Brain-Derived Neurotrophic Factor (BDNF)

254 Pages·2019·6.981 MB·English

by  Carlos B. Duarte, Enrico Tongiorgi

Neuromethods 143
Carlos B. Duarte
Enrico Tongiorgi
Editors
Brain-Derived
Neurotrophic
Factor
(BDNF)
N
EUROMETHODS
SeriesEditor
Wolfgang Walz
University ofSaskatchewan
Saskatoon, Canada
Forfurther volumes:
http://www.springer.com/series/7657
Brain-Derived Neurotrophic
Factor (BDNF)
Edited by
Carlos B. Duarte
Center for Neuroscience and Cell Biology and Department of Life Sciences, University of Coimbra,
Coimbra, Portugal
Enrico Tongiorgi
BRAIN Center for Neuroscience, Department of Life Sciences, University of Trieste, Trieste, Italy
Editors
CarlosB.Duarte EnricoTongiorgi
CenterforNeuroscienceandCellBiology BRAINCenterforNeuroscience
andDepartmentofLifeSciences DepartmentofLifeSciences
UniversityofCoimbra UniversityofTrieste
Coimbra,Portugal Trieste,Italy
ISSN0893-2336 ISSN1940-6045 (electronic)
Neuromethods
ISBN978-1-4939-8969-0 ISBN978-1-4939-8970-6 (eBook)
https://doi.org/10.1007/978-1-4939-8970-6
LibraryofCongressControlNumber:2019934806
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Series Preface
Experimental life sciences have two basic foundations: concepts and tools. The Neuro-
methodsseriesfocusesonthetoolsandtechniquesuniquetotheinvestigationofthenervous
system and excitable cells. It will not, however, shortchange the concept side of things as
carehasbeentakentointegratethesetoolswithinthecontextoftheconceptsandquestions
underinvestigation.Inthisway,theseriesisuniqueinthatitnotonlycollectsprotocolsbut
also includes theoretical background information and critiques which led to the methods
andtheirdevelopment.Thusitgivesthereaderabetter understandingoftheoriginofthe
techniquesandtheirpotentialfuturedevelopment.TheNeuromethodspublishingprogram
strikes a balance between recent and exciting developments like those concerning new
animal models of disease, imaging, in vivo methods, and more established techniques,
including, for example, immunocytochemistry and electrophysiological technologies. New
traineesinneurosciencesstillneedasoundfootingintheseoldermethodsinordertoapply
acriticalapproachtotheir results.
Under the guidance of its founders, Alan Boulton and Glen Baker, the Neuromethods
serieshasbeenasuccesssinceitsfirstvolumepublishedthroughHumanaPressin1985.The
seriescontinuestoflourishthroughmanychangesovertheyears.Itisnowpublishedunder
theumbrellaofSpringerProtocols.Whilemethodsinvolvingbrainresearchhavechangeda
lot since theseriesstarted, thepublishingenvironmentand technologyhavechanged even
more radically. Neuromethods has the distinct layout and style of the Springer Protocols
program,designedspecificallyfor readabilityandeaseofreferenceinalaboratorysetting.
Thecarefulapplicationofmethodsispotentiallythemostimportantstepintheprocess
of scientific inquiry. In the past, new methodologies led the way in developing new dis-
ciplines in the biological and medical sciences. For example, Physiology emerged out of
Anatomyinthenineteenthcenturybyharnessingnewmethodsbasedonthenewlydiscov-
eredphenomenonofelectricity.Nowadays,therelationshipsbetweendisciplinesandmeth-
ods are more complex. Methods are now widely shared between disciplines and research
areas. New developments in electronic publishing make it possible for scientists that
encounter new methods to quickly find sources of information electronically. The design
of individual volumes and chapters in this series takes this new access technology into
account. Springer Protocols makes it possible to download single protocols separately. In
addition, Springer makes its print-on-demand technology available globally. A print copy
canthereforebeacquiredquicklyandforacompetitivepriceanywhereintheworld.
