Ion Channel Factsbook: Extracellular Ligand-Gated Channels

By Academic Press

How do you keep track of basic information on the proteins you work with? Where do you find details of their physicochemical properties, sequence information, gene organization? Are you tired of scanning review articles, primary papers and databases to locate that elusive fact?

The Academic Press FactsBook series will satisfy scientists and clinical researchers suffering from information overload. Each volume provides a catalogue of the essential properties of families of molecules. Gene organization, sequence information, physicochemical properties, and biological activity are presented using a common, easy to follow format. Taken together they compile everything you wanted to know about proteins but were too busy to look for.In a set of four inter-related volumes, The Ion Channel FactsBook provides a comprehensive framework of facts about channel molecules central to electrical signalling phenomena in living cells.

The first volume is devoted to Extracellular Ligand-Gated Integral Receptor-Channel Families including those molecular complexes activated by: 5-Hydroxytryptamine, ATP, Glutamate, Acetylcholine, GABA, Glycine.

• Nomenclature
• Expression
• Sequence analyses
• Structure and function
• Electrophysiology
• Pharmacology
• Information retrieval

• Language: English
• Publisher: Academic Press
• Release date: Nov 16, 1995
• ISBN: 9780080535197

Book preview

THE ION CHANNEL FactsBook

Extracellular Ligand-Gated Channels

First Edition

Edward C. Conley

Molecular Pathology, c/o Ion Channel/Gene Expression University of Leicester/Medical Research Council Centre for Mechanisms of Human Toxicity, UK

with contributions from

William J. Brammar

Department of Biochemistry,

University of Leicester, UK

Academic Press

Harcourt Brace & Company, Publishers

LONDON SAN DIEGO NEW YORK BOSTON

SYDNEY TOKYO TORONTO

Table of Contents

Cover image

Inside Front Cover

Title page

Copyright page

Cumulative table of contents for Volumes I to IV

Acknowledgements

Entry 02: Introduction & layout of entries

HOW TO USE THE ION CHANNEL FACTSBOOK

GUIDE TO THE PLACEMENT CRITERIA FOR EACH FIELD

Abbreviations

Entry 04: ELG Key facts: Extracellular ligand-gated receptor–channels – key facts

Entry 05: ELG CAT 5-HT3: Extracellular 5-hydroxytrypt- amine-gated integral receptor–channels

NOMENCLATURES

EXPRESSION

SEQUENCE ANALYSES

STRUCTURE & FUNCTIONS

ELECTROPHYSIOLOGY

PHARMACOLOGY

INFORMATION RETRIEVAL

Entry 06: ELG CAT ATP: Extracellular ATP-gated receptor-channels (P2XR)

NOMENCLATURES

EXPRESSION

SEQUENCE ANALYSES

STRUCTURE & FUNCTIONS

ELECTROPHYSIOLOGY

PHARMACOLOGY

INFORMATION RETRIEVAL

Entry 07: ELG CAT GLU AMPA/KAIN: AMPA/kainate-selective (non-NMDA) glutamate receptor–channels

NOMENCLATURES

EXPRESSION

SEQUENCE ANALYSES

STRUCTURE & FUNCTIONS

ELECTROPHYSIOLOGY

PHARMACOLOGY

INFORMATION RETRIEVAL

Entry 08: ELG CAT GLU NMDA: N-Methyl-D-aspartate (NMDA)-selective glutamate receptor–channels

NOMENCLATURES

EXPRESSION

mRNA distribution

SEQUENCE ANALYSES

STRUCTURE & FUNCTIONS

ELECTROPHYSIOLOGY

PHARMACOLOGY

INFORMATION RETRIEVAL

Entry 09: ELG CAT nAChR: Nicotinic acetylcholine-gated integral receptor–channels

NOMENCLATURES

EXPRESSION

SEQUENCE ANALYSES

STRUCTURE & FUNCTIONS

ELECTROPHYSIOLOGY

PHARMACOLOGY

INFORMATION RETRIEVAL

Entry 10: ELG C1 GABAA: Inhibitory receptor-channels gated by extracellular gamma-aminobutyric acid

NOMENCLATURES

EXPRESSION

SEQUENCE ANALYSES

STRUCTURE & FUNCTIONS

ELECTROPHYSIOLOGY

PHARMACOLOGY

INFORMATION RETRIEVAL

Entry 11: ELG C1 GLY: Inhibitory receptor–channels gated by glycine

NOMENCLATURES

EXPRESSION

SEQUENCE ANALYSES

STRUCTURE &. FUNCTIONS

ELECTROPHYSIOLOGY

PHARMACOLOGY

INFORMATIOIN RETRIEVAL

Entry 12: Feedback and access to the Cell-Signalling Network (CSN)

FEEDBACK

GUIDELINES ON THE TYPES OF FEEDBACK REQUIRED

THE CELL-SIGNALLING NETWORK (from-Mid-l996)

Entry 13: Entry Number Rubric

Index

Inside Front Cover

Other books in the FactsBook Series:

A. Neil Barclay, Albertus D. Beyers, Marian L. Birkeland, Marion H. Brown, Simon J. Davis, Chamorro Somoza and Alan F. Williams

The Leucocyte Antigen FactsBook

Robin Callard and Andy Gearing

The Cytokine FactsBook

Steve Watson and Steve Arkinstall

The G-Protein Linked Receptor FactsBook

Rod Pigott and Christine Power

The Adhesion Molecule FactsBook

Shirley Ayad, Ray Boot-Handford, Martin J. Humphries, Karl E. Kadler and C. Adrian Shuttleworth

The Extracellular Matrix FactsBook

Robin Hesketh

The Oncogene FactsBook

Grahame Hardie and Steven Hanks

The Protein Kinase Factsbook

Copyright

This book is printed on acid-free paper

ACADEMIC PRESS LIMITED

24–28 Oval Road

LONDON NW1 7DX

United States Edition Published by

ACADEMIC PRESS INC.

San Diego, CA 92101

Copyright © 1996 by

ACADEMIC PRESS LIMITED

All rights reserved

No part of this book may be reproduced in any form by photostat, microfilm, or by any other means, without written permission from the publishers

A catalogue record for this book is available from the British Library

ISBN 0-12-184450-1

Typeset by Alden Multimedia, Oxford and Northampton

Printed and bound in Great Britain by

WBC, Bridgend, Mid Glam.

Cumulative table of contents for Volumes I to IV

Contents

Cumulative table of contents for Volumes I to IV (entry 01)

Acknowledgements

Introduction and layout of entries (entry 02)

How to use The Ion Channel FactsBook

Guide to the placement criteria for each field

Abbreviations (entry 03)

VOLUME I

EXTRACELLULAR LIGAND-GATED CHANNELS

VOLUME II

INTRACELLULAR LIGAND-GATED CHANNELS

VOLUME III

INWARD RECTIFIER AND INTERCELLULAR CHANNELS

VOLUME IV

VOLTAGE-GATED CHANNELS

ION CHANNEL RESOURCES

Feedback

Comments and suggestions regarding the scope, arrangement and other matters relating to the coverage/contents can be sent to the e-mail feedback file [e-mail]. (see field 57 of most entries for further details)

Acknowledgements

Thanks are due to the following people for their time and help during compilation of the manuscripts: Professors Peter Stanfield, Nick Standen and Gordon Roberts (Leicester), and Ole Petersen (Liverpool) for advice, to Allan Winter, Angela Baxter, Shelly Hundal, Phil Shelton and Sue Robinson for help with photocopying, to Chris Hankins and Richard Mobbs of the Leicester University Computer Centre, and to Dr Tessa Picknett and Chris Gibson of Academic Press for their enthusiasm and patience.

Gratitude is also expressed to all of the anonymous manuscript readers who supplied much constructive feedback, as well as the following who provided advice, information and encouragement: Stephen Ashcroft (Oxford), Eric Barnard (London), Dale Benos (Harvard), William Catterall (Washington), K. George Chandy (UC Irvine), Peter Cobbold (Liverpool), David Clapham (Mayo Foundation), Noel Davies (Leicester), Dario DiFrancesco (Milano), Ian Forsythe (Leicester), Sidney Fleischer (Vanderbilt), George Gutman (UC Irvine), Richard Haugland (Molecular Probes, Inc.), Bertil Hille (Washington), Michael Hollmann (G6ttingen), Anthony Hope (Dundee), Benjamin Kaupp [Iulich], Jeremy Lambert (Dundee), Shigetada Nakanishi (Kyoto), Alan North (Glaxo Institute for Molecular Biology), John Peters (Dundee), Olaf Pongs (Hamburg), David Spray (Yeshiva), Kent Springer (Institute for Scientific Information), Steve Watson (Oxford), Paul Van Houlte (I.R.I.S.) and Steven Wertheim (Harvard).

Thanks are also due to the Department of Pathology at the University of Leicester, Harcourt Brace, the Medical Research Council and Zeneca Pharma-ceuticals, for providing generous sponsorship, equipment and facilities.

We would like to acknowledge the authors of all those papers and reviews which in the interest of completeness we have quoted, but have not had space to cite directly.

ECC would like to thank Professors Denis Noble in Oxford and Anthony Campbell in Cardiff, Tony Buzan in Winton, Dorset and Richard Gregory in Bristol for help and inspiration, and would like to dedicate his contributions to Paula, Rebecca and Katharine for all their love and support over the past four years.

