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Whole cell patch clamp

Jun-Bin Yin, Ya-Cheng Lu, Ban Feng, Zhen-Yu Wu, Ying-Biao Chen, Ting Zhang, Zhen-Zhen Kou, Ming-Ming Zhang, Han Zhang, Jin-Lian Li, Hui Li, Tao Chen, Yu-Lin Dong, Yun-Qing Li
BACKGROUND/AIMS: Spinal dorsal horn (SDH) is one of the most important regions for analgesia produced by endomorphin-2 (EM2), which has a higher affinity and specificity for the µ-opioid receptor (MOR) than morphine. Many studies have focused on substantia gelatinosa (SG, lamina II) neurons to elucidate the cellular basis for its antinociceptive effects. However, the complicated types and local circuits of interneurons in the SG make it difficult to understand the real effects of EM2...
March 15, 2018: Neuro-Signals
Hironori Sagawa, Shinsuke Hoshino, Kengo Yoshioka, Wei-Guang Ding, Mariko Omatsu-Kanbe, Masao Nakagawa, Yoshihiro Maruo, Hiroshi Matsuura
BACKGROUND: In the clinical setting, verapamil is contraindicated in neonates and infants because of the perceived risk of hypotension or bradyarrhythmia. However, it remains unclear whether there is an age-dependent difference in the sensitivity of cardiac L-type Ca2+ channel current (ICa,L ) to inhibition by verapamil. METHODS: Ventricular myocytes were enzymatically dissociated from the hearts of 6 different age groups (0, 7, 14, 21, 28 days and 10-15 weeks) of mice using a similar Langendorff-perfusion method...
March 19, 2018: Pediatric Research
Guang-Zhe Huang, Mutsuo Taniguchi, Ye-Bo Zhou, Jing-Ji Zhang, Fumino Okutani, Yoshihiro Murata, Masahiro Yamaguchi, Hideto Kaba
The formation of mate recognition memory in mice is associated with neural changes at the reciprocal dendrodendritic synapses between glutamatergic mitral cell (MC) projection neurons and GABAergic granule cell (GC) interneurons in the accessory olfactory bulb (AOB). Although noradrenaline (NA) plays a critical role in the formation of the memory, the mechanism by which it exerts this effect remains unclear. Here we used extracellular field potential and whole-cell patch-clamp recordings to assess the actions of bath-applied NA (10 µM) on the glutamatergic transmission and its plasticity at the MC-to-GC synapse in the AOB...
April 2018: Learning & Memory
Robert Steger, Lauren Blachorsky, Qizong Yang, Joshua C Brumberg
The properties of specific cortical cell types enable greater understanding of how cortical microcircuits process and transmit sensory, motor, and cognitive information. Previous reports have characterized the intrinsic properties of the inverted pyramidal cell (IPC) where the most prominent dendrite is orientated towards the cortical white matter. Using whole cell patch clamp recordings from rat and mouse somatosensory cortex in conjunction with electric microstimulation of the white matter we characterized the synaptic inputs onto IPCs and the more common upright pyramidal cell (UPC) in the infragranular layers...
March 13, 2018: Somatosensory & Motor Research
Hyemi Bae, Jeongyoon Choi, Young-Won Kim, Donghee Lee, Jung-Ha Kim, Jae-Hong Ko, Hyoweon Bang, Taeho Kim, Inja Lim
This study investigated the expression of voltage-gated K⁺ (KV ) channels in human cardiac fibroblasts (HCFs), and the effect of nitric oxide (NO) on the KV currents, and the underlying phosphorylation mechanisms. In reverse transcription polymerase chain reaction, two types of KV channels were detected in HCFs: delayed rectifier K⁺ channel and transient outward K⁺ channel. In whole-cell patch-clamp technique, delayed rectifier K⁺ current (IK ) exhibited fast activation and slow inactivation, while transient outward K⁺ current (Ito ) showed fast activation and inactivation kinetics...
March 12, 2018: International Journal of Molecular Sciences
Sha Yan, Pengfei Huang, Ying Wang, Xiongzhi Zeng, Yiya Zhang
Spider venoms are known to contain various toxins that are used as an effective means to capture their prey or to defend themselves against predators. An investigation of the properties of Ornithoctonus huwena (O.huwena) crude venom found that the venom can block neuromuscular transmission of isolated mouse phrenic nerve-diaphragm and sciatic nerve-sartorius preparations. However, little is known about its electrophysiological effects on cardiac myocytes. In this study, electrophysiological activities of ventricular myocytes were detected by 100 μg/mL venom of O...
