12 October 2013 | NewScientist | 45 44 | NewScientist | 12 October 2013

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A fertile, fexible brain couldnt live without its roving band
of caretakers. Moheb Costandi meets the microglia
THE
MIND MINDERS
Y
OUmaynot realiseit, but your
brainis hometoanarmyof invaders.
Ridinginonbloodvessels andnerve
fibres duringyour first months inthewomb,
theseamorphous creatures colonisedevery
part of your brain, wheretheytransformed
intoastrange, tentacledform, beforelying
still, waitinglikespiders at thecentres of
their webs.
It sounds sinister, but these shape-shifters,
knownas microglia, are nocause for alarm.
Rather, theyare anunderappreciatedally.
While neurons, sparkingwithelectrical
activity, steal the limelight as the makers of
thought, we nowknowthat microglia are just
as critical for a fertile, flexible mind.
As master multitaskers, microglia play
manydifferent roles. Onthe one hand, they
are the brains emergencyworkers, swarming
toinjuries andclearingawaythe debris to
allowhealingtobegin. Onthe other hand,
duringtimes of rest, theyare its gardeners
andcaretakers, overseeingthe growthof new
neurons, cultivatingnewconnections and
pruningbackregions that threatento
overgrow. Theymayalsofacilitate learning, by
preparingthe groundfor memories toform.
Inthis way, microglia empower us withthe
abilitytoadapt tolifes experiences. Yet their
workcanbackfire, andneuroscientists now
believe that microglia will provide insights to
neurological conditions suchas Alzheimers
disease andautism.
Our first viewof these mercurial cells
came inthe 1840s, whenpathologist Rudolph
Virchowfirst notedthe presence of microglia
near sites of braininjury. As our
understandingof the brains buildingblocks
grew, theywere later consideredtobe part of
the mysteriousthirdelementof the brain.
The first element was the neuron; the second
was the astrocyte. Astrocytes were thought to
packthe spaces aroundneurons andgive
themnutrients, but nowalsoseemtobe
involvedinother duties, suchas controlling
bloodflowthroughthe brain(see diagram,
page 46).
The thirdelement remaineda puzzle,
however, until the pioneeringworkof Podel
RoHortega inthe 1920s and30s. His stroke of
genius was tofinddyes that allowedhimto
stainthe cells sothat he couldvieweachmore
clearlyunder the microscope. He provedthat
neuroscientists hadpreviouslyconfusedthe
microglia withanother kindof cell, muddying
the picture of their activity. He thenusedhis
microscope totrackthe microglias shape-
shiftinglife cycle includingtheir early
migration, andtheir spidery, dormant state.
AccordingtoRoHortegas observations, it
was onlyinthe face of disease that theystarted
tospringintoactionagain, assumingthe form
of anamoeba tocrawl towards a site of injury.
Once there, theyclearedawaydamage,
devouringmicrobes anddeadneurons to
allowhealingtoset in.
RoHortegas workwouldestablish
microglia as the brains emergencyworkers,
yet it alsocontributedtothe idea that theyare
mostlyinactive under other circumstances,
leadingtheir other duties tobe overlooked
for the rest of the century. If theres a
disturbance, theywill react,says Axel
Nimmerjahn, a biophysicist at the Salk
Institute for Biological Sciences inLa Jolla,
California. The questionis: what dotheydo
inthe healthybrain? Dotheyjust hangout?
Insome ways, the delaywas a matter of
technology. Weve beenaware of microglia
for over 100years, but its beenveryhardto
studythembecause we didnt have the right
tools,says neurobiologist BenBarres of
StanfordUniversityinCalifornia. But inthe
21st centurybetter microscopic techniques
have set the stage for a newunderstanding.
The breakthroughcame 10years ago, while
Nimmerjahnwas doinghis PhDat the Max
PlanckInstitute for Medical Researchin
Heidelberg, Germany. At the time, his team
was more interestedinthose other mysterious
braincells, astrocytes, whichtheyhopedto
markandidentifyusinga chemical dye. But
theywere concernedthat the dye might also
sticktomicroglia, confoundingtheir results.
Sotheyrana follow-upexperiment, injecting
the dye intomice that hadbeengenetically
engineeredsothat their microglia glowed
greenwitha fluorescent protein. Theythen
usedcutting-edge time-lapse imagingto
watchthe mices brains inaction.
The films confirmedthat their dye only
boundtothe astrocytes, as intended. But
althoughtheywere untouchedbythe dye, >
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it was the glowingmicroglia that reallycaught
Nimmerjahns attention. Rather thanlying
dormant, theywere teemingwithactivity,
stretchingout their spider-like tentacles,
calledprocesses, tostroke the nearbyneurons,
synapses andbloodvessels. Theydont just
sit there andwait,he says. Theyactively
patrol their environment.