Left: Edward Conley, Right: William Brammer

Entry 02

Introduction & layout of entries

Edward C. Conley

The Ion Channels FactsBook is intended to provide a 'summary of molecular properties' for all known types of ion channel protein in a cross-referenced and 'computer-updatable' format. Today, the subject of ion channel biology is an extraordinarily complex one, linking several disciplines and technologies, each adding its own contribution to the knowledge base. This diversity of approaches has left a need for accessible information sources, especially for those reading outside their own field. By presenting 'facts' within a systematic framework, the FactsBook aims to provide a 'logical place to look' for specific information when the need arises. For students and researchers entering the field, the weight of the existing literature, and the rate of new discoveries, makes it difficult to gain an overview. For these readers, The Ion Channels FactsBook is written as a directory, designed to identify similarities and differences between ion channel types, while being able to accommodate new types of data within the framework. The main advantages of a systematic format is that it can speed up identification of functional links between any 'facts' already in the database and maybe provide a raison d’être for specific experiments where information is not known. Although such 'facts' may not go out-of-date, interpretations based on them may change considerably in the light of additional, more direct evidence. This is particularly true for the explosion of new information that is occurring as a direct consequence of the molecular cloning of ion channel genes. It can be anticipated that many more ion channel genes will be cloned in the near future, and it is also likely that their functional diversity will continue to exceed expectations based on pharmacological or physiological criteria alone.

An emphasis on properties emergent from ion channel molecular functions

Understanding how the interplay of currents through many specific ion channel molecules determines complex electrophysiological behaviour of cells remains a significant scientific challenge. The approach of the FactsBook is to associate and relate this complex cell phenotypic behaviour (e.g. its physiology and pharmacology) to ion channel gene expression-control wherever possible even where the specific gene has not yet been cloned. Thus the ion channel molecule becomes the central organizer, and accordingly arbitrates whether information or topics are included, emphasized, sketched-over or excluded. In keeping with this, ion channel characteristics are described in relation to known structural or genetic features wherever possible (or where they are ultimately molecular characteristics). Invariably, this relies on the availability of sequence data for a given channel or group of channels. However, a number of channel types exist which have not yet been sequenced, or display characteristics in the native form which are not precisely matched by existing clones expressed in heterologous cells (or are otherwise ambiguously classified). To accommodate these channel types, summaries of characteristics are included in the standard entry field format, with inappropriate fieldnames omitted. Thus the present 'working arrangement' of entries and fields is broad enough to include both the 'cloned' and 'uncloned' channel types, but in due course will be gradually supplanted by a comprehensive classification based on gene locus, structure, and relatedness of primary sequences. In all cases, the scope of the FactsBook entries is limited to those proteins forming (or predicted to form) membrane-bound, integral ionic channels by folding and association of their primary protein sequences. Activation or suppression of the channel current by a specified ligand or voltage step is generally included as part of the channel description or name (see below). Thus an emphasis is made throughout the book on intrinsic features of channel molecule itself and not on those of separately encoded, co-expressed proteins. In the present edition, there is a bias towards descriptions of vertebrate ion channels as they express the full range of channel types which resemble characteristics found in most eukaryotes.

Anticipated development of the dataset – Integration of functional information around molecular types

Further understanding of complex cellular electrical and pharmacological behaviour will not come from a mere catalogue of protein properties alone. This book therefore begins a process of specific cross-referencing of molecular properties within a functional framework. This process can be extended to the interrelationships of ion channels and other classes of cell-signalling molecules and their functional properties. Retaining protein molecules (i.e. gene products) as 'fundamental units of classification' should also provide a framework for understanding complex physiological behaviour resulting from co-expressed sets of proteins. Significantly, many pathophysiological phenotypes can also be linked to selective molecular 'dysfunction' within this type of framework. Finally, the anticipated growth of raw sequence information from the human genome project may reveal hitherto unexpected classes and subtypes of cell-signalling components - in this case the task then will be to integrate these into what is already known (see also description of Field number 06: Subtype classifications and Field number 05: Gene family).

The Cell-Signalling Network (CSN)

From the foregoing discussion, it can be seen that establishment and consolidation of an integrated 'consensus database' for the many diverse classes of cell signalling molecules (including, for example, receptors, G proteins, ion channels, ion pumps, etc.) remains a worthwhile goal. Such a resource would provide a focus for identifying unresolved issues and may avoid unnecessary duplication of research effort. Work has begun on a prototype cell-signalling molecule database cooperatively maintained and supported by contributions from specialist groups world-wide: The Cell-Signalling Network (CSN) operating from mid-1996 under the World Wide Web† of the Internet† has been designed to disseminate consensus properties of a wide range of molecules involved in cell signal transduction. While it may take some time (and much good-will) to establish a comprehensive network, the many advantages of such a co-operative structure are already apparent. Immediately, these include an 'open' mechanism for consolidation and verification of the dataset, so that it holds a 'consensus' or 'validated' set of information about what is known about each molecule and practical considerations such as nomenclature recommendations (see, for example, the IUPHAR nomenclature sections under the CSN 'home page').

The CSN also allows unlimited cross-referencing by pointing to related information sets, even where these are held in multiple centres around the world. On-line support for technical terms (glossary items, indicated by dagger symbols (†) throughout the text) and reference to explanatory appendices (e.g. on associated signalling components such as G protein†-linked receptors†) are already supported for use with this book. Eventually, benefits could include (for instance) direct 'look-up' of graphical resources for protein structure, in situ and developmental gene expression atlases†, interactive molecular models for structure/function analysis, DNA/protein sequences linked to feature tables, gene mapping resources and other pictorial data. These developments (not all are presently supported) will use interactive electronic media for efficient browsing and maintenance. For a brief account of the Cell-Signalling Network, see Feedback & CSN access, entry 12. For a full specification, see Resource J – Search criteria & CSN development, entry 65.