March 13, 2018: Channels
Agnese Secondo, Alba Esposito, Tiziana Petrozziello, Francesca Boscia, Pasquale Molinaro, Valentina Tedeschi, Anna Pannaccione, Roselia Ciccone, Natascia Guida, Gianfranco Di Renzo, Lucio Annunziato
Nuclear envelope (NE) is a Ca2+ -storing organelle controlling neuronal differentiation through nuclear Ca2+ concentrations ([Ca2+ ]n ). However, how [Ca2+ ]n regulates this important function remains unknown. Here, we investigated the role of the nuclear form of the Na+ /Ca2+ exchanger 1(nuNCX1) during the different stages of neuronal differentiation and the involvement of PTEN/PI3'K/Akt pathway. In neuronal cells, nuNCX1 was detected on the inner membrane of the NE where protein expression and activity of the exchanger increased during NGF-induced differentiation...
December 2018: Cell Death Discovery
Mickaël F El Hachmane, Charlotta S Olofsson
Ca2+ impacts a large array of cellular processes in every known cell type. In the white adipocyte, Ca2+ is involved in regulation of metabolic processes such as lipolysis, glucose uptake and hormone secretion. Although the importance of Ca2+ in control of white adipocyte function is clear, knowledge is still lacking regarding the control of dynamic Ca2+ alterations within adipocytes and mechanisms inducing intracellular Ca2+ changes remain elusive. Own work has recently demonstrated the existence of store-operated Ca2+ entry (SOCE) in lipid filled adipocytes...
March 7, 2018: Biochemical and Biophysical Research Communications
Paula P Perissinotti, María Celeste Rivero-Echeto, Edgar Garcia-Rill, Verónica Bisagno, Francisco J Urbano
Leptin is an adipose-derived hormone that controls appetite and energy expenditure. Leptin receptors are expressed on extra-hypothalamic ventrobasal (VB) and reticular thalamic (RTN) nuclei from embryonic stages. Here, we studied the effects of pressure-puff, local application of leptin on both synaptic transmission and action potential properties of thalamic neurons in thalamocortical slices. We used whole-cell patch-clamp recordings of thalamocortical VB neurons from wild-type (WT) and leptin-deficient obese (ob/ob) mice...
March 8, 2018: Brain Structure & Function
Rafael Taeho Han, Han-Byul Kim, Young-Beom Kim, Kyungmin Choi, Gi Yeon Park, Pa Reum Lee, JaeHee Lee, Hye Young Kim, Chul-Kyu Park, Youngnam Kang, Seog Bae Oh, Heung Sik Na
Recent studies have provided several lines of evidence that peripheral administration of oxytocin induces analgesia in human and rodents. However, the exact underlying mechanism of analgesia still remains elusive. In the present study, we aimed to identify which receptor could mediate the analgesic effect of intraperitoneal injection of oxytocin and its cellular mechanisms in thermal pain behavior. We found that oxytocin-induced analgesia could be reversed by d(CH2 )5 [Tyr(Me)2 ,Dab5 ] AVP, a vasopressin-1a (V1a) receptor antagonist, but not by desGly-NH2 -d(CH2 )5 [DTyr2 , Thr4 ]OVT, an oxytocin receptor antagonist...
March 2018: Korean Journal of Physiology & Pharmacology
J Wickham, N G Brödjegård, R Vighagen, L H Pinborg, J Bengzon, D P D Woldbye, M Kokaia, M Andersson
Resected hippocampal tissue from patients with drug-resistant epilepsy presents a unique possibility to test novel treatment strategies directly in target tissue. The post-resection time for testing and analysis however is normally limited. Acute tissue slices allow for electrophysiological recordings typically up to 12 hours. To enable longer time to test novel treatment strategies such as, e.g., gene-therapy, we developed a method for keeping acute human brain slices viable over a longer period. Our protocol keeps neurons viable well up to 48 hours...
March 7, 2018: Scientific Reports
Kiyoto Kanbara, Yoshinori Otsuki, Masahito Watanabe, Syunichi Yokoe, Yoshiaki Mori, Michio Asahi, Masashi Neo
BACKGROUND: High-grade chondrosarcoma, which has a high incidence of local recurrence and pulmonary metastasis despite surgical resection, is associated with poor prognosis. Therefore, new and effective adjuvant therapies are urgently required for this disease. Gamma-aminobutyric acid (GABA), which acts as a neurotrophic factor during nervous system development, is related to the proliferation and migration of certain cancer cells. The GABAergic system, which is composed of GABA, the GABA-synthesizing enzyme glutamic acid decarboxylase (GAD), and GABA receptors, has an important function in nerve growth and development of neural crest...
March 7, 2018: BMC Cancer
Marit Poffers, Nathalie Bühne, Christine Herzog, Anja Thorenz, Rongjun Chen, Faikah Güler, Axel Hage, Andreas Leffler, Frank Echtermeyer
BACKGROUND: Voltage-gated sodium channels generate action potentials in excitable cells, but they have also been attributed noncanonical roles in nonexcitable cells. We hypothesize that voltage-gated sodium channels play a functional role during extravasation of neutrophils. METHODS: Expression of voltage-gated sodium channels was analyzed by polymerase chain reaction. Distribution of Nav1.3 was determined by immunofluorescence and flow cytometry in mouse models of ischemic heart and kidney injury...
