Efficient Method for Whole-Cell Recording in Freely Moving Rodents Using Ultraviolet-Cured Collar-Based Pipette Stabilization

2017 ◽  
Vol 2017 (4) ◽  
pp. pdb.prot095810 ◽  
Author(s):  
Doyun Lee ◽  
Albert K. Lee
Keyword(s):  
Efficient Method ◽  
Whole Cell ◽  
Whole Cell Recording ◽  
Cell Recording ◽  
Freely Moving

Whole cell recording from neurons in slices of reptilian and mammalian cerebral cortex

1989 ◽  
Vol 30 (3) ◽  
pp. 203-210 ◽  
Author(s):  
Mark G. Blanton ◽  
Joseph J. Lo Turco ◽  
Arnold R. Kriegstein
Keyword(s):  
Cerebral Cortex ◽  
Whole Cell ◽  
Whole Cell Recording ◽  
Cell Recording

scholarly journals Knockouts reveal overlapping functions of M2 and M4 muscarinic receptors and evidence for a local glutamatergic circuit within the laterodorsal tegmental nucleus

2012 ◽  
Vol 108 (10) ◽  
pp. 2751-2766 ◽  
Author(s):  
Kristi A. Kohlmeier ◽  
Masaru Ishibashi ◽  
Jürgen Wess ◽  
Martha E. Bickford ◽  
Christopher S. Leonard
Keyword(s):  
Acetylcholine Receptors ◽  
Brain Slices ◽  
Cholinergic Neurons ◽  
Muscarinic Acetylcholine Receptors ◽  
Feedback Signal ◽  
Laterodorsal Tegmental Nucleus ◽  
Membrane Hyperpolarization ◽  
Whole Cell ◽  
Whole Cell Recording ◽  
Cell Recording

Cholinergic neurons in the laterodorsal tegmental (LDT) and peduncolopontine tegmental (PPT) nuclei regulate reward, arousal, and sensory gating via major projections to midbrain dopamine regions, the thalamus, and pontine targets. Muscarinic acetylcholine receptors (mAChRs) on LDT neurons produce a membrane hyperpolarization and inhibit spike-evoked Ca2+ transients. Pharmacological studies suggest M2 mAChRs are involved, but the role of these and other localized mAChRs (M1--M4) has not been definitively tested. To identify the underlying receptors and to circumvent the limited receptor selectivity of available mAChR ligands, we used light- and electron-immunomicroscopy and whole cell recording with Ca2+ imaging in brain slices from knockout mice constitutively lacking either M2, M4, or both mAChRs. Immunomicroscopy findings support a role for M2 mAChRs, since cholinergic and noncholinergic LDT and pedunculopontine tegmental neurons contain M2-specific immunoreactivity. However, whole cell recording revealed that the presence of either M2 or M4 mAChRs was sufficient, and that the presence of at least one of these receptors was required for these carbachol actions. Moreover, in the absence of M2 and M4 mAChRs, carbachol elicited both direct excitation and barrages of spontaneous excitatory postsynaptic potentials (sEPSPs) in cholinergic LDT neurons mediated by M1 and/or M3 mAChRs. Focal carbachol application to surgically reduced slices suggest that local glutamatergic neurons are a source of these sEPSPs. Finally, neither direct nor indirect excitation were knockout artifacts, since each was detected in wild-type slices, although sEPSP barrages were delayed, suggesting M2 and M4 receptors normally delay excitation of glutamatergic inputs. Collectively, our findings indicate that multiple mAChRs coordinate cholinergic outflow from the LDT in an unexpectedly complex manner. An intriguing possibility is that a local circuit transforms LDT muscarinic inputs from a negative feedback signal for transient inputs into positive feedback for persistent inputs to facilitate different firing patterns across behavioral states.

scholarly journals Compensation of physiological motion enables high-yield whole-cell recording in vivo

2020 ◽  
Author(s):  
William M. Stoy ◽  
Bo Yang ◽  
Ali Kight ◽  
Nathaniel C. Wright ◽  
Peter Y. Borden ◽  
...  
Keyword(s):  
Single Cells ◽  
Strong Dependence ◽  
High Yield ◽  
Patch Clamp Recording ◽  
Gold Standard Method ◽  
Whole Cell ◽  
Whole Cell Recording ◽  
Cell Recording ◽  
Living Mouse

1.1.1AbstractWhole-cell patch-clamp recording in vivo is the gold-standard method for measuring subthreshold electrophysiology from single cells during behavioural tasks, sensory stimulations, and optogenetic manipulation. However, these recordings require a tight, gigaohm resistance, seal between a glass pipette electrode’s aperture and a cell’s membrane. These seals are difficult to form, especially in vivo, in part because of a strong dependence on the distance between the pipette aperture and cell membrane. We elucidate and utilize this dependency to develop an autonomous method for placement and synchronization of pipette’s tip aperture to the membrane of a nearby, moving neuron, which enables high-yield seal formation and subsequent recordings in the deep in the brain of the living mouse, in the thalamus. This synchronization procedure nearly doubles the reported gigaseal yield in the thalamus (>3 mm below the pial surface) from 26% (n=17/64) to 48% (n=32/66). Whole-cell recording yield improved from 10% (n = 9/88) to 24% (n=18/76) when motion compensation was used during the gigaseal formation. As an example of its application, we utilized this system to investigate the role of the sensory environment and ventral posterior medial region (VPM) projection synchrony on intracellular dynamics in the barrel cortex. This method results in substantially greater subcortical whole-cell recording yield than previously reported and thus makes pan-brain whole-cell electrophysiology practical in the living mouse brain.

