Genetic manipulation, whole-cell recordings and functional imaging of the sensorimotor cortex of behaving mice

Sensory processing, sensorimotor integration and motor control are amongst the most basic functions of the brain and yet our understanding of how the underlying neuronal networks operate and contribute to behaviour is very limited. The relative simplicity of the mouse whisker sensorimotor system is helpful for detailed quantitative analyses of motor control and perception during active sensory processing. Recent technical advances now allow the measurement of membrane potential in awake-behaving mice, using whole-cell recordings and voltage-sensitive dye imaging. With these recording techniques, it is possible to directly correlate neuronal activity with behaviour. However, in order to obtain causal evidence for the specific contributions of different neuronal networks to behaviour, it is critical to manipulate the system in a highly controlled manner. Advances in molecular neurobiology, gene delivery and mouse genetics provide techniques capable of layer, column and cell-type specific control of gene expression in the mouse neocortex. Over the next years, we anticipate considerable advances in our understanding of brain function through measuring and manipulating neuronal activity with unprecedented precision to probe the molecular and synaptic mechanisms underlying simple forms of active sensory perception and associative learning.

Long-range connectivity of mouse primary somatosensory barrel cortex

Rachel Aronoff, Céline Matéo, Ferenc Mátyás, Carl Petersen, Bernard Schneider

The primary somatosensory barrel cortex processes tactile vibrissae information, allowing rodents to actively perceive spatial and textural features of their immediate surroundings. Each whisker on the snout is individually represented in the neocortex by an anatomically identifiable 'barrel' specified by the segregated termination zones of thalamocortical axons of the ventroposterior medial nucleus, which provide the primary sensory input to the neocortex. The sensory information is subsequently processed within local synaptically connected neocortical microcircuits, which have begun to be investigated in quantitative detail. In addition to these local synaptic microcircuits, the excitatory pyramidal neurons of the barrel cortex send and receive long-range glutamatergic axonal projections to and from a wide variety of specific brain regions. Much less is known about these long-range connections and their contribution to sensory processing. Here, we review current knowledge of the long-range axonal input and output of the mouse primary somatosensory barrel cortex. Prominent reciprocal projections are found between primary somatosensory cortex and secondary somatosensory cortex, motor cortex, perirhinal cortex and thalamus. Primary somatosensory barrel cortex also projects strongly to striatum, thalamic reticular nucleus, zona incerta, anterior pretectal nucleus, superior colliculus, pons, red nucleus and spinal trigeminal brain stem nuclei. These long-range connections of the barrel cortex with other specific cortical and subcortical brain regions are likely to play a crucial role in sensorimotor integration, sensory perception and associative learning.

Wiley-Blackwell2010

Internal brain state regulates membrane potential synchrony in barrel cortex of behaving mice

Carl Petersen, James Poulet

Internal brain states form key determinants for sensory perception, sensorimotor coordination and learning. A prominent reflection of different brain states in the mammalian central nervous system is the presence of distinct patterns of cortical synchrony, as revealed by extracellular recordings of the electroencephalogram, local field potential and action potentials. Such temporal correlations of cortical activity are thought to be fundamental mechanisms of neuronal computation. However, it is unknown how cortical synchrony is reflected in the intracellular membrane potential (V(m)) dynamics of behaving animals. Here we show, using dual whole-cell recordings from layer 2/3 primary somatosensory barrel cortex in behaving mice, that the V(m) of nearby neurons is highly correlated during quiet wakefulness. However, when the mouse is whisking, an internally generated state change reduces the V(m) correlation, resulting in a desynchronized local field potential and electroencephalogram. Action potential activity was sparse during both quiet wakefulness and active whisking. Single action potentials were driven by a large, brief and specific excitatory input that was not present in the V(m) of neighbouring cells. Action potential initiation occurs with a higher signal-to-noise ratio during active whisking than during quiet periods. Therefore, we show that an internal brain state dynamically regulates cortical membrane potential synchrony during behaviour and defines different modes of cortical processing.

Nature Publishing Group2008

Uncovering the neural encoding of behavior in the adult Drosophila motor system

Chin-Lin Chen

One of the most important goals in neuroscience research has always been to understand how animals control their behavior. However, the long focus on the role of brain neurons in behavioral control might be missing the full story. In fact, brain-wide fluctuations in neural activity are highly correlated to the behavioral state, suggesting a closed-loop system where the behavioral command and the body state interact to interpret sensory inputs and dynamically guide action selection. However, to date, little is known about how and where information about the behavioral state is generated. In principle, information about the behavioral state is a relatively high-level encoding of the body state derived from the integration of signals from primary sensors and the local network. Local motor circuits residing in the spinal cord of mammals or the ventral nerve cord (VNC) of insects may give rise to the behavioral state and pass to the brain via brain-projecting interneurons---termed ascending neurons (ANs). However, what information ANs carry and where this information originates is still a mystery due to the technical challenges in measuring these signals in behaving animals. Therefore, it is critical to develop tools to track neuron identity and record neuronal activity during behavior to crack the functional network. In this work, a method is introduced for imaging the motor circuit in the VNC of Drosophila melanogaster. This preparation demonstrates how to retain fly limb function after dissection and allows simultaneous recording of a tethered fly's behavior and its neural activity in the VNC. Incorporating this methodology with other approaches can verify the conclusions from the experiments such as those manipulating neuronal excitability, and motivate further investigation into neuronal function in specific types of behavior. Since it has long been theorized that ANs are the cellular basis of the behavioral state information sent in the brain, the technique introduced in this work was designed to resolve questions relating to what type of information ANs convey. Here, a large-scale functional screen was performed to relate neural activity, behavior, and neuronal morphology for 247 genetically identified ANs in the adult fly. This is the first large-scale screen of neural activity in a behaving animal, and this work uncovered three fundamental features of AN populations. First, rather than low-level limb movements, ANs encode high-level behaviors like walking, grooming, and resting. Second, ANs project nearly exclusively to an integrative sensory center (AVLP) and an action selection hub (GNG). Remarkably, the encodings projected to these distinct areas reflect their potential roles in contextualizing sensory cues (AVLP) and guiding future actions (GNG). Third, the behavioral encodings of ANs are closely linked to their motor system patterning. In conclusion, this work provides an indispensable tool to study the motor system beyond the brain network by overcoming a major technical barrier in in-vivo recording of the VNC of a behaving fly. This tool was used to help resolve what is encoded by ANs as a higher-level behavioral state, suggesting that the primary cellular origin of the behavioral information that is used to modulate brain function comes from the VNC. Overall, the technique and the findings on AN encoding should inspire follow-up studies to uncover additional neural mechanisms used in adaptive behavior generation.

EPFL2022

Uncovering the neural encoding of behavior in the adult Drosophila motor system

Chin-Lin Chen

EPFL2022