HOW TO USE THE ION CHANNEL FACTSBOOK

Common formats within the entries

A proposed organizational hierarchy for information about ion channel molecules

Information on named channel types is grouped in entries under common headings which repeat in a fixed order - e.g. for ion channel molecules which have been sequenced, there are broad sections entitled NOMENCLATURES, EXPRESSION, SEQUENCE ANALYSES, STRUCTURE & FUNCTIONS, ELECTROPHYSIOLOGY, PHARMACOLOGY, INFORMATION RETRIEVAL and REFERENCES, in that order. Within each section, related fieldnames are listed, always in alphabetical order and indexed by a field number (see below), which makes electronic cross-referencing and 'manual' comparisons easier.

While the sections and fields are not rigid categories, an attempt has been made to remain consistent, so that corresponding information for two different channels can be looked up and compared directly. If a field does not appear, either the information was not known or was not found during the compilation period. Pertinent information which has been published but is absent from entries would be gratefully received and will be added to the 'entry updates' sections within the CSN (see Feedback & CSN access, entry 12). Establishment of this 'field' format has been designed so that every available 'fact' should have its logical 'place'. In the future, this arrangement may help to establish 'universally accepted' or 'consensus' properties of any given ion channel or other cell-signalling molecule. This validation process critically depends on user feedback to contributing authors. The CSN (above) establishes an efficient electronic mechanism to do this, for continual refinement of entry contents.

Independent presentation of 'facts' and conventions for cross-referencing

The FactsBook departs from a traditional review format by presenting its information in related groups, each under a broader heading. Entries are not designed or intended to be read 'from beginning to end', but each 'fact' is presented independently under the most pertinent fieldname. Independent citation of 'facts' may sometimes result in some repetition (redundancy) of general principles between fields, but if this is the case some effort has been made to 'rephrase' these for clarity (suggested improvements for presentation of any 'fact' are welcome - see Field number 57: Feedback).

For readers unfamiliar with the more general aspects of ion channel biology, some introductory information applicable to whole groups of ion channel molecules is needed, and this is incorporated into the 'key facts' sections preceding the relevant set of entries. These sections, coupled with the 'electronically updated' glossary items (available on-line, and indicated by the dagger† symbol, see below) provide a basic overview of principles associated with detailed information in the main entries of the book.

Extensive cross-referencing is a feature of the book. For example, cross-references between fields of the same entry are of the format (see Fieldname, xx-yy). Cross-references between fields of different channel type entries are generally of the format see fieldname under SORTCODE, xx-yy; for example - see mRNA distribution, under ELG Cl GABAA' 10-13. This alphabetical 'sortcode' and numerical 'entry numbers' (printed in the header to each page) are simply devices to make cross-referencing more compact and to arrange the entries in an approximate running order based on physiological features such as mode of gating†, ionic selectivity†, and agonist† specificity. A 'sort order' based on physiological features was judged to be more intuitive for a wider readership than one based on gene structure alone, and enables 'cloned' and 'uncloned' ion channel types to be listed together. The use and criteria for sortcode designations are described under the subheading Derivation of the sortcode (see Field number 02: Category (sortcode)). Entry 'running order' is mainly of importance in book-form publications. New entries (or mergers/subdivisions between existing entries) will use serial entry numbers as 'electronic pointers' to appropriate files.

Cross-references are frequently made to an on-line index of glossary items by dagger symbols† wherever they might assist someone with technical terms and concepts when reading outside their own field. The glossary is designed to be used side-by-side with the FactsBook entries and is accessible in updated form over the Internet†/World Wide Web† with suitable browsing† software (for details, see Feedback & CSN access, entry 12).

Contextual markers and styles employed within the entries

Throughout the books, a six-figure index number (xx-yy-zz, e.g. 19-44-01: ) separates groups of facts about different aspects of the channel molecule, and carries information about channel type/entry number (e.g. 19- ∼ InsP3 receptor-channels), information type/field number (e.g. -44-, Channel modulation) and running paragraph number (datatype) (e.g. -01). This simple 'punctate' style has been adopted for maximum flexibility of updating (both error-correction and consolidation with new information), cross-referencing and multi-authoring. The CSN specification (see entry 65) includes longer term plans to structure field-based information into convenient data-types which will be indexed by a zz numerical designation.

Italicized subheadings are employed to organize the facts into related topics where a field has a lot of information associated with it. Specific illustrated points or features within a field are referenced to adjacent figures. Usage of abbreviations and common symbols are defined in context and/or within the main abbreviations index at the front of each book. Abbreviated chemical names and those of proprietary pharmaceutical compounds are listed within the electronically updated Resource C – Compounds & proteins, also available via the 'home page' of the Cell-Signalling Network.

