Isolation and characterization of potential cancer stem cells from solid human tumors--potential applications.
Authors: Dobbin ZC, Landen CN
Abstract
Cancer stem cells (CSCs) are a subpopulation of cells within a heterogeneous tumor that have enhanced biologic properties, e.g., increased capacity for self-renewal, increased tumorigenicity, enhanced differentiation capacity, and resistance to chemo- and radiotherapies. This unit describes protocols to isolate and characterize potential cancer stem cells from a solid tumor. These involve creating a single-cell suspension from tumor tissue, tagging the cell subpopulations of interest, and sorting them into different populations. The sorted subpopulations can be evalua...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Automated Patch Clamp Analysis of nAChα7 and NaV 1.7 Channels.
Authors: Obergrussberger A, Haarmann C, Rinke I, Becker N, Guinot D, Brueggemann A, Stoelzle-Feix S, George M, Fertig N
Abstract
Automated patch clamp devices are now commonly used for studying ion channels. A useful modification of this approach is the replacement of the glass pipet with a thin planar glass layer with a small hole in the middle. Planar patch clamp devices, such as the three described in this unit, are overtaking glass pipets in popularity because they increase throughput, are easier to use, provide for the acquisition of high-quality and information-rich data, and allow for rapid perfusio...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Electrophysiological Studies of Voltage-Gated Sodium Channels Using QPatch HT, an Automated Patch-Clamp System.
Authors: Liu Y
Abstract
Voltage-gated sodium (Nav ) channels are highly sensitive to membrane potential and have fast gating kinetics. Patch clamp electrophysiology has long been the gold standard for studying these channels. Combining high throughput with high information content/accuracy, automated patch clamp technologies have emerged as critical tools in ion channel drug discovery. Described in this unit is the use of QPatch, one of the automated patch clamp systems, to study Nav channel function and pharmacology. Curr. Protoc. Pharmacol. 65:11.14.1-11.14.45. © 2014 by John Wiley & Sons, Inc.
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Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Overview of genetically engineered mouse models of colorectal carcinoma to enable translational biology and drug development.
Authors: Roper J, Martin ES, Hung KE
Abstract
Preclinical models for colorectal cancer (CRC) are critical for translational biology and drug development studies to characterize and treat this condition. Mouse models of human cancer are particularly popular because of their relatively low cost, short life span, and ease of use. Genetically engineered mouse models (GEMMs) of CRC are engineered from germline or somatic modification of critical tumor suppressor genes and/or oncogenes that drive mutations in human disease. Detailed in this overview are the salient features of several useful colorectal cancer GE...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Functional Characterization of Human Stem Cell-Derived Cardiomyocytes.
Authors: Kirsch AG, Obejero-Paz CA, Bruening-Wright A
Abstract
Cardiac toxicity is a leading contributor to late-stage attrition in the drug discovery process and to withdrawal of approved from the market. In vitro assays that enable earlier and more accurate testing for cardiac risk provide early stage predictive indicators that aid in mitigating risk. Human cardiomyocytes, the most relevant subjects for early stage testing, are severely limited in supply. But human stem cell-derived cardiomyocytes (SC-hCM) are readily available from commercial sources and are increasingly used in academic research, drug ...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
In vivo electrophysiological recording techniques for the study of neuropathic pain in rodent models.
Authors: Zhao FY, Jeggo R, Wei H, Whyment A, Fang X, Spanswick D
Abstract
Neuropathic pain develops following nerve injury, and is a chronic pain syndrome that can persist long after repair of a wound or removal of the neurological insult. This condition remains poorly treated, not least because of a lack of mechanism-based therapeutics. Clinically, neuropathic pain is characterized by three major symptoms: thermal or mechanical allodynia (pain sensation in response to previously non-noxious stimuli); hyperalgesia (enhanced pain sensation to noxious stimulation); and spontaneous, ongoing pain. These clinic...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
The DEN and CCl4 -Induced Mouse Model of Fibrosis and Inflammation-Associated Hepatocellular Carcinoma.
Authors: Uehara T, Pogribny IP, Rusyn I
Abstract
Human hepatocellular carcinoma (HCC) develops most often as a complication of fibrosis or cirrhosis. While most human studies of HCC provide crucial insights into the molecular signatures of HCC, seldom do they address the etiology of HCC. Mouse models are essential tools for investigating the pathogenesis of HCC; however, the overwhelming majority of cancer models in rodents do not feature liver fibrosis. Detailed in this unit is a protocol for an experimental mouse model of HCC that arises in association with advanced liver fibrosis. The disease model is i...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Overview of mouse models of autism spectrum disorders.
