Neuropeptides!

OverviewofNeuropeptides:AwakeningtheSenses?

AndrewRusso,PhDMolecularPhysiology&Biophysics

UniversityofIowa

DisclosuresFunding:NIH,DoD,VAMC,AlderBiopharma

Consultant:PharmnovoAlderBiopharmaEliLillyLeClairRyan;Medacorp;Sudler NY;DDBRemedy

Objectives

• Toappreciatethediversityofneuropeptidesinthebody.

• TorecognizethatneuropeptidesactbymodulatingsynaptictransmissionintheCNS.

• Toappreciatethatneuropeptidesalsoactinperipheraltissuestoaltercellularactivities.

http://www.luminousself.com/awakening-the-luminous-self.html

WhatareNeuropeptides?

• Distinctionbetweenneuropeptide andpeptidehormoneissolelybasedonsynthesisinneuron

• Botharesynthesized,modified,anddegradedbysamesetsofenzymesandbothcanactnearbyandatadistance

• Nearlyallneuropeptidesalsofoundaspeptidehormones

• So,whileweusethetermneuropeptide,remember…

• neuropeptidesarenotjustinthenervoussystem–theyactbothinandoutoftheCNS

CandacePert1946-2013http://www.6seconds.org/wp-content/uploads/2007/01/synapse-widened-pert.jpg

“Neuropeptides aresmallprotein-likemolecules(peptides)usedbyneurons tocommunicatewitheachother”(Wikipedia)

DiversityofNeuropeptides

• >100knownneuropeptidesinhumanbrainwww.neuropeptides.nl/tabel%20neuropeptides%20linked.htm

• >1000totalpeptidesinHomosapiens• basedongenomehomologiesofneuropeptides,peptidehormones,cytokines,growthfactors,antimicrobialpeptides,toxins&venompeptides,antifreezeproteins. www.peptides.be/?p=home

• Evolutionaryconservationofpeptidesandreceptors Global view of the evolution and diversity of

metazoan neuropeptide signaling Jekely, 2014 PNAS 110, 8702http://www.pnas.org/content/110/21/8702.full

DiverseNeuropeptideFamiliesConnected tomigraine? Other familiesCGRP:CGRP(a,b),calcitonin,amylin,adrenomedullin (AM1,2) Opioids:enkephalins,dynorphin,endorphins,nociceptin

Somatostatin/cortistatin

Glucagon/secretin:PACAP,VIP,glucagon,secretin,GHRH,GIP Natriureticfactors:ANF,BNF,CNP

GRP,neuromedins

Vasopressin/oxytocin Endothelins

CCK/gastrin

F- andY-amides:NPY,PPY,NPFF Insulins:insulin,IGFs,relaxins

Motilin/ghrelin

Tachykinins:SubP,neurokininA,neuropeptideK,neuropeptidegamma

Galanins

Gonadotropinreleasinghormones

Tensins:angiotensin,neurotensin,bradykinin NeuropeptideB/W/S

Neurexophilins

CRH-related:CRH,urocortin,urotensin Cerebellins

Granins:chromogranins,secretogranins

Adiposeneuropeptides:leptin,adiponectin,resistins

Family-less:orexins, MCH,TRH,PTHrP,CART,AGRP,prolactin,diazepam-bindinginhibitorpeptide,etc http://www.neuropeptides.nl/tabel%20neuropeptides%20linked.htm

CommonFeaturesofNeuropeptides

1. Modulatetargetcells• CNS=enhance/dampensynapticactivity• Peripheraltissues=neurohormones,manytargets,manyfunctions

2. Allaresignaltransducersthatactatcell-surfacereceptors• NearlyallactatGproteincoupledreceptors• Consistentwithneuropeptidesinducingaslowerandmodulatoryresponsecomparedtoneurotransmitters

3. Allprocessedfromprecursors• Synthesistodegradationsharedfeatures

CommonFeatures:Synthesis&Release

DickMains&BettyEipper labhttp://neuropeptidelab.uchc.edu

Processingofprecursors

Fromdensecorevesiclesatsynapses,cellbodies,varicositiesalongaxon.Atsynapsesoftenco-releasedwithaneurotransmitter.

Pro-peptidesareproteolytically cleavedandoftenmodifiedbyC-terminalamidation.

Howmanyneuropeptidesinavesicle?

• Howmanyneuropeptidescandanceontheheadofapin(orinasecretoryvesicle)?

• About100,000peptidesinalargedensecorevesicle

• Calculatedfrom:(3-10mM,Mains+Eipper,1999,BasicNeurochemistry,6th ed;60mM foroxytocin,vasopressininposteriorpit,Nordmann +Morris,1984,PNAS81:180-84)

• 100-200nmdiameter,vol =2x106 cubicnm,2x10-18 liter/vesicle• 10mmol/lx2x10-18 l=20x10-18 mmol =2x10-20 mol/vesicle• 2x10-20 mol x6x1023 molec/mol=1.2x104 molec/vesicle(~100,000peptides/vesicle)

• Tenstohundredsofvesiclesreleasedatsynapse,cellbodies,varicositiesintimescaleofmsec (chromaffincellshave20,000densecorevesicles,Plattner,JCB,1997)

• Somillionsofneuropeptidesarereleasedinashortburstfromasinglecell

CommonFeatures:Diffusion&Degradation

• Neuropeptidesdiffusefrompointofrelease(volumetransmission)

• Nonsynaptic dispersioninextracellularfluidandCSF• Slowlyremovedfromextracellularspace,whichcontributestorelativelylonglastingeffects

• Noreuptakesystem• Canbeactiveatrelativelylargedistancesatrelativelylowconcentrations

• Highreceptoraffinities(comparedtoneurotransmitters)

• Inactivationoccursbyextracellularproteases• Thiscanalsocreatedifferentbioactivepeptides

VandenPol(2012)Neuron76:98–115

Neuropeptides&SensoryPerception?

