DOCSLIB.ORG

Role of Voltage-Gated Sodium Channel Isoforms in Electrophysiological Properties of Neurons Innervating the Viscera in Mice

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Role of Voltage-Gated Sodium Channel Isoforms in Electrophysiological Properties of Neurons Innervating the Viscera in Mice Role of Voltage-gated Sodium Channel Isoforms in Electrophysiological Properties of Neurons Innervating the Viscera in Mice Andelain Kristiseter Erickson School of Medicine Discipline of Medicine The University of Adelaide Thesis submitted for the degree of Doctor of Philosophy 17th May 2019 Table of Contents Citation listing ........................................................................................................................................ vi Abstract .................................................................................................................................................. vii Declaration of originality ..................................................................................................................... viii Acknowledgments .................................................................................................................................. ix Abbreviations .......................................................................................................................................... x Chapter 1 Visceral Pain .......................................................................................................................... 2 1.1 Abstract ......................................................................................................................................................... 2 1.2 Introduction .................................................................................................................................................... 2 1.3 Clinical relevance .......................................................................................................................................... 3 1.4 Visceral comorbidities .................................................................................................................................... 3 1.5 Extrinsic sensory afferent innervation of the colon and bladder .................................................................... 4 1.6 Afferent subtypes innervating the colon and bladder .................................................................................... 5 1.7 Conclusions ................................................................................................................................................. 14 1.8 Disclosure statement ................................................................................................................................... 14 1.9 Acknowledgements ..................................................................................................................................... 15 1.10 Literature cited ........................................................................................................................................... 15 Chapter 2 Voltage-gated sodium channels: (Nav)igating the field to determine their contribution to visceral nociception ......................................................................................................................... 24 2.1 Abstract ....................................................................................................................................................... 24 2.2 Introduction .................................................................................................................................................. 25 2.3 Nav1.1 .......................................................................................................................................................... 29 2.4 Nav1.2 .......................................................................................................................................................... 29 2.5 Nav1.3 .......................................................................................................................................................... 29 2.6 Nav1.4 .......................................................................................................................................................... 30 2.7 Nav1.5 .......................................................................................................................................................... 30 2.8 Nav1.6 .......................................................................................................................................................... 30 2.9 Nav1.7 .......................................................................................................................................................... 31 2.10 Nav1.8 ........................................................................................................................................................ 33 2.11 Nav1.9 ........................................................................................................................................................ 34 2.12 Conclusion ................................................................................................................................................. 38 2.13 Additional information ................................................................................................................................ 40 2.14 Author contributions ................................................................................................................................... 40 2.15 Funding ...................................................................................................................................................... 40 2.16 Literature cited ........................................................................................................................................... 40 Chapter 3 Domain organization, gating mechanisms, and neurotoxin binding sites of mammalian voltage-gated sodium channels ........................................................................................................... 48 3.1 Pore-forming α-subunit and auxiliary β-subunits ......................................................................................... 48 3.2 Mechanism of voltage-dependent activation ............................................................................................... 48 ii ` 3.3 Mechanism of fast inactivation ..................................................................................................................... 49 3.4 Nav channel neurotoxin binding sites ........................................................................................................... 