scholarly journals Progesterone Attenuates Allodynia of Inflamed Temporomandibular Joint through Modulating Voltage-Gated Sodium Channel 1.7 in Trigeminal Ganglion

2020 ◽  
Vol 2020 ◽  
pp. 1-11
Author(s):  
Rui-Yun Bi ◽  
Xiao-Yu Zhang ◽  
Peng Zhang ◽  
Yun Ding ◽  
Ye-Hua Gan
Keyword(s):  
Protein Expression ◽  
Sodium Channel ◽  
Temporomandibular Joint ◽  
Trigeminal Ganglion ◽  
Ovariectomized Rats ◽  
Sodium Currents ◽  
Whole Cell ◽  
Whole Cell Recording ◽  
Cell Recording ◽  
Mrna And Protein Expression

Background. Women with temporomandibular disorders (TMDs) experience some amelioration of pain during pregnancy. Progesterone increases dramatically and steadily during pregnancy. Sodium channel 1.7 (Nav1.7) plays a prominent role in pain perceptions, as evidenced by deletion of Nav1.7 alone leading to a complete loss of pain. In a previous study, we showed that Nav1.7 in trigeminal ganglion (TG) is involved in allodynia of inflamed temporomandibular joint (TMJ). Whether progesterone modulates allodynia of inflamed TMJ through Nav1.7 in TG remains to be investigated. Methods. The effects of progesterone on sodium currents of freshly isolated TG neurons were examined using whole-cell recording. Female rats were ovariectomized and treated with increasing doses of progesterone for 10 days. Complete Freund’s adjuvant was administered intra-articularly to induce TMJ inflammation. TMJ nociceptive responses were evaluated by head withdrawal thresholds. Real-time PCR and Western blotting were used to examine Nav1.7 mRNA and protein expression in TG. Immunohistofluorescence was used to examine the colocalization of progesterone receptors (PRα/β) and Nav1.7 in TG. Results. Whole-cell recording showed that progesterone could attenuate sodium currents. Moreover, progesterone dose-dependently downregulated Nav1.7 mRNA expression and reduced the sensitivity of TMJ nociception in ovariectomized rats. Furthermore, treatment with progesterone attenuated allodynia of inflamed TMJ in a dose-dependent manner and repressed inflammation-induced Nav1.7 mRNA and protein expression in ovariectomized rats. The progesterone receptor antagonist, RU-486, partially reversed the effect of progesterone on allodynia of inflamed TMJ and TMJ inflammation-induced Nav1.7 mRNA and protein expression. Conclusion. Progesterone, by modulating trigeminal ganglionic Nav1.7, may represent a promising agent to prevent allodynia of inflamed TMJ.

scholarly journals Chemosensory responses in isolated olfactory receptor neurons from Necturus maculosus.

1992 ◽  
Vol 99 (3) ◽  
pp. 415-433 ◽  
Author(s):  
V E Dionne
Keyword(s):  
Olfactory Receptor ◽  
Membrane Conductance ◽  
Input Resistance ◽  
Primary Response ◽  
Olfactory Receptor Neurons ◽  
Whole Cell ◽  
Whole Cell Recording ◽  
Necturus Maculosus ◽  
Cell Recording ◽  
Receptor Neurons

Olfactory receptor neurons were isolated without enzymes from the mudpuppy, Necturus maculosus, and tested for chemosensitivity. The cells responded to odorants with changes in firing frequency and alterations in excitability that were detected with tight-seal patch electrodes using on-cell and whole-cell recording conditions. Chemosensitive cells exhibited two primary response characteristics: excitation and inhibition. Both types of primary response were observed in different cells stimulated by mixtures of amino acids as well as by the single compound L-alanine, suggesting that there may be more than one transduction pathway for some odorants. Using the normal whole-cell recording method, the chemosensitivity of competent cells washed out rapidly; a resistive whole-cell method was used to record odorant responses under current-clamp conditions. In response to chemical stimulation, excitability appeared to be modulated in several different ways in different cells: odorants induced hyperpolarizing or depolarizing receptor potentials, elicited or inhibited transient, rhythmic generator potentials, and altered excitability without changing the membrane potential or input resistance. These effects suggest that olfactory transduction is mediated through at least three different pathways with effects on four or more components of the membrane conductance. Polychotomous pathways such as these may be important for odor discrimination and for sharpening the "odor image" generated in the olfactory epithelium.

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