Journal of neuroscience methods
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J. Neurosci. Methods · Jun 2007
The CatWalk method: a detailed analysis of behavioral changes after acute inflammatory pain in the rat.
Experimental pain research is often complicated by the absence of an objective and detailed method to analyze behavioral changes. In the present study, acute pain was induced into the right knee of the rat (n=15) through the injection of 2mg carrageenan (CAR) in saline. A control group received vehicle injection into the knee (n=15). ⋯ These CatWalk parameters were highly correlated with von Frey data and thus representative for the development of mechanical allodynia. Furthermore, detailed CatWalk analysis of the gait (i.e. coordinated interaction between left and right hindlimb) showed very fine, accurate and significant coordination changes in the experimental rats from 4h post-injection. In conclusion, the CatWalk method allows an objective and detailed detection of both pain-induced gait adaptations as well as the development of mechanical allodynia in an acute inflammatory pain model.
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J. Neurosci. Methods · May 2007
Automated optimal detection and classification of neural action potentials in extra-cellular recordings.
Determination of single unit spikes from multiunit spike trains plays a critical role in neurophysiological coding studies which require information about the precise timing of events underlying the neural codes that are the basis of behavior. Searching for optimal spike detection strategies has therefore been the focus of many studies over the past two decades. In this study we describe and implement an algorithm for the optimal real time detection and classification of neural spikes. ⋯ The third step uses these templates for the real time spike detection and classification. In this step the incoming data are projected into a lower dimensional space that is designed to maximally separate the signal from the noise energy. This algorithm provides an accurate estimate of the signal to noise ratio and provides an accurate estimate of spike times and spike shapes.
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J. Neurosci. Methods · Mar 2007
Monitoring synaptic transmission in primary neuronal cultures using local extracellular stimulation.
Various techniques have been applied for the functional analysis of synaptic transmission in cultured neurons. Here, we describe a method of studying synaptic transmission in neurons cultured at high-density from different brain regions such as the cortex, striatum and spinal cord. We use postsynaptic whole-cell recordings to monitor synaptic currents triggered by presynaptic action potentials that are induced by brief stimulations with a nearby extracellular bipolar electrode. ⋯ We show that the size and kinetics of pharmacologically isolated inhibitory postsynaptic currents triggered by single action potentials or stimulus trains depend on the Ca2+ concentration, temperature and stimulation frequency. This method can be applied to study synaptic transmission in wildtype neurons infected with lentiviruses encoding various components of presynaptic release machinery, or in neurons from genetically modified mice, for example neurons carrying floxed genes in which gene expression can be acutely ablated by expression of Cre recombinase. The preparation described in this paper should be useful for analysis of synaptic transmission in inter-neuronal synapses formed by different types of neurons.
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J. Neurosci. Methods · Mar 2007
High-field diffusion tensor imaging of mouse brain in vivo using single-shot STEAM MRI.
Information about the microstructural organization of cerebral white matter that is accessible by magnetic resonance diffusion tensor imaging (DTI) gains increasing importance for studies of animal brain. Particular challenges occur for in vivo conditions as well as at high magnetic fields. ⋯ When compared to studies at 2.35 T, half Fourier DW STEAM MRI at 7 T yielded a substantial gain in signal-to-noise ratio (SNR) that could be invested either in a reduction of the measurement time or an increase of the spatial resolution. Thus, for a measurement time of 3h, DTI with a voxel size of 117 microm x 117 microm x 720 microm not only resulted in high-quality maps of the fractional anisotropy and main diffusion direction (MDD), but also allowed for fiber tracking of major mouse brain structures in vivo.
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J. Neurosci. Methods · Mar 2007
Low-density neuronal networks cultured using patterned poly-l-lysine on microelectrode arrays.
Synaptic activity recorded from low-density networks of cultured rat hippocampal neurons was monitored using microelectrode arrays (MEAs). Neuronal networks were patterned with poly-l-lysine (PLL) using microcontact printing (microCP). Polydimethysiloxane (PDMS) stamps were fabricated with relief structures resulting in patterns of 2 microm-wide lines for directing process growth and 20 microm-diameter circles for cell soma attachment. ⋯ Evoked responses of selected portions of the networks were produced by stimulation of specific electrode sites. In addition, the neuronal excitability of the network was increased by the bath application of high K(+) (10-12 mM). Application of DNQX, an AMPA antagonist, blocked all spontaneous activity, suggesting that the activity is excitatory and mediated through glutamate receptors.