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In vivo patch clamp recording

Soumya Chatterjee, Heather A Sullivan, Bryan J MacLennan, Ran Xu, YuanYuan Hou, Thomas K Lavin, Nicholas E Lea, Jacob E Michalski, Kelsey R Babcock, Stephan Dietrich, Gillian A Matthews, Anna Beyeler, Gwendolyn G Calhoon, Gordon Glober, Jennifer D Whitesell, Shenqin Yao, Ali Cetin, Julie A Harris, Hongkui Zeng, Kay M Tye, R Clay Reid, Ian R Wickersham
Recombinant rabies viral vectors have proven useful for applications including retrograde targeting of projection neurons and monosynaptic tracing, but their cytotoxicity has limited their use to short-term experiments. Here we introduce a new class of double-deletion-mutant rabies viral vectors that left transduced cells alive and healthy indefinitely. Deletion of the viral polymerase gene abolished cytotoxicity and reduced transgene expression to trace levels but left vectors still able to retrogradely infect projection neurons and express recombinases, allowing downstream expression of other transgene products such as fluorophores and calcium indicators...
March 5, 2018: Nature Neuroscience
Tânia C Gonçalves, Rachid Boukaiba, Jordi Molgó, Muriel Amar, Michel Partiseti, Denis Servent, Evelyne Benoit
The Chinese bird spider huwentoxin-IV (HwTx-IV) is well-known to be a highly potent blocker of NaV 1.7 subtype of voltage-gated sodium (NaV ) channels, a genetically validated analgesic target, and thus promising as a potential lead molecule for the development of novel pain therapeutics. In the present study, the interaction between HwTx-IV and NaV 1.6 channel subtype was investigated using multiscale (from in vivo to individual cell) functional approaches. HwTx-IV was approximatively 2 times more efficient than tetrodotoxin (TTX) to inhibit the compound muscle action potential recorded from the mouse skeletal neuromuscular system in vivo, and 30 times more effective to inhibit nerve-evoked than directly-elicited muscle contractile force of isolated mouse hemidiaphragms...
February 20, 2018: Neuropharmacology
Yuanlong Song, Linlin Gao
The acutely dissociated neurons from the dorsal root ganglia (DRGs) are extensively used. The effects of acute dissociation on the properties of these neurons are, however, not clear. In this study, the action potentials (APs) were recorded from both acutely dissociated and in vivo identified DRG neurons with patch clamp and sharp electrode recording techniques, respectively. We found that acute dissociation slowed both the depolarizing and repolarizing rate of APs, and elongated the AP duration (APD). The lower recording temperature presented in the acutely dissociated neurons contributed to about 10% of these differences...
February 23, 2018: Somatosensory & Motor Research
Jim Sellmeijer, Victor Mathis, Sylvain Hugel, Xu-Hui Li, Qian Song, Qi-Yu Chen, Florent Barthas, Pierre-Eric Lutz, Meltem Karatas, Andreas Luthi, Pierre Veinante, Michel Barrot, Min Zhuo, Ipek Yalcin
Pain associates both sensory and emotional aversive components, and often leads to anxiety and depression when it becomes chronic. Here, we characterized, in a mouse model, the long-term development of these sensory and aversive components as well as anxiodepressive-like consequences of neuropathic pain and determined their electrophysiological impact on the anterior cingulate cortex (ACC, cortical areas 24a/24b). We show that these symptoms of neuropathic pain evolve and recover in different time courses following nerve injury in male mice...
February 20, 2018: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
Jessica Tooley, Lauren Marconi, Jason Bondoc Alipio, Bridget Matikainen-Ankney, Polymnia Georgiou, Alexxai V Kravitz, Meaghan C Creed
BACKGROUND: The ability to appropriately integrate and respond to rewarding and aversive stimuli is essential for survival. The ventral pallidum (VP) plays a critical role in processing both rewarding and aversive stimuli. However, the VP is a heterogeneous structure, and how VP subpopulations integrate into larger reward networks to ultimately modulate these behaviors is not known. We identify a noncanonical population of glutamatergic VP neurons that play a unique role in responding to aversive stimuli and constraining inappropriate reward seeking...