49 3.5 Literature cited ............................................................................................................................................. 50 Aims ........................................................................................................................................................ 52 Chapter 4 Methods ................................................................................................................................ 53 4.1 Animals and chronic visceral hypersensitivity model ................................................................................... 53 4.2 Retrograde labeling of colon-innervating neurons and dissociated dorsal root ganglia cell culture ............ 53 4.3 RNA extraction and reverse-transcription quantitative polymerase chain reaction of whole dorsal root ganglia ........................................................................................................................................................................... 54 4.4 Single-cell quantitative polymerase chain reaction of colon-innervating dorsal root ganglia neurons ......... 55 4.5 Whole-cell current-clamp electrophysiology of colon-innervating dorsal root ganglia neurons ................... 55 4.6 Whole-cell voltage-clamp electrophysiology of colon-innervating dorsal root ganglia neurons ................... 56 4.7 Literature cited ............................................................................................................................................. 57 Chapter 5 Active electrophysiological properties of colon-innervating dorsal root ganglia neurons from healthy mice and TNBS-treated mice .......................................................................... 58 5.1 Thoracolumbar and lumbosacral dorsal root ganglia from healthy mice and TNBS-treated mice have similar expression levels of Nav1.1-Nav1.9 (scn1a-scn11a) and β-subunits β1-β4 (scn1b-scn4b) ............................... 58 5.2 Thoracolumbar colon-innervating dorsal root ganglia from healthy mice and TNBS-treated mice have comparable expression of Nav1.1-Nav1.9 (scn1a-scn11a) at the single-cell level ............................................. 60 5.3 Thoracolumbar and lumbosacral colon-innervating dorsal root ganglia from healthy mice express β-subunits β1-β4 (scn1b-scn4b) at the single-cell level ...................................................................................................... 61 5.4 Baseline rheobase in colon-innervating dorsal root ganglia neurons is affected by spinal origin, number of days in culture, soma diameter, and resting membrane potential, but not TNBS treatment .............................. 62 5.5 Colon-innervating dorsal root ganglia neurons from TNBS-treated mice do not display a higher number of action potentials at 2-times rheobase compared to neurons from healthy mice ................................................ 63 5.6 Baseline peak sodium current density in colon-innervating dorsal root ganglia
Load more

Recommended publications
  • Genetic Analysis of the Shaker Gene Complex of Drosophila Melanogaster
    Copyright 0 1990 by the Genetics Society of America Genetic Analysisof the Shaker Gene Complexof Drosophila melanogaster A. Ferriis,* S. LLamazares,* J. L. de la Pornpa,* M. A. Tanouye? and0. Pongs* *Institute Cajal, CSIC, 28006 Madrid, Spain, +California Instituteof Technology, Pasadena, California 91 125, and 'Ruhr Universitat,Bochum, Federal Republic of Germany Manuscript received May 10, 1989 Accepted for publication March 3, 1990 ABSTRACT The Shaker complex (ShC) spans over 350 kb in the 16F region of the X chromosome. It can be dissected by means of aneuploids into three main sections: the maternal effect (ME), the viable (V) and the haplolethal (HL) regions. The mutational analysis of ShC shows a high density of antimorphic mutations among 12 lethal complementation groupsin addition to 14 viable alleles. The complex is the structural locus of a family of potassium channels as well as a number of functions relevant to the biology of the nervous system. The constituents of ShC seem to be linked by functional relationships in view of the similarity of the phenotypes, antimorphic nature of their mutations and the behavior in transheterozygotes. We discuss the relationship between the genetic organization of ShC and the functional coupling of potassium currents with the other functions encodedin the complex. HAKER was the first behavioral mutant detected do not appear tohave the capability of generating, by S in Drosophila melanogaster (CATSCH1944). It was themselves, gated ionic channels. named after another mutantwith a similar phenotype K+ currents are known to be themost diverse class isolated earlier in Drosophila funebris (LUERS 1936). of ionic currents in terms of kinetics, pharmacology The original phenotype was described as the trem- and sensitivity (HILLE 1984; RUDY 1988).Also, these bling of appendagesin the anesthetized fly.
    [Show full text]
  • Expression Profiling of Ion Channel Genes Predicts Clinical Outcome in Breast Cancer
    UCSF UC San Francisco Previously Published Works Title Expression profiling of ion channel genes predicts clinical outcome in breast cancer Permalink https://escholarship.org/uc/item/1zq9j4nw Journal Molecular Cancer, 12(1) ISSN 1476-4598 Authors Ko, Jae-Hong Ko, Eun A Gu, Wanjun et al. Publication Date 2013-09-22 DOI http://dx.doi.org/10.1186/1476-4598-12-106 Peer reviewed eScholarship.org Powered by the California Digital Library University of California Ko et al. Molecular Cancer 2013, 12:106 http://www.molecular-cancer.com/content/12/1/106 RESEARCH Open Access Expression profiling of ion channel genes predicts clinical outcome in breast cancer Jae-Hong Ko1, Eun A Ko2, Wanjun Gu3, Inja Lim1, Hyoweon Bang1* and Tong Zhou4,5* Abstract Background: Ion channels play a critical role in a wide variety of biological processes, including the development of human cancer. However, the overall impact of ion channels on tumorigenicity in breast cancer remains controversial. Methods: We conduct microarray meta-analysis on 280 ion channel genes. We identify candidate ion channels that are implicated in breast cancer based on gene expression profiling. We test the relationship between the expression of ion channel genes and p53 mutation status, ER status, and histological tumor grade in the discovery cohort. A molecular signature consisting of ion channel genes (IC30) is identified by Spearman’s rank correlation test conducted between tumor grade and gene expression. A risk scoring system is developed based on IC30. We test the prognostic power of IC30 in the discovery and seven validation cohorts by both Cox proportional hazard regression and log-rank test.