Generally, highlighting of related subtopics emergent from the molecular properties ('facts') associated with the ion channel under description are indicated within a field by lettering in bold. All subtopics are cross-referenced by means of a large cumulative subject index (entry 66), which can permit retrieval of information by topic without requiring prior knowledge of ion channel properties. Throughout the main text, italics draw attention to special cases, caveats, hypotheses and exceptions. The 'Note:' prefix has been used to indicate supplemental or comparative information of significance to the quoted data in context.

Special considerations for integrating properties derived from 'cloned' and 'native' channels

While a certain amount of introductory material is given to set the context, the emphasis on molecular properties means the treatment of many important biological processes or phenomena is reduced to a bare outline. References given in the Related sources and reviews field and the electronically updated Resource F – Supplementary ion channel reviews accessible via the CSN (see Feedback & CSN access, entry 12) are intended to address this imbalance.

For summaries of key molecular features, a central channel 'protein domain topography model' is presented. Individual features that are illustrated on the protein domain topography model are identified within the text by the symbol [PDTM].

Wherever molecular subtype-specific data are quoted (such as the particular behaviour of a ion channel gene family† member or isoform†) a convention of using the underlined trivial or systematic name as a prefix has been adopted – e.g. mIRKl:; RCK1:; Kv3.1: etc.

GUIDE TO THE PLACEMENT CRITERIA FOR EACH FIELD

Criteria for NOMENCLATURES sections

This section should bring together for comparison present and previous names of ion channels or currents, with brief distinctions between similar terms. Where systematic names have already been suggested or adopted by published convention, they should be included and used in parallel to trivial names.

Field number 01: Abstract/general description

This field should provide a summary of the most important functional characteristics associated with the channel type.

Field number 02: Category (sortcode)

The alphabetical 'sortcode' should be used for providing a logical running order for the individual entries which make up the book. It is not intended to be a rigorous channel classification, which is under discussion, but rather a practical index for finding and cross-referencing information, in conjunction with the six-figure index number (see above). The Category (sortcode) field also lists a designated electronic retrieval code (unique embedded identifier or UEI) for 'tagging' of new articles of relevance to the contents of the entry. For further details on the use and implementation of UEIs, see the description for Resource J (in this entry) and for a full description, see Resource J – Search criteria & CSN development, entry 65.

Derivation of the sortcode:

Although we do not yet have a complete knowledge of all ion channel primary† structures, knowledge of ion channel gene family† and superfamily† structure allows a working sort order to be established. To take an example, the extracellular ligand-gated (ELG) receptor-channels share many structural features, which reflects the likely duplication and divergent evolution of an ancestral gene. The present-day forms of such channels reflect the changes that have occurred through adaptive radiation† of the ancestral type, particularly for gating† mechanism and ionic selectivity† determinants. Thus, the entry running order (alphabetical, via the sortcode) of the FactsBook entries should depend primarily on these two features. The sortcode therefore consists of several groups of letters, each denoting a characteristic of the channel molecule: Entries are sorted first on the principal means for channel gating† (first three letters), whether this is by an extracellular ligand† (ELG), small intracellular ligand† (ILG) or transmembrane voltage (VLG). For convenience, the ILG entries also include certain channels which are obligately dependent on both ligand binding and hydrolysis for their activation - e.g. channels of the ATP-binding cassette (ABC) superfamily. Other channel types may he subject to direct mechanical gating (MEC) or sensitive to changes in osmolarity (OSM) – see the Cumulative tables of contents and the first page of each entry for descriptions and scope. Due to their unusual gating characteristics, a separate category (INR) has been created for inward rectifier-type channels.

The second sort (the next three letters of the sortcode) should be on the basis of the principal permeant ions, and may therefore indicate high selectivity for single ions (e.g. Ca, Cl, K, Na] or multiple ions of a specified charge (e.g. cations – CAT). Indefinite sortcode extensions can be assigned to the sortcode if it is necessary to distinguish similar but separately encoded groups of channels (e.g. compare ELG C1 GABAA' entry 10 and ELG C1 (GLY, entry 11).

Field number 03: Channel designation

This field should contain a shorthand designation for the ion channel molecule - mostly of the form XY or X[Y] where X denotes the major ionic permeabilities† (e.g. K, Ca, cation) and Y denotes the principal mechanism of gating†where this acts directly on the channel molecule itself (e.g. cGMP, voltage, calcium, etc.). Otherwise, this field contains a shorthand designation for the channel which is used in the entry itself.

Field number 04: Current designation

This field should contain a shorthand designation for ionic currents conducted by the channel molecule, which is mostly of the form IX[Y], IX,Y or IX-Y where X and Y are defined as above.

