The ability of the brain to adapt to environmental demands implies that neurons can change throughout life. The extent to which single neurons actually change remains largely unstudied, however. To evaluate how functional properties of single neurons change over time, we devised a way to perform in vivo time-lapse electrophysiological recordings from the exact same neuron. We monitored the contralateral and ipsilateral sensory-evoked spiking activity of individual L2/3 neurons from the somatosensory cortex of mice. At the end of the first recording session, we electroporated the neuron with a DNA plasmid to drive GFP expression. Then, 2 wk later, we visually guided a recording electrode in vivo to the GFP-expressing neuron for the second time. We found that contralateral and ipsilateral evoked responses (i.e., probability to respond, latency, and preference), and spontaneous activity of individual L2/3 pyramidal neurons are stable under control conditions, but that this stability could be rapidly disrupted. Contralateral whisker deprivation induced robust changes in sensoryevoked response profiles of single neurons. Our experiments provide a framework for studying the stability and plasticity of single neurons over long time scales using electrophysiology.electroporation | two-photon imaging I t is well established that the morphology and response properties of neurons are highly dynamic during development. Immature cortical neurons are particularly amenable to change during specific stages of development, when spontaneous activity and sensory experience shape their morphology and physiology (1-4). Once formed and stabilized, neurons are thought to maintain some aspects of this plasticity during adulthood, albeit at a lower level (5-7); however, the extent to which single-neuron physiology changes or remains stable in the adult mammalian brain is poorly documented.Limitations to the study of single-neuron plasticity stem from technical difficulties as well as from the inherent heterogeneity of single neurons in a circuit. Fine changes, like those that underlie functional plasticity, can go undetected due to data averaging and homeostatic events that may cancel out each other. As a result, plasticity is made experimentally more tractable by studying populations after gross manipulations like sensory deprivation or injury. For example, receptive fields of neurons across adult rodent cortices are known to shift in response to different paradigms of sensory input manipulation (8-14). Whether and how other forms of plasticity, such as learning and memory, mark their signature on single neurons is more difficult to deduce from gross manipulation studies. One way to overcome some of these difficulties is through experiments based on time-lapse imaging and electrophysiology (14-18). However, the ability to measure fine temporal physiological events of identifiable single neurons over long time scales remains limited, especially with electrophysiology.To establish a method to follow the electrophysiology of single neuron...

SUMMARY Learning to predict the component in the sensory information resulting from the organism's own activity enables it to respond appropriately to unexpected stimuli. For example, the elasmobranch dorsal octavolateral nucleus (DON) can apparently extract the unexpected component (i.e. generated by nearby organisms) from the incoming electrosensory signals. Here we introduce a novel and unique experimental approach that combines the advantages of in vitro preparations with the integrity of in vivo conditions. In such an experimental system one can study, under control conditions, the cellular and network mechanisms that underlie cancellation of expected sensory inputs. Using extracellular and intracellular recordings we compared the dynamics and spatiotemporal organization of the electrosensory afferent nerve and parallel fiber inputs to the DON. The afferent nerve has a low threshold and high conduction velocity; a stimulus that recruits a small number of fibers is sufficient to activate the principal neurons. The excitatory postsynaptic potential in the principal cells evoked by afferent nerve fibers has fast kinetics that efficiently reach the threshold for action potential. In contrast, the parallel fibers have low conduction velocity, high threshold and extensive convergence on the principal neurons of the DON. The excitatory postsynaptic response has slow kinetics that provides a wide time window for integration of inputs. The highly efficient connection between the afferent nerve and the principal neurons in the DON indicates that filtration occurring in the DON cannot be mediated simply by summation of the parallel fibers' signals with the afferent sensory signals. Hence we propose that the filtering may be mediated via secondary neurons that adjust the principal neurons'sensitivity to afferent inputs.

Exercise and inherited factors both affect recovery from stroke and head injury, but the underlying mechanisms and interconnections between them are yet unknown. Here, we report that similar cation channels mediate the protective effect of exercise and specific genetic background in a kainate injection model of cerebellar stroke. Microinjection to the cerebellum of the glutamatergic agonist, kainate, creates glutamatergic excito-toxicity characteristic of focal stroke, head injury or alcoholism. Inherited protection and prior exercise were both accompanied by higher cerebellar expression levels of the Kir6.1 ATP-dependent potassium channel in adjacent Bergmann glia, and voltage-gated KVbeta2 and cyclic nucleotide-gated cation HCN1 channels in basket cells. Sedentary FVB/N and exercised C57BL/6 mice both expressed higher levels of these cation channels compared to sedentary C57BL/6 mice, and were both found to be less sensitive to glutamate toxicity. Moreover, blocking ATP-dependent potassium channels with Glibenclamide enhanced kainate-induced cell death in cerebellar slices from the resilient sedentary FVB/N mice. Furthermore, exercise increased the number of acetylcholinesterase-positive fibres in the molecular layer, reduced cerebellar cytokine levels and suppressed serum acetylcholinesterase activity, suggesting anti-inflammatory protection by enhanced cholinergic signalling. Our findings demonstrate for the first time that routine exercise and specific genetic backgrounds confer protection from cerebellar glutamatergic damages by similar molecular mechanisms, including elevated expression of cation channels. In addition, our findings highlight the involvement of the cholinergic anti-inflammatory pathway in insult-inducible cerebellar processes. These mechanisms are likely to play similar roles in other brain regions and injuries as well, opening new venues for targeted research efforts.

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