He showedthe videos tohis colleague,
FrankKirchhoff of the Universityof Saarland
inGermany. I realisedimmediatelythat we
were lookingat somethingspectacular,says
Kirchhoff. Theywere constantlymoving.
The twowrote a paper that is nowconsidereda
landmarkinthe field(Science, vol 308, p1314).
What were the microglia doing? Kirchhoff
andNimmerjahns findings seemedtoshow
microglia inthe midst of some kindof
surveillance. Eachcell sat ina territoryjust
80micrometres across, withits processes
patrollingthe surroundings duringthe course
of 2 to4hours. One possibilitywas that they
were performingregular healthchecks onthe
surroundingneurons todetect disease. But
clues soonbegantoemerge that theywere
lookingfor somethingelse entirely, starting
withthe workof BethStevens, thena postdoc
inBarress labat Stanford.
Once again, it was a surprise finding.
Stevens was examiningthe genetic activityof
the synapse at the time, andshe was puzzled
bya gene that codedfor complement proteins,
a set of molecules normallyassociatedwith
the immune system. These proteins were
knowntocirculate inthe bloodandhelp
antibodies andother immune cells toclear
awaypathogens but what were theydoingat
a synapse inthe healthybrain? As she delved
deeper, she foundthat these complement
proteins were actingas the markof deathfor
the synapse, attractingnearbymicroglia that
wouldthenripout andengulf the neural
connection.
Brain topiary
This findingput microglia at the heart of
the brains synaptic pruningprocess. As our
brains grow, theyformsynaptic connections
somewhat haphazardly. Later on, particularly
duringadolescence, the brainprunes backand
refines the neural circuits sothat theybecome
more efficient at processinginformation. Far
frombeingdormant bystanders, microglia
were shapingandguidingthe brainthrough
one of its biggest upheavals.
Suchtopiarycanalsobringabout the
gentler transformations that helpus adapt
tonewsituations. One of the best pictures of
this process inactioncomes fromMarie-Eve
Tremblay, nowat Laval UniversityinQuebec,
Canada. She reareda groupof mice inthe dark
for a fewdays andthenexposedthemtolight.
All the while, she examinedactivityintheir
visual cortices usinganelectronmicroscope
anda technique calledtwo-photonin
vivoimaging.
As expected, she foundthat manyof the
neurons were rewiringtheir connections in
the changingconditions, particularlyat the
dendritic spine structures that formone side
of the synapse. Whenthe mice were rearedin
the dark, the spines seemedtoshrivel up, but
whenthe lights came on, the spines recovered
andgrewlarger.
Importantly, Tremblayfoundthat microglia
dramaticallyalter their behaviour inresponse
tothese changes. Light deprivationmade
themassociate more closelywithshrinking
spines. Microglial processes not onlytouched
these synapses, but actuallyengulfedthem,
says Tremblay. Incontrast, exposure tolight
hadthe opposite effect the microglia shrank
awayfromthe synapse as it grewand
strengthened(PLoS Biology, vol 8, e1000527).
All inall, it appearedthat the microglia were
closelymonitoringtheir territorytohunt out
andprune weaker connections that were not
beingusedsothat the braincouldrefine its
circuits andreserve resources.
Exactlyhowthe microglia knowwhich
connections are ripe for the chopis a matter
of debate. Stevenss workoncomplement
proteins suggests one possibility: the neurons
themselves maybe usingproteins tomarkthe
synapses that are not pullingtheir weight,
readyfor the microglia tocome inanddothe
dirtywork. But there maybe other
mechanisms, too. For example, active neurons
seemtosignal their healthbyproducinga
substance calledfraktaline, whichmight warn
the microglia toleave its synapses alone.
Whatever the mechanism, it is nowclear
that microglia are busytendingforests of
synapses throughout the brain. I lookat
development, adolescence, youngadults
andagedanimals, andIve seenthe same
Microglia(yellow)
probeneural
connections
(green) ready
for pruning
phenomenoneverywhere,says Tremblay.
Andthe more theylook, the more
neuroscientists are findingthat the
microglias remit goes far beyondpruning
withmanyother activities that might each
contribute tothe brains adaptability. For
instance, besides pruningsynapses, microglia
cultivate their development, bysecreting
nutrients calledgrowthfactors that promote
the sproutingof newneural connections. And
once the synapse is formed, theymaymonitor
andtweakthe receptors that helppass
messages betweentwoneurons. Suchchanges,
dubbedsynaptic plasticity, fine-tune the
communicationacross neural networks, and
are thought tobe a keymechanismfor
learning. Indeed, Tremblayhas foundsigns of
highmicroglial activityinthe hippocampus
a brainregionthat is central tomemory.
