Alan Woodruff

Chandelier (axoaxonic) cells (ChCs) are a distinct group of GABAergic interneurons that innervate the axon initial segments of pyramidal cells. However, their circuit role and the function of their clearly defined anatomical specificity remain unclear. Recent work has demonstrated that chandelier cells can produce depolarizing GABAergic PSPs, occasionally driving postsynaptic targets to spike. On the other hand, other work suggests that ChCs are hyperpolarizing and may have an inhibitory role. These disparate functional effects may reflect heterogeneity among ChCs. Here, using brain slices from transgenic mouse strains, we first demonstrate that, across different neocortical areas and genetic backgrounds, upper Layer 2/3 ChCs belong to a single electrophysiologically and morphologically defined population, extensively sampling Layer 1 inputs with asymmetric dendrites. Consistent with being a single cell type, we find electrical coupling between ChCs. We then investigate the effect of chandelier cell activation on pyramidal neuron spiking in several conditions, ranging from the resting membrane potential to stimuli designed to approximatein vivomembrane potential dynamics. We find that under quiescent conditions, chandelier cells are capable of both promoting and inhibiting spike generation, depending on the postsynaptic membrane potential. However, duringin vivo-like membrane potential fluctuations, the dominant postsynaptic effect was a strong inhibition. Thus, neocortical chandelier cells, even from within a homogeneous population, appear to play a dual role in the circuit, helping to activate quiescent pyramidal neurons, while at the same time inhibiting active ones.

Chandelier (or axo-axonic) cells are one of the most distinctive types of GABAergic interneurons in the cortex. Although they have traditionally been considered inhibitory neurons, data from rat and human neocortical preparations suggest that chandelier cells have a depolarizing effect on pyramidal neurons at resting membrane potential, and could even activate synaptic chains of neurons. At the same time, recent results from rat hippocampal chandeliers indicate a predominantly inhibitory effect on their postsynaptic targets. To better understand the function of chandelier neurons, we generated Nkx2.1Cre MADM mice, a strain of genetically engineered animals that, by expressing GFP in a subset of neocortical interneurons, enable the identification and targeting of chandelier cells in living brain slices. Using these mice, we characterized the basic electrophysiological properties of a homogeneous population of chandelier neurons from upper layers of somatosensory cortical slices. These chandelier cells have characteristic axon cartridges and stereotypical electrophysiological features, distinguishable from basket cells. To investigate the effect of chandelier cells on target neurons, we performed paired recordings from chandeliers and postsynaptic pyramidal cells. In both perforated patch and cell-attached configurations, chandelier PSPs have in every case a reversal potential that is depolarized from rest. Our results support the idea that chandelier cells depolarize pyramidal neurons and could potentially have an excitatory effect on the network at rest.

Laser microscopy has generally poor temporal resolution, caused by the serial scanning of each pixel. This is a significant problem for imaging or optically manipulating neural circuits, since neuronal activity is fast. To help surmount this limitation, we have developed a “scanless” microscope that does not contain mechanically moving parts. This microscope uses a diffractive spatial light modulator (SLM) to shape an incoming two-photon laser beam into any arbitrary light pattern. This allows the simultaneous imaging or photostimulation of different regions of a sample with three-dimensional precision. To demonstrate the usefulness of this microscope, we perform two-photon uncaging of glutamate to activate dendritic spines and cortical neurons in brain slices. We also use it to carry out fast (60 Hz) two-photon calcium imaging of action potentials in neuronal populations. Thus, SLM microscopy appears to be a powerful tool for imaging and optically manipulating neurons and neuronal circuits. Moreover, the use of SLMs expands the flexibility of laser microscopy, as it can substitute traditional simple fixed lenses with any calculated lens function.