Field number 05: Gene family

This field should indicate the known molecular relationships to other ion channels or groups of ion channels at the level of amino acid primary sequence homology†, within gene families† or gene superfamilies†. Where multiple channel subunits are encoded by separate genes, a summary of their principal features should be tabulated for comparison. Where the gene family is particularly large, or cannot be easily described by functional variation, a gene family tree† derived by a primary sequence alignment algorithm† (see Resource D – 'Diagnostic' tests, entry 59) may be included as a figure in this field.

Field number 06: Subtype classifications

This field should include supplementary information about any schemes of classification that have been suggested in the literature. Generally, the most robust schemes are those based on complete knowledge of gene family† relationships (see above) and this method can identify similarities that are not easily discernible by pharmacological or electrophysiological criteria alone - see, for example, the entries JUN (connexins), entry 35, and INR K (subunits), entry 33. Note, however, that some native† channel types are more conveniently 'classified' by functional or cell-type expression parameters which take into account interactions of channels with other co-expressed proteins (see, for example, discussion pertaining to the cyclic nucleotide-gated (CNG-) channel family in the entries ILG Key facts, entry 14, ILG CAT cAMP, entry 21, and ILG CAT cGMP, entry 22. Debate on the 'best' or 'most appropriate' channel classification schemes is likely to continue for some time, and it is reasonable to suppose that alternative subtype classifications may be applied and used by different workers for different purposes.

Since the 'running order' of the FactsBook categories depends on inherent molecular properties of channel cDNAs†, genes† or the expressed proteins, future editions will gradually move to classification on the basis of separable gene loci†. Thus multiple channel protein variants resulting from processes of alternative RNA splicing† but encoded by a single gene locus† will only ever warrant one 'channel-type' entry (e.g. see BKCa variants under ILG K Ca, entry 27). Distinct proteins resulting from transcription† of separable gene loci, for example in the case of different gene family members, will (ultimately) warrant separate entries. For the time being, there is insufficient knowledge about the precise phenotypic† roles of many 'separable' gene family members to justify separate entries (as in the case of the VLG K KV series entries).

Classification by gene locus designation (see Field number 18: Chromosomal location) can encompass all structural and functional variation, while being 'compatible' with efforts directed to identifying phenotypic and pathophysiological† roles of individual gene products (e.g. by gene-knockout†, locus replacement† or disease-linked gene mapping† procedures - see Resource D – 'Diagnostic' tests, entry 59). Subtype classifications based on gene locus control can also incorporate the marked developmental changes which pertain to many ion channel genes (see Field number 11: Developmental regulation) and can be implemented when the 'logic' underlying gene expression-control† for each family member is fully appreciated. A 'genome-based' classification of FactsBook entries may also help comprehend and integrate equivalent information based for other ('non-channel') cell-signalling molecules (see Resources G, H and I, entries 62, 63 and 64).

Field number 07: Trivial names

This field should list commonly used names for the ion channel (or its conductance†). Often a channel will be (unsystematically) named by its tissue location or unusual pharmacological/physiological properties, and these are also listed in this field. While unsystematic names do not indicate molecular relatedness, they are often more useful for comparative/descriptive purposes. For these and historical reasons, trivial names (e.g. clone/isolate names for K+ channel isoforms) are used side-by-side with systematic names, where these exist. A standardized nomenclature for ion channels is under discussion, e.g. see the series of articles by Pongs, Edwards, Weston, Chandy, Gutman, Spedding and Vanhoutte in Trends Pharmacol Sci (1993) 14: 433–6. Future recommendations on standardized nomenclature will appear in files accessible under the IUPHAR entry of the Cell-Signalling Network (see Feedback & CSN access, entry 12).

Criteria for EXPRESSION sections

This section should bring together information on expression patterns of the ion channel gene, indicating functional roles of specific channels in the cell type or organism. The complex and profound roles of ionic currents in vertebrate development (linking plasma membrane signalling and genome activation) are also emphasized within the fields of this section.

Field number 08: Cell-type expression index

Comprehensive systems relating the expression of specified molecular components to specified anatomical and developmental loci ('expression atlases') are being developed in a number of centres and in due course will form a superior organizational framework for this type information (see discussion below). In the meantime, the range of cell-type expression should be indicated in this field in the form of alphabetized listings. Notably, there is a substantial literature concerned with the electrophysiology of ion channels where the tissue or cell type forms the main focus of the work. In some cases, this has resulted in detailed 'expression surveys', revealing properties of interacting sets of ion channels, pumps, transporters and associated receptors. Such review-type information is of importance when discussing the contribution of individual ion channel molecules to a complex electrophysiological phenotype† and/or overall function of the cell. For further references to 'cell-type-selective' reviews, see Resource H - Listings of cell types, entry 63 accessible via the CSN (see Feedback & CSN access, entry 12).