WolfgangWalz
v
Preface
ThepioneeringworkofRitaLevi-Montalcini,VictorHamburger,andStanleyCohenledto
the discovery of nerve growth factor (NGF) as the molecule involved responsible for the
trophic effects of innervated tissues during development of sympathetic and sensory neu-
rons. Laterstudies allowed theidentification ofother membersofthe neurotrophinfamily
oftrophicfactors:brain-derivedneurotrophicfactor(BDNF),neurotrophin-3(NT-3),and
neurotrophin-4/5 (NT-4/5). Neurotrophins have been shown to play important roles in
the control of proliferation, differentiation, survival, and death of neuronal and
non-neuronalcells,anddysregulationofthesemechanismshasbeenassociatedwithdiffer-
entdisorders.
TheeffectsofBDNFhavebeendescribedinthecentralnervoussystemaswellasinthe
peripheralnervoussystem.TheexpressionofBDNFisregulatedbyneuronalactivity,anda
precursor formoftheprotein(proBDNF)isfirstsynthesizedintheendoplasmicreticulum
and later transported to the Golgi apparatus. The intracellular trafficking of the precursor
and mature forms of BDNF has been studied to a large extent in hippocampal neurons,
wheretheneurotrophinisreleasedbyaCa2+-dependentmechanism,fromthepostsynaptic
region and from the axon terminal. BDNF acts through activation of presynaptic and
postsynaptic TrkB receptors, and the complex BDNF-TrkB activates different intracellular
mechanisms (Ras/Erk, Phosphoinositide 3-kinase [PI3-K]/Akt, and phospholipase C-γ
pathways), which have local regulatory roles. However, the BDNF-TrkB complexes may
alsotravelwithinthecellandregulateneuronalactivityinsubcellularcompartmentslocated
far away from the region where the neurotrophin was released. All these steps, from the
regulationofgeneexpressiontotheregulationofBDNFrelease,andthedifferentresponses
tostimulationofTrkBreceptors,arethesubjectofcurrentinvestigation.
BDNF is a key synaptic regulator, both during the synaptogenesis period and after
differentiation, acting on excitatory and inhibitory contacts. The effects of BDNF on
excitatorysynapseshavebeenlargelyinvestigatedinthehippocampus,whereitstrengthens
neuronal communication under specific conditions of activity. These forms of synaptic
plasticity in the hippocampus and in other brain regions are thought to underlie learning
andmemoryformation.Accordingly,BDNFhasbeenshowntoplayaroleincertainforms
oflearningandmemory.
BDNF also plays a role in various disorders of the nervous system, such as depression,
schizophrenia, obsessive-compulsive disorder, Alzheimer’s disease, Huntington’s disease,
Rett syndrome, Down syndrome, epilepsy, and dementia, as well as anorexia nervosa and
bulimia nervosa; neuroprotective effects of BDNF in brain ischemia were also reported.
Furthermore, BDNF-TrkB signaling was shown to contribute to oncogenesis in different
typesoftumors,butanupregulationoftheneurotrophinlevelsinthehypothalamussetsin
motion an anti-tumor immune response. Numerous ongoing studies aim at determining
how the deregulation of the expression, synthesis, intracellular trafficking, and release of
BDNF,aswellastheresponsesmediatedbyTrkBreceptors,areassociatedwithdiseasesof
the nervous system. The use of peripheral BDNF as a biomarker of disease has also been
proposed.
vii
viii Preface
Besides the large number of existing studies on BDNF, the literature accumulating
over the last decades has often produced conflicting results. The ensuing debate indicated
that discrepancies between studies are in part accounted by differences in the methods
used in different laboratories. The peculiar molecular characteristics, the complex cellular
regulation, and the multitude of physiological effects of BDNF make its accurate analysis
prone to artifacts if not carried out with precise methodologies, a prerequisite to obtain
robustdata.