Authors: Bey AL, Jiang YH
Abstract
This overview describes many well characterized mouse models of autism spectrum disorders (ASDs). Mouse models considered here were selected because they are examples of genetically engineered models where human genetic evidence supports a causative relationship between the targeted mutation and the behavioral phenotype. As the ASD diagnosis is based primarily on behavioral evaluations in humans in the domains of social interaction, communication, and restricted interests, the murine phenotypes analogous to human autistic behaviors are highlighted for the different models...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Overview of pharmacokinetics.
Authors: Aimone LD, de Lannoy IA
Abstract
In addition to having selective potency against the molecular target(s), a compound must be able to reach its intended site of action in vivo in sufficient quantity and for the appropriate duration of time to exert a biological effect. The fate of a compound after in vivo administration depends upon its absorption, distribution, metabolism, and excretion (ADME), as well as its target residence time. The concentration of the compound in the blood, plasma, and other tissues represents the sum of all of these processes. Described in this unit are protocols for adminis...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Modeling human liver cancer heterogeneity: virally induced transgenic models and mouse genetic models of chronic liver inflammation.
Authors: Ringelhan M, Reisinger F, Yuan D, Weber A, Heikenwalder M
Abstract
In addition to being the most common primary liver cancer, hepatocellular carcinoma (HCC) is the second most common cause of cancer-related death in humans. Treatment options are limited for this chemoresistant cancer, with liver transplantation and surgical intervention in early stages being the most successful treatments. Drug development over the past 15 years has focused on generating mouse models that mimic the human pathology for HCC. This has enabled the laboratory testing of potentially new human therapeutics. Described in ...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Overview of different mechanisms of arrestin-mediated signaling.
Authors: Gurevich VV, Gurevich EV
Abstract
Arrestins are characterized by their ability to selectively bind active, phosphorylated GPCRs and suppress (arrest) receptor coupling to G proteins. Nonvisual arrestins are also signaling proteins in their own right, activating a variety of cellular pathways. Arrestins are highly flexible proteins that can assume many distinct conformations. In their receptor-bound conformation, arrestins have higher affinity for a subset of partners. This explains how receptor activation regulates certain branches of arrestin-dependent signaling via arrestin recruitment to GPCRs....
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Arrestin expression in E. coli and purification.
Authors: Vishnivetskiy SA, Zhan X, Chen Q, Iverson TM, Gurevich VV
Abstract
Purified arrestin proteins are necessary for biochemical, biophysical, and crystallographic studies of these versatile regulators of cell signaling. Described herein is a basic protocol for arrestin expression in E. coli and purification of the tag-free wild-type and mutant arrestins. The method includes ammonium sulfate precipitation of arrestins from cell lysates, followed by heparin-Sepharose chromatography. Depending on the arrestin type and/or mutations, the next step is Q-Sepharose or SP-Sepharose chromatography. In many case...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Guinea pig models of asthma.
Authors: McGovern AE, Mazzone SB
Abstract
Described in this unit are methods for establishing guinea pig models of asthma. Sufficient detail is provided to enable investigators to study bronchoconstriction, cough, airway hyperresponsiveness, inflammation, and remodeling. © 2014 by John Wiley & Sons, Inc.
PMID: 25446291 [PubMed - in process] (Source: Current Protocols in Pharmacology)
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Single-channel recording of glutamate receptors.
Authors: Shelley C
Abstract
Highlighted in this unit are issues that should be considered when recording glutamate receptors at the single-channel level, including some commonly encountered problems and their remedies. "UNIT 11.17, Single-Channel Analysis of Glutamate Receptors" describes analysis techniques used to characterize the recorded single-channel properties. © 2015 by John Wiley & Sons, Inc.
PMID: 25737155 [PubMed - in process] (Source: Current Protocols in Pharmacology)
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Single-channel analysis of glutamate receptors.
Authors: Shelley C
Abstract
This is a companion to UNIT 11.16: Single-Channel Recording of Glutamate Receptors. Described here are techniques for analyzing single-channel currents recorded from glutamate receptors to characterize their properties. In addition, issues that need to be taken into account when analyzing glutamate receptor single-channel recording results are discussed. © 2015 by John Wiley & Sons, Inc.
PMID: 25737156 [PubMed - in process] (Source: Current Protocols in Pharmacology)
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