January 12, 2018: Biological Psychiatry
Maria Giese, Julia Gestrich, Azar Massah, Jonas Peterle, HongYing Wei, Monika Stengl
In the Madeira cockroach pigment-dispersing factor-immunoreactive (PDF-ir) neurons innervating the circadian clock, the accessory medulla (AME) in the brain's optic lobes, control circadian behavior. Circadian activity rhythms are entrained to daily light-dark cycles only by compound eye photoreceptors terminating in the lamina and medulla. Still, it is unknown which neurons connect the photoreceptors to the clock to allow for light-entrainment. Here, we characterized by multiple-label immunocytochemistry the serotonin (5-HT)-ir anterior fiber fan and GABA-ir pathways connecting the AME- and optic lobe neuropils...
February 11, 2018: European Journal of Neuroscience
Teresa Guillamon-Vivancos, William A Tyler, Maria Medalla, Wayne Wei-En Chang, Mayumi Okamoto, Tarik F Haydar, Jennifer I Luebke
How the variety of neurons that organize into neocortical layers and functional areas arises is a central question in the study of cortical development. While both intrinsic and extrinsic cues are known to influence this process, whether distinct neuronal progenitor groups contribute to neuron diversity and allocation is poorly understood. Using in vivo genetic fate-mapping combined with whole-cell patch clamp recording, we show that the firing pattern and apical dendritic morphology of excitatory neurons in layer 4 of the barrel cortex are specified in part by their neural precursor lineage...
February 3, 2018: Cerebral Cortex
Mark A Skarsfeldt, Sofia H Bomholtz, Pia R Lundegaard, Angelica Lopez-Izquierdo, Martin Tristani-Firouzi, Bo Hjorth Bentzen
AIM: The zebrafish has emerged as a novel model for investigating cardiac physiology and pathology. The aim of the current study is to investigate the atrium specific ion channels responsible for shaping the atrial cardiac action potential in zebrafish. METHODS: Using quantitative polymerase chain reaction we assessed the expression level of atrium specific potassium channels. The functional role of these channels was studied by patch clamp experiments on isolated atrial and ventricular cardiomyocytes, and by optical mapping of explanted adult zebrafish hearts...
February 7, 2018: Acta Physiologica
Feng Gu, Isabel Parada, Tao Yang, Frank M Longo, David A Prince
Post-traumatic epilepsy is one of the most common and difficult to treat forms of acquired epilepsy worldwide. Currently, there is no effective way to prevent post-traumatic epileptogenesis. It is known that abnormalities of interneurons, particularly parvalbumin-containing interneurons, play a critical role in epileptogenesis following traumatic brain injury. Thus, enhancing the function of existing parvalbumin interneurons might provide a logical therapeutic approach to prevention of post-traumatic epilepsy...
February 2, 2018: Neurobiology of Disease
Shin Hamamoto, Yasuo Mori, Isamu Yabe, Nobuyuki Uozumi
Saccharomyces cerevisiae possesses a transient receptor potential (TRP) channel homolog TRPY1 in its vacuolar membrane, considered to be an ancestral TRP channel. So far, studies have focused on the channel properties of TRPY1 but its regulation and physiological role remained to be elucidated. Here we investigated TRPY1 channel function in vitro and in vivo. Patch-clamp recording of TRPY1 in yeast vacuolar membranes showed that Ca2+ on the lumen side inhibited TRPY1-mediated channel activity, whereas luminal Zn2+ increased the currents...
