Serotonin Receptor Technologies

247 Pages·2015·7.905 MB·English

by  Wolfgang Blenau, Arnd Baumann (eds.)

Neuromethods 95
Wolfgang Blenau
Arnd Baumann Editors
Serotonin
Receptor
Technologies
N
EUROMETHODS
Series Editor
Wolfgang Walz
University of Saskatchewan
Saskatoon, SK, Canada
For further volumes:
h ttp://www.springer.com/series/7657

Serotonin Receptor Technologies
Edited by
Wolfgang Blenau
Universität zu Köln, Köln, Germany
Arnd Baumann
Forschungszentrum Jülich, Jülich, Germany
Editors
Wolfgang Blenau Arnd Baumann
Universität zu Köln Forschungszentrum Jülich
Köln, Germany Jülich , Germany
ISSN 0893-2336 ISSN 1940-6045 (electronic)
ISBN 978-1-4939-2186-7 ISBN 978-1-4939-2187-4 (eBook)
DOI 10.1007/978-1-4939-2187-4
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Series P reface
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v

Prefa ce
Serotonin (5-hydroxytryptamine, 5-HT) is an important signaling molecule, which plays
a key role in regulating and modulating physiological and behavioral processes in both
deuterostomes (e.g., mammals) and protostomes (e.g., fl atworms, nematodes, and
arthropods). In the central nervous system (CNS), serotonin modulates mood, percep-
tion, reward, anger, aggression, appetite, memory, sexual behavior, and attention. In
addition, serotonin has important functions outside the CNS, including the regulation of
energy balance and food intake, gastrointestinal and endocrine function, and cardiovas-
cular and pulmonary physiology. Thus it is not surprising that impairment of the seroto-
nergic system has been implicated in the pathogenesis of several human diseases like
depression, schizophrenia, anxiety and panic disorders, migraine, hypertension, eating
disorders, vomiting, and irritable bowel syndromes.
Similar to other biogenic amines, serotonin is synthesized from an amino acid precur-
sor. Two enzymatic steps, catalyzed by the enzymes tryptophan hydroxylase and
3,4-d ihydroxyphenylalanine (DOPA) decarboxylase, are necessary to transform tryptophan
to the primary amine serotonin. Specifi c functions of serotonin result from its binding to
and subsequent activation of membrane receptors. Uncovering the precise molecular
mechanism of serotonin signaling is complicated by the fact that humans express 14
serotonin- receptor subtypes. One of these belongs to the family of cys-loop ligand-gated
ion channels (5-HT ) whereas all other proteins are G-protein-coupled receptors (GPCRs).
3
In mammals, serotonin-binding GPCRs fall into six distinct classes, with some classes con-
taining several receptor subtypes. Additional 5-HT receptor heterogeneity originates from
alternative splicing of primary transcripts, RNA editing, and potential formation of receptor
heterodimers. Activation of 5-HT receptors causes transient changes in the concentration
of intracellular messengers (e.g., cAMP, Ca 2+ ), ion channel activity, or reaction cascades that
may lead to changes in gene regulation. The reuptake of serotonin from the synaptic cleft
into the presynaptic terminus by the Na + -dependent serotonin transporter (SERT) termi-
nates the action of serotonin and allows functional recycling of the neurotransmitter.
Notably, SERT is the target of antidepressant medications, e.g., sertraline, paroxetine, cita-
lopram, and escitalopram, which serve in keeping the extracellular concentration of sero-
tonin elevated.
In recent years, a vast amount of new information has been accumulated concerning
the functional characteristics of the various 5-HT receptor subtypes and the SERT. Data are
based on two main lines of research: (1) operational pharmacology applying selective
ligands and (2) molecular biological tools. It is also worth mentioning that serotonin works
in concert with other neurotransmitter systems, and crosstalk of signaling mechanisms is
key to understanding various basic physiological functions and pathological states. To pro-
vide readers with state-of-the-art methodologies currently applied in serotonin research, we