ThepresentvolumeofNeuromethodsdedicatedtoBDNFaimsatprovidinganoverview
of the methodologies currently used in the field to study the physiology of this neurotro-
phin, from the regulation of gene expression to its release and the signaling by TrkB
receptors, as well as in the characterization of the neuronal responses to BDNF and the
role of the neurotrophin in different diseases. We trust this book will help researchers to
furtherexplorethediversityofphysiologicalrolesofBDNF.Finally,wewouldliketothank
allcontributorsforsharingthedetailedprotocolsusedintheirlaboratories.
Coimbra,Portugal CarlosB.Duarte
Trieste,Italy EnricoTongiorgi
Contents
SeriesPreface ................................................................ v
Preface ..................................................................... vii
Contributors................................................................. xi
PART I INTRODUCTION
Brain-DerivedNeurotrophicFactorandtheAttivit`aplasticadeineuroni:
TheNeuronalPlasticityasDefinedbyErnestoLugaro(1870–1940).............. 3
HeatherBowlingandMosesV.Chao
PART II TRANSCRIPTS OF BDNF
UsageofBacterialArtificialChromosomesforStudyingBDNFGene
RegulationinPrimaryCulturesofCorticalNeuronsandAstrocytes............... 13
KaurJaanson,AngelaPa€rn,andT˜onisTimmusk
DetectingSingleandMultipleBDNFTranscriptsbyInSitu
HybridizationinNeuronalCulturesandBrainSections.......................... 27
AndreaColliva,KristenR.Maynard,KeriMartinowich,
andEnricoTongiorgi
StudyingBDNF/TrkBSignaling:TranscriptomeAnalysis
fromaLimitedNumberofPurifiedAdultorAgedMurineBrainNeurons ......... 55
ChinnavuthVatanashevanopakorn,AmitGrover,ArupR.Nath,KevinClark,
PaulSopp,ClausNerlov,andLilianaMinichiello
StudyingBDNF/TrkBSignaling:High-ThroughputMicrofluidic
GeneExpressionAnalysisfromRareorLimitedSamplesofAdult
andAgedCentralNeurons ................................................... 77
ArupR.Nath,RoyDrissen,FeiGuo,ClausNerlov,
andLilianaMinichiello
PART III PROTEIN FORMS OF BDNF
DetectingBDNFProteinFormsbyELISA,WesternBlot,
andImmunofluorescence..................................................... 89
StefanoDoneg`aandEnricoTongiorgi
MethodologyforDetectingandTrackingBrain-Derived
NeurotrophicFactorComplexesinNeuronsUsing
SingleQuantumDots........................................................ 105
AnkeVermehren-Schmaedick,ThomasJacob,andTaniaQ.Vu
ix
x Contents
RecordingActivity-DependentReleaseofBDNF
fromHippocampalNeurons.................................................. 119
TanjaBrigadski,PetraLichtenecker,andVolkmarLessmann
PART IV RECEPTORS OF BDNF
UltrastructuralLocalizationofBDNFandtrkBReceptors........................ 133
ChiaraSalioandAdalbertoMerighi
AnalysisofTrkBReceptorActivityUsingFRETSensors ......................... 149
CharlesE.Hall,JamesO.McNamara,andRyoheiYasuda
PART V SIGNALING CASCADES OF BDNF
BDNF-InducedIntracellularSignaling......................................... 161
Joa˜oR.Gomes,AndreaLobo,CarlosB.Duarte,andMa´rioGra˜os
AMicrofluidicCulturePlatform forNeurotrophinSignalingStudies .............. 185
RuiO.Costa,TaˆniaPerestrelo,DiogoTome´,
andRamiroD.Almeida
PART VI BDNF-INDUCED PROTEIN SYNTHESIS
AND SYNAPTIC REGULATION