    [Show full text]
  • Mapping Protein Interactions of Sodium Channel Nav1.7 Using 2 Epitope-Tagged Gene Targeted Mice
    bioRxiv preprint doi: https://doi.org/10.1101/118497; this version posted March 20, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 Mapping protein interactions of sodium channel NaV1.7 using 2 epitope-tagged gene targeted mice 3 Alexandros H. Kanellopoulos1†, Jennifer Koenig1, Honglei Huang2, Martina Pyrski3, 4 Queensta Millet1, Stephane Lolignier1, Toru Morohashi1, Samuel J. Gossage1, Maude 5 Jay1, John Linley1, Georgios Baskozos4, Benedikt Kessler2, James J. Cox1, Frank 6 Zufall3, John N. Wood1* and Jing Zhao1†** 7 1Molecular Nociception Group, WIBR, University College London, Gower Street, 8 London WC1E 6BT, UK. 9 2Mass Spectrometry Laboratory, Target Discovery Institute, University of Oxford, The 10 Old Road Campus, Oxford, OX3 7FZ, UK. 11 3Center for Integrative Physiology and Molecular Medicine, Saarland University, 12 Kirrbergerstrasse, Bldg. 48, 66421 Homburg, Germany. 13 4Division of Bioscience, University College London, Gower Street, London WC1E 6BT, UK.14 15 †These authors contributed equally to directing this work. 16 *Correspondence author. Tel: +44 207 6796 954; E-mail: [email protected] 17 **Corresponding author: Tel: +44 207 6790 959; E-mail: [email protected] 1 bioRxiv preprint doi: https://doi.org/10.1101/118497; this version posted March 20, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 18 Abstract 19 The voltage-gated sodium channel NaV1.7 plays a critical role in pain pathways. 20 Besides action potential propagation, NaV1.7 regulates neurotransmitter release, 21 integrates depolarizing inputs over long periods and regulates transcription.
    [Show full text]
  • Allosteric Features of KCNQ1 Gating Revealed by Alanine Scanning Mutagenesis
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Biophysical Journal Volume 100 February 2011 885–894 885 Allosteric Features of KCNQ1 Gating Revealed by Alanine Scanning Mutagenesis Li-Juan Ma, Iris Ohmert, and Vitya Vardanyan* Institut fu¨r Neurale Signalverarbeitung, Zentrum fu¨r Molekulare Neurobiologie, Universita¨t Hamburg, Hamburg, Germany ABSTRACT Controlled opening and closing of an ion-selective pathway in response to changes of membrane potential is a fundamental feature of voltage-gated ion channels. In recent decades, various details of this process have been revealed with unprecedented precision based on studies of prototypic potassium channels. Though current scientific efforts are focused more on a thorough description of voltage-sensor movement, much less is known about the similarities and differences of the gating mechanisms among potassium channels. Here, we describe the peculiarities of the KCNQ1 gating process in parallel comparison to Shaker. We applied alanine scanning mutagenesis to the S4-S5 linker and pore region and followed the regular- ities of gating perturbations in KCNQ1. We found a fractional constitutive conductance for wild-type KCNQ1. This component increased significantly in mutants with considerably leftward-shifted steady-state activation curves. In contrast to Shaker,no correlation between V1/2 and Z parameters was observed for the voltage-dependent fraction of KCNQ1. Our experimental find- ings are explained by a simple allosteric gating scheme with voltage-driven and voltage-independent transitions. Allosteric features are discussed in the context of extreme gating adaptability of KCNQ1 upon interaction with KCNE b-subunits.