February 6, 2018: FEBS Journal
Dongze Zhang, Huiyin Tu, Liang Cao, Hong Zheng, Robert L Muelleman, Michael C Wadman, Yu-Long Li
BACKGROUND: Attenuated cardiac vagal activity is associated with ventricular arrhythmogenesis and related mortality in patients with chronic heart failure. Our recent study has shown that expression of N-type Ca2+ channel α-subunits (Cav2.2-α) and N-type Ca2+ currents are reduced in intracardiac ganglion neurons from rats with chronic heart failure. Rat intracardiac ganglia are divided into the atrioventricular ganglion (AVG) and sinoatrial ganglion. Ventricular myocardium receives projection of neuronal terminals only from the AVG...
January 15, 2018: Journal of the American Heart Association
Gong Zhang, Xiao-Hua Wu, Guo-Zhong Xu, Shi-Jun Weng, Xiong-Li Yang, Yong-Mei Zhong
Orexin-A, -B play a crucial role in arousal and feeding by activating two G-protein-coupled receptors: orexin receptor 1 (OX1R) and orexin receptor 2 (OX2R). Orexins, along with orexin receptors, are expressed in retinal neurons, and they have been shown to differentially modulate excitatory AMPA receptors of amacrine and ganglion cells in the inner retina. In this work we report that orexin-B modulates the activity of rod bipolar cells (RBCs) located in the outer retina of rat. Intravitreal injection of orexin-B increased the amplitude of the scotopic electroretinographic b-wave, a reflection of RBC activity, recorded in vivo...
January 8, 2018: Neuropharmacology
Xingxing Liu, Haibo Song, Jun Yang, Cheng Zhou, Yi Kang, Linghui Yang, Jin Liu, Wensheng Zhang
OBJECTIVE: (R)-2-methoxyethyl1-(1-phenylethyl)-1H-imidazole-5-carboxylate hydrochloride (ET-26 HCl) is a novel etomidate analogue. The purpose of this study was to characterize whether ET-26 HCl could retain the superior myocardial performance of etomidate in vivo and in vitro. METHODS: In vivo, the influence of ET-26 HCl and etomidate on the cardiac function of dogs was confirmed using echocardiography and electrocardiogram. In vitro, a Langendorff preparation was used to examine direct myocardial performance in isolated rat hearts, and a whole-cell patch-clamp technique was used to study effects on the human ether-a-go-go-related gene (hERG) channel...
2018: PloS One
Suhasa B Kodandaramaiah, Francisco J Flores, Gregory L Holst, Annabelle C Singer, Xue Han, Emery N Brown, Edward S Boyden, Craig R Forest
The activities of groups of neurons in a circuit or brain region are important for neuronal computations that contribute to behaviors and disease states. Traditional extracellular recordings have been powerful and scalable, but much less is known about the intracellular processes that lead to spiking activity. We present a robotic system, the multipatcher, capable of automatically obtaining blind whole-cell patch clamp recordings from multiple neurons simultaneously. The multipatcher significantly extends automated patch clamping, or 'autopatching', to guide four interacting electrodes in a coordinated fashion, avoiding mechanical coupling in the brain...
January 3, 2018: ELife
Jessica N Peoples, David G Taylor, Alexander N Katchman, Steven N Ebert
Mouse embryos that lack the ability to produce the adrenergic hormones, norepinephrine (NE) and epinephrine (EPI), due to disruption of the dopamine beta-hydroxylase (Dbh-/-) gene inevitably perish from heart failure during mid-gestation. Since adrenergic stimulation is well-known to enhance calcium signaling in developing as well as adult myocardium, and impairments in calcium signaling are typically associated with heart failure, we hypothesized that adrenergic-deficient embryonic hearts would display deficiencies in cardiac calcium signaling relative to adrenergic-competent controls at a developmental stage immediately preceding the onset of heart failure, which first appears beginning or shortly after mouse embryonic day 10...