Microglia mayalsowatchover the early
stages of braindevelopment tomake sure
it doesnt become overgrownwithneurons.
Earlier this year, researchers at the University
of California, Davis, showedthat microglia
control the size of the cerebral cortexby
regulatingthe numbers of neural stemcells,
whichgive rise toimmature neurons inthe
developingbrain. Examiningthe brains of rats
andmacaque monkeys, the researchers found
that microglia are concentratedinareas at
whichneural progenitor cells divide togive
rise toyoungneurons. Activatingthe
microglia significantlyreducedthe number
of cells, whereas deactivatingor eliminating
themledtoa boom(Journal of Neuroscience,
vol 33, p4216).
This growingrsum of duties is leading
some neuroscientists towonder what happens
whenthese important cells malfunction, and
whether that might helpus tounderstand
certainbraindisorders. The role of microglia
insynaptic pruning, for instance, implicates
theminneurodegenerative diseases such
as Alzheimers. Inthe earliest stages of
Alzheimers, cells begintodie off inthe
hippocampus. This degenerationspreads
throughout the brain, andis accompanied
bythe widespreadloss of synapses.
Alzheimers involves the depositionof
several mutatedproteins withinandaround
braincells, anddeposits of one of these
proteins, calledbeta-amyloid, are widely
believedtobe toxic toneurons. One popular
idea is that microglia might graduallylose
their abilitytomopupthe beta-amyloid
deposits.
But their newlydiscoveredrole insynaptic
pruningsuggests theymaybe involvedin
another way: perhaps the microglia become
over-excitedandstriptoomanysynapses,
says Barres. For instance, the brainmay
produce toomanyof those complement
proteins, inadvertentlytagginglarge swathes
of synapses for destruction. Alongthese lines,
Barres has foundthat complement proteins
doseemtoamass at synapses as we age, while
genetic studies have foundthat a mutationin
a gene involvedinthe taggingprocess canput
people at increasedriskof the disease.
Aclear verdict onanAlzheimers connection
is a longwayoff, however. Alexei Verkhratsky
at the Universityof Manchester, UK, is one
neurophysiologist whowouldlike tosee direct
evidence of hyperactive microglia before he
will be convinced. But if theyare foundtobe
involved, it raises the possibilityof new
therapies. Barres, for instance, has developed
a drugthat targets the complement proteins
andsocouldpotentiallyprevent microglia
fromharmfullyengulfingsynapses.
The role of microglia insynaptic pruning
alsoimplicates theminautism, whichis
associatedwithimproper wiringof neuronal
circuits duringbraindevelopment.
Intriguingly, the microglia of people with
autismdoseemtobehave differently. They
lie closer totheir neighbouringneurons and
theirtentaclesare more likelytobe hugged
aroundthe cells, thoughmuchmore work
will be neededtounderstandwhether this
interactioncontributes tothe symptoms
of autism. Similarly, some promisingleads
suggest that microglia are more active in
people withdepression, potentiallychanging
the chemical signallinginareas of the brain
that are thought toregulate our mood.
It is nowclear that learningmore about
the biologyof microglia couldhelpus gain
a better understandingof bothhealthyand
diseasedbrains. Amongthe most pressing
questions is howmicroglia interact with
neurons andastrocytes. Barres, Stevens and
Tremblayare all workingtoidentifythe
signals microglia use tocommunicate.
The revelationthat restingmicroglia
are anythingbut dormant alsoraises the
possibilitythat these shape-shifters may
exist inmanymore forms thanwe realise.
Activationof microglia is not anall or none
process, but rather a continuumwhich
produces manydifferent phenotypes that
we still dont knowanythingabout,says
Verkhratsky. Andthat couldhave implications
for the whole of neuroscience. Its obvious
that we have toredefine our approachtothe
brain. We needtostopdividingit intoneurons
andglia, because we are dealingwitha tissue
containingmanycell types withdistinct
functions, whichare workingtogether.
Barres agrees that microglia are harbouring
manymore mysteries. Whoknows what
microglia are doingtoneurons interms of
synaptic activity?he says. Indeed, microglia
mayevenplaya direct role inthe electrical
communicationthat makes thought itself.
Were still at the tipof the iceberg. n
MohebCostandi is a sciencewriter inLondon, who
blogs at guardian.co.uk/science/neurophilosophy
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During Alzheimers disease,
microglia may become
over-excited, stripping
away too many synapses
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