BDNF-InducedLocalProteinSynthesisinSynaptoneurosomes
AssessedwithClick-iTL-Azidohomoalanine.................................... 205
VictorBrizandMichelBaudry
ProteomicToolstoStudytheEffectofBDNFonDeNovo
ProteinSynthesis............................................................ 217
HeatherBowlingandEricKlann
BDNFFunctioninLong-TermSynapticPlasticity
intheDentateGyrusInVivo:MethodsforLocalDrugDelivery
andBiochemicalAnalysisofTranslation........................................ 241
DebabrataPanjaandCliveR.Bramham
Index ...................................................................... 257
Contributors
RAMIROD.ALMEIDA (cid:1) CNC-Center forNeuroscienceandCellBiology,Universityof
Coimbra,Coimbra,Portugal;InstituteforInterdisciplinaryResearch,Universityof
Coimbra,Coimbra,Portugal;DepartmentofMedicalSciences,InstituteofBiomedicine-
iBiMED,UniversityofAveiro,Aveiro,Portugal
MICHELBAUDRY (cid:1) GraduateCollegeofBiomedicalSciences,WesternUniversityofHealth
Sciences,Pomona,CA,USA
HEATHERBOWLING (cid:1) CenterforNeuralScience,NewYorkUniversity,NewYork,NY,USA;
KlannLaboratory,Center forNeuralScience,NewYorkUniversity,NewYork,NY,USA
CLIVE R.BRAMHAM (cid:1) DepartmentofBiomedicineandKGJebsenCenter forResearchon
NeuropsychiatricDisorders,UniversityofBergen,Bergen,Norway
TANJABRIGADSKI (cid:1) DepartmentofInformaticsandMicrosystemsTechnology,Universityof
AppliedScienceKaiserslautern,Kaiserslautern, Germany
VICTOR BRIZ (cid:1) DepartmentofMolecularNeuropathology,CentrodeBiologı´aMolecular
SeveroOchoa,CSIC-UniversidadAut(cid:2)onomadeMadrid,Madrid,Spain
MOSESV.CHAO (cid:1) DepartmentofCellBiology,Physiology,andNeuroscienceandPsychiatry,
SkirballInstituteofBiomolecularMedicine,NewYorkUniversityLangoneMedicalCenter,
NewYork,NY,USA
KEVIN CLARK (cid:1) MRCMolecularHaematologyUnit,WeatherallInstituteofMolecular
Medicine,UniversityofOxford,Oxford,UK
ANDREACOLLIVA (cid:1) BRAINCenterforNeuroscience,DepartmentofLifeSciences,University
ofTrieste,Trieste,Italy
RUIO.COSTA (cid:1) CNC-Center forNeuroscienceandCellBiology,UniversityofCoimbra,
Coimbra,Portugal;InstituteforInterdisciplinaryResearch,UniversityofCoimbra,
Coimbra,Portugal
STEFANODONEGA` (cid:1) BRAINCenterforNeuroscience,DepartmentofLifeSciences,University
ofTrieste,Trieste,Italy
ROYDRISSEN (cid:1) MRCMolecularHaematologyUnit,WeatherallInstituteofMolecular
Medicine,UniversityofOxford,Oxford,UK
CARLOSB.DUARTE (cid:1) CNC-CenterforNeuroscienceandCellBiologyandDepartmentofLife
Sciences,UniversityofCoimbra,Coimbra,Portugal
JOA˜OR.GOMES (cid:1) InstitutodeInvestigac¸a˜oeInovac¸a˜oemSau´de(I3S),UniversityofPorto,
Porto,Portugal;MolecularNeurobiology,IBMC-InstituteforMolecularandCellBiology,
UniversityofPorto,Porto,Portugal
MA´RIO GRA˜OS (cid:1) CNC-Center forNeuroscienceandCellBiology,UniversityofCoimbra,
Coimbra,Portugal
AMITGROVER (cid:1) MRCMolecularHaematologyUnit,WeatherallInstituteofMolecular
Medicine,UniversityofOxford,Oxford,UK
FEIGUO (cid:1) DepartmentofPharmacology,UniversityofOxford,Oxford,UK
CHARLESE.HALL (cid:1) DepartmentofPharmacology,DukeUniversitySchoolofMedicine,
Durham,NC,USA
KAURJAANSON (cid:1) DepartmentofChemistryandBiotechnology,TallinnUniversityof
Technology,Tallinn,Estonia
xi