December 27, 2017: Biochemical and Biophysical Research Communications
Luca Sala, Berend J van Meer, Leon T Tertoolen, Jeroen Bakkers, Milena Bellin, Richard P Davis, Chris N Denning, Michel A Dieben, Thomas Eschenhagen, Elisa Giacomelli, Catarina Grandela, Arne Hansen, Eduard Holman, Monique R Jongbloed, Sarah M Kamel, Charlotte D Koopman, Quentin Lachaud, Ingra Mannhardt, Mervyn P Mol, Diogo Mosqueira, Valeria V Orlova, Robert Passier, Marcelo C Ribeiro, Umber Saleem, Godfrey Smith, Francis L L Burton, Christine L Mummery
Rationale: There are several methods to measure cardiomyocyte (CM) and muscle contraction but these require customized hardware, expensive apparatus and advanced informatics or can only be used in single experimental models. Consequently, data and techniques have been difficult to reproduce across models and laboratories, analysis is time consuming and only specialist researchers can quantify data. Objective: Here we describe and validate an automated, open source software tool (MUSCLEMOTION) adaptable for use with standard laboratory- and clinical imaging equipment that enables quantitative analysis of normal cardiac contraction, disease phenotypes and pharmacological responses...
December 27, 2017: Circulation Research
D Falcón, R González-Montelongo, E Sánchez de Rojas-de Pedro, A Ordóñez, J Ureña, A Castellano
Glucocorticoids are widely used to treat acute and chronic diseases. Unfortunately, their therapeutic use is associated with severe side effects. Glucocorticoids are known to regulate several ion channels in cardiac myocytes, including voltage-dependent Ca2+ channels. Low-voltage-activated T-type Ca2+ channels are expressed in ventricular myocytes during the fetal and perinatal period, but are practically absent in the adult. However, these channels can be re-expressed in adult cardiomyocytes under some pathological conditions...
December 17, 2017: Journal of Steroid Biochemistry and Molecular Biology
Darin Edwards, Frank Sommerhage, Bonnie Berry, Hanna Nummer, Martina Raquet, Brad Clymer, Maria Stancescu, James J Hickman
Microelectrode arrays (MEAs) are innovative tools used to perform electrophysiological experiments for the study of electrical activity and connectivity in populations of neurons from dissociated cultures. Reliance upon neurons derived from embryonic tissue is a common limitation of neuronal/MEA hybrid systems and perhaps of neuroscience research in general, and the use of adult neurons could model fully functional in vivo parameters more closely. Spontaneous network activity was concurrently recorded from both embryonic and adult rat neurons cultured on MEAs for up to 10 weeks in vitro to characterize the synaptic connections between cell types...
December 11, 2017: ACS Biomaterials Science & Engineering
Shogo Tsujikawa, Tadashi Matsuura, Kotaro Hori, Takashi Mori, Miyuki Kuno, Kiyonobu Nishikawa
BACKGROUND: Tricyclic antidepressants (TCAs) are a major cause of fatal drug poisoning due to their cardiotoxicity. Alkalinization by sodium bicarbonate (NaHCO3) administration, the first-line therapy for TCA-induced cardiotoxicity, can occasionally yield insufficient efficacy in severe cases. Because most TCAs are highly lipophilic, lipid emulsion may be more effective than alkalinization. However, it remains to be determined whether lipid emulsion is more beneficial than alkalinization in reversing amitriptyline-induced cardiotoxicity...
December 11, 2017: Anesthesia and Analgesia
Anna R Chambers, Nadia Pilati, Pooja Balaram, Charles H Large, Leonard K Kaczmarek, Daniel B Polley
Higher stages of central auditory processing compensate for a loss of cochlear nerve synapses by increasing the gain on remaining afferent inputs, thereby restoring firing rate codes for rudimentary sound features. The benefits of this compensatory plasticity are limited, as the recovery of precise temporal coding is comparatively modest. We reasoned that persistent temporal coding deficits could be ameliorated through modulation of voltage-gated potassium (Kv) channels that regulate temporal firing patterns...
December 13, 2017: Scientific Reports
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