    [Show full text]
  • Engineering Biosynthetic Excitable Tissues from Unexcitable Cells for Electrophysiological and Cell Therapy Studies
    ARTICLE Received 11 Nov 2010 | Accepted 5 Apr 2011 | Published 10 May 2011 DOI: 10.1038/ncomms1302 Engineering biosynthetic excitable tissues from unexcitable cells for electrophysiological and cell therapy studies Robert D. Kirkton1 & Nenad Bursac1 Patch-clamp recordings in single-cell expression systems have been traditionally used to study the function of ion channels. However, this experimental setting does not enable assessment of tissue-level function such as action potential (AP) conduction. Here we introduce a biosynthetic system that permits studies of both channel activity in single cells and electrical conduction in multicellular networks. We convert unexcitable somatic cells into an autonomous source of electrically excitable and conducting cells by stably expressing only three membrane channels. The specific roles that these expressed channels have on AP shape and conduction are revealed by different pharmacological and pacing protocols. Furthermore, we demonstrate that biosynthetic excitable cells and tissues can repair large conduction defects within primary 2- and 3-dimensional cardiac cell cultures. This approach enables novel studies of ion channel function in a reproducible tissue-level setting and may stimulate the development of new cell-based therapies for excitable tissue repair. 1 Department of Biomedical Engineering, Duke University, Durham, North Carolina 27708, USA. Correspondence and requests for materials should be addressed to N.B. (email: [email protected]). NatURE COMMUNicatiONS | 2:300 | DOI: 10.1038/ncomms1302 | www.nature.com/naturecommunications © 2011 Macmillan Publishers Limited. All rights reserved. ARTICLE NatUre cOMMUNicatiONS | DOI: 10.1038/ncomms1302 ll cells express ion channels in their membranes, but cells a b with a significantly polarized membrane that can undergo e 0 a transient all-or-none membrane depolarization (action A 1 potential, AP) are classified as ‘excitable cells’ .
    [Show full text]
  • Cold Sensing by Nav1.8-Positive and Nav1.8-Negative Sensory Neurons
    Cold sensing by NaV1.8-positive and NaV1.8-negative sensory neurons A. P. Luiza, D. I. MacDonalda, S. Santana-Varelaa, Q. Milleta, S. Sikandara, J. N. Wooda,1, and E. C. Emerya,1 aMolecular Nociception Group, Wolfson Institute for Biomedical Research, University College London, London WC1E 6BT, United Kingdom Edited by Peter McNaughton, King’s College London, London, United Kingdom, and accepted by Editorial Board Member David E. Clapham January 8, 2019 (received for review August 23, 2018) The ability to detect environmental cold serves as an important define the distribution and identity of cold-sensitive DRG neurons, survival tool. The sodium channels NaV1.8 and NaV1.9, as well as in live mice, using in vivo imaging. the TRP channel Trpm8, have been shown to contribute to cold sensation in mice. Surprisingly, transcriptional profiling shows that Results NaV1.8/NaV1.9 and Trpm8 are expressed in nonoverlapping neuro- Distribution of DRG Sensory Neurons Responsive to Noxious Cold, in nal populations. Here we have used in vivo GCaMP3 imaging to Vivo. To identify cold-sensitive neurons in vivo, we used a pre- identify cold-sensing populations of sensory neurons in live mice. viously developed in vivo imaging technique to study the responses We find that ∼80% of neurons responsive to cold down to 1 °C do of individual DRG neurons in situ (8). Mice coexpressing Pirt- not express NaV1.8, and that the genetic deletion of NaV1.8 does GCaMP3 (which enables pan-DRG GCaMP3 expression), not affect the relative number, distribution, or maximal response NaV1.8 Cre, and a Cre-dependent reporter (tdTomato) were of cold-sensitive neurons.
    [Show full text]
  • BK Channel Inhibition by Strong Extracellular Acidification
    bioRxiv preprint doi: https://doi.org/10.1101/317339; this version posted May 8, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. BK channel inhibition by strong extracellular acidification Yu Zhou, Xiao-Ming Xia and Christopher J. Lingle Department of Anesthesiology, Washington University School of Medicine, St. Louis, Missouri 63110 Address for correspondence: Yu Zhou Washington University School of Medicine Dept. Anesthesiology, Box 8054 St. Louis MO 63110 email: [email protected] Christopher J. Lingle Washington University School of Medicine Dept. Anesthesiology, Box 8054 St. Louis MO 63110 phone: 314-362-8558 FAX: 314-362-8571 email: [email protected] Running title: BK channel inhibition by extracellular H+ Keywords: extracellular H+, K+ channels, channel inhibition, BK channels, Slo1 channels 1Abbreviations used in this paper: BK, large conductance Ca2+-activated K+ channel 1 bioRxiv preprint doi: https://doi.org/10.1101/317339; this version posted May 8, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. Abstract BK-type voltage- and Ca2+-dependent K+ channels are found in a wide range of cells and intracellular organelles. Among different loci, the composition of the extracellular microenvironment, including pH, may differ substantially. For example, it has been reported that BK channels are expressed in lysosomes with their extracellular side facing the strongly acidified lysosomal lumen (pH ~ 4.5).