Problems and opportunities in listing ion channel molecules by cell type

Understanding the roles which individual ionic channels play in the complex electrophysiological phenotypes of native† cells remains a significant challenge. The overwhelming range of studies covering aspects of ion channel expression in vertebrate cells offers unique problems when compiling a representative overview. Certainly the linking of specific ion channel gene expression to cell type is a first step towards a more comprehensive indexing, and towards this goal, cell-type-selective studies are useful for a number of reasons. First, they can help visualize the whole range of channel expression by providing an inventory of conductances† observed. Secondly, these studies generally define the experimental conditions required to observe a given conductance. Thirdly, they include much information directly relating specified ionic conductances to the functions of the cell type concerned. Collated information such as this should be of increasing utility in showing the relationship of electrophysiological phenotype to mechanistic information on their gene structure and expression-control (which largely correlates with cell-type lineage). At this time it is difficult to build a definitive catalogue of ion channel gene expression patterns mapped to cell type, not only because the determinants of gene expression are scarcely explored, but also because there remain many unavoidable ambiguities in phenotype definition. Some of these problems are discussed below.

Problems of uneven coverage/omissions

Certain cell preparations have been intensely studied for ion channel expression while others have received very little attention for technical, anatomical or other reasons. Furthermore, a large number of native† ionic currents can be induced or inhibited by agonists† that bind to co-expressed G protein† -coupled receptors†. Thus a difficulty arises in deciding whether channel currents can be unambiguously defined in terms of action at a separately encoded receptor protein. While it is valid to report that an agonist-sensitive current is expressed in a defined cell type, the factors of crosstalk† and receptor-transducer† subtype specificities in signalling systems are complex and may produce an ambiguous classification. Receptor-coupled agonist-sensitivities are an important factor contributing to cell-pharmacological and -electrical phenotype†, but the treatment here has been limited to a number of tabular summaries of ion channel regulation through coupling to G protein-linked effector† molecules (see Resource A – G protein-linked receptors, entry 56). As stated earlier, the entries are not sorted on agonist specificity except where the underlying ion channel protein sequence would be expected to form an integral ionic channel whose gating† mechanism is also part of the assembled protein complex.

Cell preparation methods are variable

A further problem inherent in classifying ion channels by their patterns of expression is that the choice of tissue or cell preparation method may influence phenotype†. The behaviour of channel-mediated ionic currents can be measured in native† cells, e.g. in the tissue slice, which has the advantages of extracellular ionic control, mechanical stability, preserved anatomical location, lack of requirement for anaesthetics and largely undisturbed intercellular communication. Cell-culture techniques show similar advantages, with the important exceptions that normal developmental context, anatomical organization and synaptic arrangements are lost and (possibly as a consequence) the 'expression profile' of receptor and channel types might change. Cultured cell preparations may also be affected by 'de-differentiation†, processes and (by definition) cell lines† are uncoupled from normal processes of cell proliferation†, differentiation† and apoptosis†. Acutely dissociated cells from native† tissue may provide cell-type-specific expression data without anomalies introduced by intercellular (gap junctional) conductances, but the enzymatic or dispersive treatments used may also affect responses in an unknown way.

Verbal descriptions of cell-type expression divisions are arbitrary and are not rigorous

Definitive mapping of specific ion channel subtype expression patterns has many variables. Localization of specific gene products are most informative when in situ localizations are linked to the regulatory factors controlling their expression (see glossary entry on Gene expression-control†). The complexity of this task can extend to processes controlling, for example, developmental regulation, co-expressed protein subunit stoichiometries and subcellular localizations.

Complete integration of all structural, anatomical, co-expression and modulatory data for ion channels could eventually be accommodated within interactive graphical databases which are capable of providing 'overlays' of separately collected in situ expression data linked to functional properties of the molecules. By these methods, new data can be mathematically transformed to superimpose on fixed tissue or cell co-ordinates for comparison with existing database information.

Software development efforts focused on the acquisition, analysis and exchange of complex datasets in neuroscience and mouse development have been described, and the next few years should hopefully see their implementation. For further information, see

Baldock, R., Bard, J., Kaufman, M. and Davidson, D. (1992) A real mouse for your computer, Bioessays 14: 501–2

Bloom, F. (1992) Brain Browser, V 2.0. Academic Press (Software).

Kaufman, M. (1992) The Atlas of Mouse Development, Academic Press

Wertheim, S. and Sidman, R. (1991) Databases for Neuroscience, Nature 354: 88–9

To help rationalize the choices available for selection of these 'prototype' classifications, see Resource H - Listings of cell types, entry 63. These listings may also have some practical use for sorting the subject matter of journal articles into functionally related groups. A proposed integration of information resources relating different aspects of cell-signalling molecule gene expression is illustrated in Fig. 4 of the section headed Feedback CSN access, entry 12.

Field number 09: Channel density

This field should contain information about estimated numbers of channel molecules per unit area of membrane in a specified preparation. This field lists information derived from local patch-clamp 'sampling' or autoradiographic detection in membranes using anti-channel antibodies. The field should also describe unusually high densities of ion channels ('clustering') in specified membranes where these are of functional interest.

Field number 10: Cloning resource

This field should refer to cell preparations relatively 'rich' in channel-specific mRNA (although it should be noted that many ion channel mRNAs are of low abundance†). Otherwise, this field defines a 'positive control' preparation likely to contain messenger† RNA† encoding the channel. Preparations may express only specific subtypes of the channel and therefore related probes (especially PCR† probes) may not work. Alternatively, a genomic† cloning resource may be cited.

