Whole cell recording

Figure 1 Schematic diagrams illustrating the patch-clamp technique. (A) Overall setup for isolating single ionic channels in an intact patch of cell membrane. P = patch pipet R = reference microelectrode I = intracellular microelectrode Vp = applied patch potential Em = membrane potential Vm = Em — Vp = potential across the patch A = patch-clamp amplifier. (From Ref. 90.) (B) Five different recording configurations, and procedures used to establish them, (i) Cell attached or intact patch (ii) open cell attached patch (iii) whole cell recording (iv) excised outside-out patch (v) excised inside-out patch. Key i = inside of the cell o = outside of the cell. (Adapted from Ref. 283.)...

Hoffman, N. W., Wuarin, J. P. Dudek, F. E. (1994). Whole-cell recordings of spontaneous synaptic currents in medial preoptic neurons from rat hypothalamic shces mediation by amino acid neurotransmitters. Brain Res. 660, 349-52. 

Alreja M, Aghajanian G. Pacemaker activity of locus coeruleus neurons whole-cell recordings in brain slices show dependence on cAMP and protein kinase A. Brain Res 1991 556 339—343. Shiekhattar R, Aston-Jones G. Modulation of opiate responses in brain noradrenergic neurons by cAMP cascade changes with chronic morphine. Neuroscience 1993 57 879-885. 

The electrophysiological technique used to measure changes in membrane capacitance is the patch clamp [5,6] in the whole-cell recording mode, where the plasma membrane patch in the pipet is ruptured. In another configuration of the patch clamp, the plasma membrane patch is maintained intact. In this case, small currents due to the opening of individual channels can be measured in the membrane patch. The whole-cell patch clamp... 

I/R, inward rectification O/R, outward rectification CPA, cyclopiazonic acid TG, thapsigargin CCh, carbamylcholine CIF, Ca2+ influx factor CAM-ant., calmodulin antagonist W/C, whole-cell recording C/A, cell-attached recording P-W/C, perforated whole-cell recording. 

The isolated oocytes are still surrounded by the vitelline membrane, an inner glycoprotein matrix layer (Yao et al., 2000). This layer does not interfere with whole-cell recording experiments. It can be removed manually however, for patch-clamp experiments requiring the formation of high-resistance seals. 

Selected entries from Methods in Enzymology [vol, page(s)] Analysis, software for, 207, 717 barrier models, 207, 818 closed and open time estimation, 207, 755 data acquisition, 207, 747 modal behavior analysis, 207, 757 multiple channel problem, 207, 756 single-channel [extraction of kinetic information, 207, 765 measurement in tissue slices, 207, 220] synaptic, resolution improvement in patch clamp recording, 207, 216 whole-cell recording in calcium channel, 207, 181 fluctuation analysis, 207, 192. 

The second advantage of this method is its rapidity electrical signals can be detected reliably with a millisecond time resolution. Furthermore, these whole cell recordings can be performed on any cell type or neuronal process in culture, in brain slices as well as in vivo (Margrie et al., 2002). 

As indicated in the previous sections, whole-cell recordings are widely used to detect neurotransmission in multiple settings however, they possess some shortcomings, which hinder their applicability to all forms of neurotransmission. Whole-cell recordings can only be effective to detect fast neurotransmitters that are released... 

Penington NJ, Kelly JS, Fox AP. Whole-cell recordings of inwardly rectifying K+ currents activated by 5-HT1A receptors on dorsal raphe neurones of the adult rat. J Physiol 1993 469 387-405. 

Finally, whole-cell recordings of membrane potential from medium spiny neurons in nucleus accumbens of wild-type and 5-HT1B receptor knockout mice show that 5-HT1B receptor agonists inhibited glutamate-induced EPSC in wild-type, but not knockout, mice (165). 

Jauch, P., Peterson, O.H., Lauger, P. (1986). Electrogenic properties of the sodium-alanine cotransporter in pancreatic acinar cells. I. Tight-seal whole-cell recordings. J. Membr. Biol. 94,99-115. 

An interesting study was performed on cortical slices (from young adult ferrets) with paired whole-cell recording to study the influence of DA on local inhibitory circuits (Gao et al., 2003). In this investigation, DA was found to depress inhibitory transmission between fast-spiking interneurons and pyramidal neurons, but enhanced inhibition between nonfast-spiking interneurons and pyramidal cells. 

The immediate and reversible actions of dopamine are a combination of modulations of individual ion channels. The major ion channels expressed in spiny projection neurons are summarized in Table 2. Current understanding of the properties of these channels is mostly based on whole cell recordings from isolated cells, which have been recently reviewed by Nichola et al. (2000). The role of these channels in whole cell behavior has been studied using intracellular recordings in brain slices or anaesthetized animals. Many of the important cellular properties of spiny projection neurons can be accounted for in terms of ion channel activations occurring at different membrane potentials. 

B.aman In whole cell recordings from Purkinje cells over time, the inactivation curve tends to shift to more negative voltages. 

Horn, R. and Marty, A. (1988) Muscarinic activation of ionic currents measured by a new whole-cell recording method. Journal of General Physiology, 92 145-159. 

Yazulla, S., Studholme, McIntosh, H.H., Ean, S.-H (2000) Cannabinoid receptors on goldfish retinal bipolar cells Electron-microscope immunocytochemistry and whole-cell recordings. Vis. Neurosci. 17, 391 01. 

MiCTOSCOpes A low-power stereomicroscope is sufficient for visualizing tissue and electrode placement for extracellular recordings. A Zeiss Axioscope or other similar microscope equipped with differential interference contrasl/inliared (DIC-IR) optics is necessary for visualized whole-cell recordings. 

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