    [Show full text]
  • Study of Short Forms of P/Q-Type Voltage-Gated Calcium Channels
    Study of Short Forms of P/Q-Type Voltage-Gated Calcium Channels Qiao Feng Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Graduate School of Arts and Sciences COLUMBIA UNIVERSITY 2017 © 2017 Qiao Feng All rights reserved ABSTRACT Study of Short Forms of P/Q-Type Voltage-Gated Calcium Channels Qiao Feng P/Q-type voltage-gated calcium channels (CaV2.1) are expressed in both central and peripheral nervous systems, where they play a critical role in neurotransmitter release. Mutations in the pore-forming 1 subunit of CaV2.1 can cause neurological disorders such as episodic ataxia type 2, familial hemiplegic migraine type 1 and spinocerebellar ataxia type 6. Interestingly, a 190-kDa fragment of CaV2.1 was found in mouse brain tissue and cultured mouse cortical neurons, but not in heterologous systems expressing full-length CaV2.1. In the brain, the 190-kDa species is the predominant form of CaV2.1, while in cultured cortical neurons the amount of the 190-kDa species is comparable to that of the full-length channel. The 190-kDa fragment contains part of the II-III loop, repeat III, repeat IV and the C-terminal tail. A putative complementary fragment of 80-90 kDa was found along with the 190-kDa form. Moreover, preliminary data show that the abundance of the 190-kDa species and the 80-90-kDa species relative to the full-length channel is upregulated by increased intracellular Ca2+ concentration. Truncation mutations in the P/Q-type calcium channel have been found to cause the neurological disease episodic ataxia type 2.
    [Show full text]
  • (KCND3) (NM 004980) Human Tagged ORF Clone Product Data
    OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for RC222757 Kv4.3 (KCND3) (NM_004980) Human Tagged ORF Clone Product data: Product Type: Expression Plasmids Product Name: Kv4.3 (KCND3) (NM_004980) Human Tagged ORF Clone Tag: Myc-DDK Symbol: KCND3 Synonyms: BRGDA9; KCND3L; KCND3S; KSHIVB; KV4.3; SCA19; SCA22 Vector: pCMV6-Entry (PS100001) E. coli Selection: Kanamycin (25 ug/mL) Cell Selection: Neomycin This product is to be used for laboratory only. Not for diagnostic or therapeutic use. View online » ©2021 OriGene Technologies, Inc., 9620 Medical Center Drive, Ste 200, Rockville, MD 20850, US 1 / 5 Kv4.3 (KCND3) (NM_004980) Human Tagged ORF Clone – RC222757 ORF Nucleotide >RC222757 representing NM_004980 Sequence: Red=Cloning site Blue=ORF Green=Tags(s) TTTTGTAATACGACTCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCC GCCGCGATCGCC ATGGCGGCCGGAGTTGCGGCCTGGCTGCCTTTTGCCCGGGCTGCGGCCATCGGGTGGATGCCGGTGGCCA ACTGCCCCATGCCCCTGGCCCCGGCCGACAAGAACAAGCGGCAGGATGAGCTGATTGTCCTCAACGTGAG TGGGCGGAGGTTCCAGACCTGGAGGACCACGCTGGAGCGCTACCCGGACACCCTGCTGGGCAGCACGGAG AAGGAGTTCTTCTTCAACGAGGACACCAAGGAGTACTTCTTCGACCGGGACCCCGAGGTGTTCCGCTGCG TGCTCAACTTCTACCGCACGGGGAAGCTGCACTACCCGCGCTACGAGTGCATCTCTGCCTACGACGACGA GCTGGCCTTCTACGGCATCCTCCCGGAGATCATCGGGGACTGCTGCTACGAGGAGTACAAGGACCGCAAG AGGGAGAACGCCGAGCGGCTCATGGACGACAACGACTCGGAGAACAACCAGGAGTCCATGCCCTCGCTCA GCTTCCGCCAGACCATGTGGCGGGCCTTCGAGAACCCCCACACCAGCACGCTGGCCCTGGTCTTCTACTA
    [Show full text]
  • The Nav1.7 Sodium Channel: from Molecule to Man
    REVIEWS The NaV1.7 sodium channel: from molecule to man Sulayman D. Dib-Hajj1,2, Yang Yang1,2, Joel A. Black1,2 and Stephen G. Waxman1,2 Abstract | The voltage-gated sodium channel NaV1.7 is preferentially expressed in peripheral somatic and visceral sensory neurons, olfactory sensory neurons and sympathetic ganglion neurons. NaV1.7 accumulates at nerve fibre endings and amplifies small subthreshold depolarizations, poising it to act as a threshold channel that regulates excitability. Genetic and functional studies have added to the evidence that NaV1.7 is a major contributor to pain signalling in humans, and homology modelling based on crystal structures of ion channels suggests an atomic-level structural basis for the altered gating of mutant NaV1.7 that causes pain. Neuropathic pain Voltage-gated sodium channels are essential for electro- that are not seen when these channels are expressed in 11 Pain resulting from lesions or genesis in excitable cells. Nine pore-forming α‑subunits HEK 293 cells . Methods are now available that allow the diseases of the somatosensory of such channels (referred to as channels hereinafter), expression and functional profiling of sodium channels in system. 1 NaV1.1–NaV1.9, have been identified in mammals . peripheral neurons, which more closely mimic the in vivo These isoforms share a common overall structural environment of such channels12. motif (FIG. 1). They are each composed of a long poly- In humans, gain‑of‑function mutations in SCN9A, neuropathic pain peptide (1,700–2,000 amino acids) that folds into four which encodes NaV1.7, lead to severe , homologous domains (DI–DIV) that are linked by three whereas loss‑of‑function mutations in this gene lead to loops (L1–L3), with each domain having six transmem- an indifference to pain13.