• Neuromodulationinthebraincoupledwithalteredperipheralsignals

http://www.epainassist.com/brain/sensory-processing-disorder-spd

http://www.luminousself.com/awakening-the-luminous-self.html

http://neuroscience.uth.tmc.edu/s1/chapter14.html

WithintheBrain:Neuromodulation• Generallymodulateactivityofco-releasedneurotransmitterstoeitherincreaseor

decreasestrengthofsynapticsignaling• Influencemanyfunctions,e.g.analgesia,reward,foodintake,metabolism,reproduction,

socialbehaviors,learningandmemory.

WithinthePeriphery:AlterCellularActivities

• Canfunctionsimilartopeptidehormonesandactonvarioustargettissuestomodulatenearlyallbodilyfunctions

• Atthevasculature

Edvinsson and Uddman, Brain Research Reviews 48(3):438-56 2005

NeuropeptideFamilies&MigraineConnected tomigraine? Other families

CGRP:CGRP (a,b),calcitonin,amylin,adrenomedullin (AM1,2) Opioids:enkephalins,dynorphin,endorphins,nociceptin

Somatostatin/cortistatin

Glucagon/secretin:PACAP,VIP,glucagon,secretin,GHRH,GIP

Natriureticfactors:ANF,BNF,CNP

GRP,neuromedins

Vasopressin/oxytocin Endothelins

CCK/gastrin

F- andY-amides:NPY,PPY,NPFF Insulins:insulin,IGFs,relaxins

Motilin/ghrelin

Tachykinins:SubP,neurokininA,neuropeptideK,neuropeptidegamma

Galanins

Gonadotropinreleasinghormones

Tensins:angiotensin,neurotensin,bradykinin NeuropeptideB/W/S

Neurexophilins

CRH-related:CRH,urocortins,urotensins Cerebellins

Granins:chromogranins,secretogranins

Adiposeneuropeptides:leptin,adiponectin,resistins

Family-less:orexins, MCH,TRH,PTHrP,,CART,AGRP,prolactin,diazepam-bindinginhibitorpeptide,etc http://www.neuropeptides.nl/tabel%20neuropeptides%20linked.htm

NPY:PremonitoryPhaseFeedingPeptides&Migraine

• NPY Roles- Pain- Appetite control- Synchronization of circadian rhythm

• NPY brain location- Hypothalamic nuclei- Basal ganglia- Limbic system

• NPY receptors (G protein coupled Y receptors)- Six sub-types- Not all in humans (1-5)

• In migraine- Could neuropeptide Y mechanisms link altered appetite and nociceptive processing?

SlidemodifiedfromPeterGoadsby

NPY &Trigeminovascular System

Martins-Oliveira et al.,Pain 2016;167:1666

• Anesthetized rat; durovascular stimulation• Recording from second order trigeminocervical neurons• NPY dose-dependently inhibits nociceptive trigeminovascular transmission

SlidefromPeterGoadsby

CRH&Migraine?

Robertetal.,2013,JNeurosci 33:8827Noseda andBurstein,2013,Pain154:S44 Lietal.,2016,Cell167:60

• CRHisastresshormonereleasedbythePVH,whichsendsprojectionstospinaltrigeminalnucleus

• Sexualdimorphism.CRHlevelsinPVHaretwiceashighinfemale&actionsareinhibitedbyoxytocininmales,butnotfemales

CRHlinkswithCGRPandPACAP

• CGRPinjectioninPVHinducesCRH-dependentreleaseofACTH,corticosterone• PACAPinjectioninPVHactivatesspinaltrigeminalnucleus

Robertetal.,2013,JNeurosci 33:8827

CGRPinjection

Dhillo etal.,2003,Endocrinology144:1420

PACAPinjection

Conclusion

• GiventheevidenceforCGRPandtheemergingevidenceforotherpeptides…

• Itseemslikelythatalteredneuropeptideactionsmaybeageneralthemeunderlyingtheheightenedsensorystateofmigraine http://io9.gizmodo.com/5752351/zapp

ing-the-brain-sparks-bright-ideas

Adisa KuburasBrandon ReaBianca MasonLevi SowersAnne-Sophie WattiezConnor CastonguayBennett Robertson

Collaborators:• Randy Kardon (U Iowa)• Pieter Poolman (U Iowa)• Leon Garcia-Martinez (Alder)• Minyan Wang (Xi’an Jiaton-

Liverpool Univ)• Mark Chapleau (U Iowa)

Former membersPaul DurhamAna RecoberEric Kaiser

Acknowledgments