March 6, 2018: Anesthesiology
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
Qian-Xing Zhuang, Han-Ting Xu, Xu-Juan Lu, Bin Li, Wing-Ho Yung, Jian-Jun Wang, Jing-Ning Zhu
The central histaminergic nervous system, originating from the tuberomammillary nucleus (TMN) of the hypothalamus, widely innervates almost the whole brain, including the basal ganglia. Intriguingly, the histaminergic system is altered in parkinsonian patients. Yet, little is known about the effect and mechanisms of histamine on different types of neurons in the basal ganglia circuitry. Here, by using anterograde tracing, immunostaining, patch clamp recording, and single-cell qPCR techniques, we investigate the histaminergic afferents in the striatum, the major input structure of the basal ganglia, as well as the effect of histamine on the striatal GABAergic medium spiny projection neurons (MSNs)...
March 1, 2018: Molecular Neurobiology
Katharina T Schrapers, Gerhard Sponder, Franziska Liebe, Hendrik Liebe, Friederike Stumpff
Absorption of ammonia from the gastrointestinal tract results in problems that range from hepatic encephalopathy in humans to poor nitrogen efficiency of cattle with consequences for the global climate. Previous studies on epithelia and cells from the native ruminal epithelium suggest functional involvement of the bovine homologue of TRPV3 (bTRPV3) in ruminal NH4+ transport. Since the conductance of TRP channels to NH4+ has never been studied, bTRPV3 was overexpressed in HEK-293 cells and investigated using the patch-clamp technique and intracellular calcium imaging...
2018: PloS One
Jing-Na Ren, Ben-Long Rao, Hong-Xin Ma, Mao-Mao Sha, Yi Kuang, Zheng-Xin Xu
To investigate the effects of isovitexin Ⅳ on transient outward potassium current in rat ventricular myocytes. In this study, MTT assay was used to investigate the safe range of isovitexin. The results showed that the IC₅₀ of the drug was in the range of 10-30 μmol•L⁻¹, and the drug concentration of 1-3 μmol•L⁻¹ for the patch clamp test was within the safe range. In addition, the single ventricular myocytes were obtained by single-enzymatic hydrolysis through aortic retrograde perfusion. The transient outward potassium current (Ito) of rat ventricular myocytes was guided and measured by whole-cell patch-clamp technique and the changes of current characteristics were recorded after isovite was applied...
December 2017: Zhongguo Zhong Yao za Zhi, Zhongguo Zhongyao Zazhi, China Journal of Chinese Materia Medica
Bailin Liu, Yanping Liu, Ruixiu Shi, Xueqin Feng, Xiang Li, Wenna Zhang, Jue Wu, Na Li, Xiuwen Zhou, Miao Sun, Zhice Xu
BACKGROUND/AIMS: Chronic hypoxia in utero could impair vascular functions in the offspring, underlying mechanisms are unclear. This study investigated functional alteration in large-conductance Ca2+-activated K+ (BK) channels in offspring mesenteric arteries following prenatal hypoxia. METHODS: Pregnant rats were exposed to normoxic control (21% O2, Con) or hypoxic (10.5% O2, Hy) conditions from gestational day 5 to 21, their 7-month-old adult male offspring were tested for blood pressure, vascular BK channel functions and expression using patch clamp and wire myograh technique, western blotting, and qRT-PCR...
February 21, 2018: Cellular Physiology and Biochemistry
Nils J G Rorsman, Chau M Ta, Hannah Garnett, Pawel Swietach, Paolo Tammaro
BACKGROUND AND PURPOSE: Optogenetic control of electromechanical coupling in vascular smooth muscle cells (VSMCs) is emerging as a powerful research tool with potential applications in drug discovery and therapeutics. However, the precise ionic mechanisms involved in this control remain unclear. EXPERIMENTAL APPROACH: Cell imaging, patch-clamp electrophysiology, and muscle tension recordings were used to define these mechanisms over a wide range of light stimulations...
February 27, 2018: British Journal of Pharmacology
Fu-Sun Lo, Reha S Erzurumlu
Background: Met receptor tyrosine kinase regulates neurogenesis, differentiation, migration, connectivity, and synaptic plasticity. The human Met gene has been identified as a prominent risk factor for autism spectrum disorder (ASD). Met gene-altered mice serve as useful models for mechanistic studies of ASD. Inactivation of Met in excitatory cortical neurons in mice ( Emx1 cre /Met flox mice) yields a phenotype in which significantly decreased GABAA receptor-mediated inhibition shifts the excitation/inhibition (E/I) balance toward excitation in the somatosensory cortex...
2018: Molecular Autism
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