    [Show full text]
  • Roderick Mackinnon
    POTASSIUM CHANNELS AND THE ATOMIC BASIS OF SELECTIVE ION CONDUCTION Nobel Lecture, December 8, 2003 by Roderick MacKinnon Howard Hughes Medical Institute, Laboratory of Molecular Neurobiology and Biophysics, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. INTRODUCTION All living cells are surrounded by a thin, approximately 40 Å thick lipid bilay- er called the cell membrane. The cell membrane holds the contents of a cell in one place so that the chemistry of life can occur, but it is a barrier to the movement of certain essential ingredients including the ions Na+, K+, Ca2+ and Cl-. The barrier to ion flow across the membrane – known as the dielec- tric barrier – can be understood at an intuitive level: the cell membrane inte- rior is an oily substance and ions are more stable in water than in oil. The en- ergetic preference of an ion for water arises from the electric field around the ion and its interaction with neighboring molecules. Water is an electrically polarizable substance, which means that its molecules rearrange in an ion’s electric field, pointing negative oxygen atoms in the direction of cations and positive hydrogen atoms toward anions. These electrically stabilizing interac- tions are much weaker in a less polarizable substance such as oil. Thus, an ion will tend to stay in the water on either side of a cell membrane rather than en- ter and cross the membrane. And yet numerous cellular processes, ranging from electrolyte transport across epithelia to electrical signal production in neurons, depend on the flow of ions across the membrane.
    [Show full text]
  • Anti-KCNB1 / Kv2.1 Antibody (ARG59672)
    Product datasheet [email protected] ARG59672 Package: 50 μg anti-KCNB1 / Kv2.1 antibody Store at: -20°C Summary Product Description Rabbit Polyclonal antibody recognizes KCNB1 / Kv2.1 Tested Reactivity Hu, Ms, Rat Tested Application IHC-Fr, IHC-P, WB Host Rabbit Clonality Polyclonal Isotype IgG Target Name KCNB1 / Kv2.1 Antigen Species Human Immunogen Recombinant protein corresponding to V687-I858 of Human Kv2.1. Conjugation Un-conjugated Alternate Names Voltage-gated potassium channel subunit Kv2.1; DRK1; EIEE26; Delayed rectifier potassium channel 1; h- DRK1; Potassium voltage-gated channel subfamily B member 1; KV2.1 Application Instructions Application table Application Dilution IHC-Fr 0.5 - 1 µg/ml IHC-P 0.5 - 1 µg/ml WB 0.1 - 0.5 µg/ml Application Note IHC-P: Antigen Retrieval: By heat mediation. * The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist. Calculated Mw 96 kDa Properties Form Liquid Purification Affinity purification with immunogen. Buffer 0.9% NaCl, 0.2% Na2HPO4, 0.05% Sodium azide and 5% BSA. Preservative 0.05% Sodium azide Stabilizer 5% BSA Concentration 0.5 mg/ml Storage instruction For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C or below. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles. Suggest spin the vial prior to opening. The antibody solution should be gently mixed www.arigobio.com 1/4 before use. Note For laboratory research only, not for drug, diagnostic or other use.
    [Show full text]