Field number 11: Developmental regulation

This field should contain descriptions of ion channel genes demonstrated (or expected to be) subject to developmental gene regulation – e.g. where hormonal, chemical, second messenger† or other environmental stimuli appear to induce (or repress) ion channel mRNA or protein expression in native† tissues (or by other experimental interventions). Protein factors in trans† or DNA structural motifs†in cis† which influence transcriptional activation†, transcriptional enhancement† or transcriptional silencing† should also be listed under this fieldname. Information about the timing of onset for expression should also be included if available, together with evidence for ion channel activity influencing gene activation† or patterning† during vertebrate development.

Field number 12: Isolation probe

This field should include information on probes used to relate distinct gene products by isolation of novel clones following low-stringency cross-hybridization screens†. The development of oligonucleotide† sets which have been used to unambiguously detect subtype-specific sequences by PCR†, RT-PCR† or in situ hybridization† should be identified with source publication. Both types of sequence may be able to serve as unique gene isolation probes, dependent upon the library† size, target abundance†, screening stringency† and other factors.

Field number 13: mRNA distribution

This field should report either quantitative/ semi-quantitative or presence/absence (±) descriptions of specific channel mRNAs in defined tissues or cell types. This type of information is generally derived from Northern hybridization†, RNAase protection† analysis, RT–PCR† or in situ expression assays. See also notes on expression atlases under Field number 08: Cell-type expression index.

Field number 14: Phenotypic expression

This field should include information on the proposed phenotype † or biological roles of specified ion channels where these are discernible from expression studies of native † (wild-type) genes. Phenotypic † consequences of naturally occurring (spontaneous) mutations † in ion channel genes are included where these have been defined, predicted or interpreted (see also Fields 26–32 of the STRUCTURE & FUNCTIONS section for interpretation of site-directed mutagenesis † procedures as well as Resource D - ‘Diagnostic' tests, entry 59). Associations of ion channels with pathological states, or where molecular ‘defects' could be ‘causatory' or contribute to the progression of disease should be listed in this field (for links with established cellular and molecular pathology databases, see Fig. 4 of Feedback & CSN access, entry 12).

The Phenotypic expression field may include references to mutations in other (‘non-channel') genes which affect channel function when the proteins are co-expressed. It is also used to link descriptions of specific (cloned) molecular components to native cell-electrophysiological phenotypes. In due course, this field will be used to hold information on phenotypic & effects of transgenic & manipulations of ion channel genes including those based on gene knockout & or gene locus replacement & protocols.

Field number 15: Protein distribution

This field should report results of expression patterns determined with probes such as antibodies raised to channel primary† sequences or radiolabelled affinity ligands†.

Field number 16: Subcellular locations

This field should describe any notable arrangements or intracellular locations related to the functional role of the channel molecule, e.g. when the channel is inserted into a specified subcellular membrane system or is expressed on one pole of the cell only (e.g. the basolateral† or apical† face).

Field number 17: Transcript size

This field should list the main RNA transcript† sizes estimated (in numbers of ribonucleotides) by Northern† hybridization analysis. Multiple transcript sizes may indicate (i) alternative processing ('splicing†’) of a primary transcript†, (ii) the use of alternative transcriptional start sites†, or (iii) the presence of 'pre-spliced' or 'incompletely spliced' transcripts identified with homologous nucleotide probes† in total cell mRNA† populations. Note that probes can be chosen selectively to identify each of these categories; 'full-length' coding sequence† (exonic†) probes are the most likely to identify all variants, while probes based on intronic† sequences (where appropriate) will identify 'pre-splice' variants.

Criteria for SEQUENCE ANALYSES sections

This section should bring together data and interpretations derived from the nucleic acid or protein sequence of the channel molecule. The symbol [PDTM] denotes an illustrated feature on the channel monomer protein domain topography model, which is presented as a central figure in some entries for sequenced ion channels. These models are only intended to visualize the relative lengths and positions of features on the whole molecule (see the description for field number 30, Predicted protein topography). The PDTMs as presented are highly diagrammatic - the actual protein structure will depend on patterns of folding, compact packing and multi-subunit associations. In particular, the relative positions of motifs, domain shapes and sizes are subject to re-interpretation in the light of better structural data. Links to information resources for protein and nucleic acid sequence data are described in the Database listings field towards the end of each entry.

Field number 18: Chromosomal location

This field should provide a chromosomal locus† designation (chromosome number, arm, position) for channel gene(s) in specified organisms, where this is known. Notes on interactive linking to gene mapping database resources appear under an option of the Cell-Signalling Network 'home page' (see Feedback & CSN access, entry 12).

Field number 19: Encoding

This field should report open reading frame† lengths as numbers of nucleotides or amino acid residues encoding monomeric channel proteins (i.e. spanning the first A of the ATG translational start codon† to the last base of the translational termination codon†). The field should report