the spectral and temporal processing of sensory stimuli by refinement of thalamocortical and intracortical circuits.

Some neurons in the nervous system do not show repetitive firing for steady currents. For time-varying inputs, they fire once if the input rise is fast enough. This property of phasic firing is known as Type III excitability. Type III excitability has been observed in neurons in the auditory brainstem (MSO), which show strong phase-locking and accurate coincidence detection. In this paper, we consider a Hodgkin-Huxley type model (RM03) that is widely-used for phasic MSO neurons and we compare it with a modification of it, showing tonic behavior. We provide insight into the temporal processing of these neuron models by means of developing and analyzing two reduced models that reproduce qualitatively the properties of the exemplar ones. The geometric and mathematical analysis of the reduced models allows us to detect and quantify relevant features for the temporal computation such as nearness to threshold and a temporal integration window. Our results underscore the importance of Type III excitability for precise coincidence detection.

Optical mapping is an emerging single molecule approach for the rapid generation of ordered restriction maps, using fluorescence microscopy. We have improved the size resolution of optical mapping by imaging individual DNA molecules elongated and fixed onto derivatized glass surfaces. Averaged fluorescence intensity and apparent length measurements accurately determined the mass of restriction fragments 800 basepairs long. We have used optical mapping to create ordered restriction maps for lambda clones derived from the mouse pygmy locus.