Whole-cell patch clamp recordings

Figure 13.3 Whole-cell patch-clamp recordings of excitatory postsynaptic currents (EPSCs) from dorsal horn neurons of rat (prenatal P2-13) spinal cord slices. The normal evoked EPSC of about 160pA obtained by focal stimulation of nearby tissue was dramatically reduced by addition of a cocktail (CABS) of CNQX 10 pM, D-APV 50 pM, bicuculline 10 pM and strychnine 5 pM to block glutamate, GABAa and glycine receptors. The small residual EPSC shown was blocked by the ATP P2 receptor antagonist suramin and is therefore probably mediated by released ATP. (Prom Bardoni et al. 1997 and reproduced by permission of the Journal of Neuroscience)...

Cui, C., Smith, D.O. and Adler, J. (1995) Characterization of mechanosensitive channels in Eschericia cofi cytoplasmic cell membrane by whole-cell patch clamp recording. Journal of Membrane Biology 144 31-42... 

Cepeda C, Chandler SH, Shumate LW, Levine MS (1995) Persistent Na+ conductance in medium-sized neostriatal neurons characterization using infrared videomicroscopy and whole cell patch-clamp recordings. J Neurophysiol 74 1343—1348. 

Fertig N, Blick RH, Behrends JC. Whole cell patch clamp recording performed on a planar glass chip. Biophys. J. 2002 82 3056-3062. 

Figure 2. A. Recording of calcium variations in a photocyte cluster of O. aranea. (i) 200mM KCl stimulation, ( (>) rinsing with ASW. Values are expressed as a ratio between Ca bound fura-2 and free fura-2. B. Whole cell patch-clamp recording on O. californica photocyte. Stimulation protocol holding potential of-lOOmV, steps of 25mV from -lOOmV to -i-lOOmV (a) outward current, (b) inward current...

A recent whole-cell patch-clamp recording smdy (Gu et al. 2008 Fig. 4) has further demonsttated that a brief application of acetylcholine (3, 10, 30, and 100pM,... 

Whole-cell patch-clamp recordings from cultured hippocampal neurones have been used to further characterize the action of 4-PIOL [31] (Fig. 10). The action of 4-PIOL was compared with that of the GABAa agonist isoguvacine. 

Fig. 10. Whole-cell patch-clamp recording from a hippocampal neurone. Holding potential was -60 mV with -10 mV command potentials superimposed to monitor membrane conductance. Drugs were applied in the vicinity of the neurone by a multi-barrel perfusion pipette. The response to isoguvacine was reduced by simultaneous application of 4-PIOL to a value slightly higher than the intrinsic agonist response to 4-PIOL alone. In contrast to the response to 20 pM isoguvacine alone the responses with 4-PIOL present did not show desensitization However, the responses to 10 pM isoguvacine, comparable in strength to the 1 mM 4-PIOL response, did not show desensitization either.

Whole-cell patch clamp recordings are used to investi-... 

Figure 15.1. Drosophila embryonic neuromuscular preparation. Top panel) Dissected Drosophila embryo viewed with a scanning electron microscope (SEM) anterior (A) to the left, posterior (P) to the right. The prominent CNS lies along the ventral midline. Bottom panel) Schematic drawing of the four ventral longitudinal muscles in one segment. The CNS, peripheral nerve, and NMJ on these four muscles are drawn. The typical recording configuration involves whole-cell patch-clamp recording from muscle 6 and suction-electrode stimulation of the peripheral nerve.

The main procedure to relate the electrophysiological properties of cortical neurons to their morphological features is represented by the whole-cell patch clamp recording on slices maintained in vitro. The detailed description of this procedure is beyond the purpose of the present chapter. However, a brief overview of the main steps involved is given below. 

Since the neurons targeted by whole-cell patch clamp recording were also filled with biocytin (0.2 %) we were also able to identify the shape and position of these P -GFP neurons within the cortical network. We used streptavidin conjugated to a red fluorescent probe (Alexa 568). Streptavidin reacts directly with biotin (biocytin) within the filled neuron, labeling only the filled neuron in the red fluorescence channel of the wide-field microscope. Typically we obtained recovery percentages of 80-100 % of neurons using the protocol below. 

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