have assembled a collection of protocols provided by leading experts in the fi eld. The
approaches described in this volume vary from molecular biological and biochemical tech-
niques (e.g., regarding receptor dimerization), fl uorescence microscopy and imaging
applications, fl ow cytometry, the use of organotypic slice and cell cultures to the generation
vii
viii Preface
of genetically modifi ed animal models and the development of sophisticated behavioral
tests, thus covering a wide spectrum of techniques to study serotonergic signaling in detail.
Compared to mammals, the serotonergic system of protostomian phyla (e.g., fl at-
worms, nematodes, and arthropods) is slightly less complex. However, similarities in modes
of drug action, behavioral responses, and gene activity patterns between protostomian and
mammalian serotonergic systems open alternative routes to studying and understanding
fundamental neuropharmacological processes that are relevant to human diseases. In this
context, simpler model organisms for which various analytical tools have been established
allow high-throughput analyses at signifi cantly reduced overall costs and thus provide
promising perspectives for future serotonin research. For example, C aenorhabditis elegans
is excellently suited to analyze serotonergic signaling because locomotion is regulated by
serotonin in this nematode. Thus, some invertebrate models currently used in serotonin
research are also covered in this volume.
Many of the experimental procedures described in this NEUROMETHODS volume
will be also valuable for researchers working on similar problems with related GPCRs and
neurotransmitter transporters. Therefore, we are confi dent that this collection of methods
will foster both basic and translational research aiming to further deepen our understanding
of the various facets aminergic systems provide. Last but not least, we would like to take this
opportunity to express our sincere gratitude to all the authors for their efforts providing
their valuable contributions to this volume of the NEUROMETHODS series.
Köln, Germany Wolfgang Blenau
Jülich, Germany Arnd Baumann
Contents
Series Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi
PART I RECEPTOR DIMERIZATION
1 Novel Approaches to Serotonin Receptor Interaction Studies . . . . . . . . . . . . . 3
Sylwia Łukasiewicz , Ewa B łasiak , Kinga S zafran-Pilch ,
and Marta Dziedzicka-Wasylewska
2 T echniques for the Study of GPCR Heteromerization in Living Cells
and Animal Models. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
José L . M oreno , J eremy Seto , J ames B . H anks , and Javier G onzález-Maeso
PART II M ANIPULATION AND ANALYSIS OF SEROTONERGIC
SIGNALING IN BRAIN SLICES
3 Organotypic Slices and Biolistic Transfection for the Study
of Serotonin Receptor Function in CNS Neurons . . . . . . . . . . . . . . . . . . . . . . 3 9
Kelly M cGregor , J ean-Claude Beïque , and Rodrigo Andrade
PART III MOUSE MODELS TO STUDY SEROTONIN-ASSOCIATED
BEHAVIORAL DISORDERS
4 Dissecting a Model of Depressive-Related Phenotype
and Antidepressant Effects in 129S2/SvPas Mice . . . . . . . . . . . . . . . . . . . . . . 5 9
Silvina L. Diaz and Luc M aroteaux
5 T he Murine Serotonin Syndrome and the 5-HT Receptor:
1A
Behavioral Effects and Hypothermia. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
Robert H aberzettl , Heidrun F ink , Silke D ietze , and Bettina B ert
6 5 -HT Receptor Subtype, β-Arrestin Level, and Rapid-Onset
4
Effects of Antidepressant Drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 01
Indira M endez-David , Denis Joseph David , J ean-Philippe Guilloux ,
René Hen , and A lain M ichel G ardier
PART IV SEROTONIN TRANSPORTER
7 Autoshaping Memory Formation and Retention Loss:
Are Serotonin and Other Neurotransmitter Transporters Involved?. . . . . . . . . 1 25
Alfredo Meneses and R uth T ellez
8 Flow Cytometry to Determine Serotonin Transporter Function
in Human Peripheral Blood Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 51
Brendan S . Beikmann and Anne M . A ndrews
ix