Evolutionofvisionanditsrelationshiptooculardevelopment(EvoDevo)

Morethananyotherorgan,theeyehasshapedtheevolutionofanimalsandecosystems
datingbacktotheCambrianexplosion.Inanimals,eyesusepinholes,lenses,mirrorsand
scanningdevicesinvariouscombinations(seeLandandNilsson,2002).Notalleyesare
pairedandplacedonthehead.Evenacomplexnervoussystemisnotessentialfortheir
developmentastheboxjellyfishhascameratypeeyesconnectedtoasimpleringshaped
nervoussystem.Eyescanbefoundinunicellularalgaes(somedinoflagellatas)withalens
andaretinalikestructureallwithinonecell(seeGehring,2004).

Thefunctionoftheeyeadherestospecificlawsofphysicsuniquetovision.Thehuman
cameratypeeyecontainsthreeopticallytransparenttissues,thecornea,lensandretina.Light
refractiondependsongradualchangesintherefractiveindexoftheocularlens.Theamount
oflightthatcanreachtheretinaisregulatedbythecontraction/expansionoftheiris.
Belongingtoafamilyofsensoryneurons,retinalphotoreceptorsconvertlightenergyinto
electricalsignalstransmittedbytheopticnervetothebrain.Ocularopticsdependsonthe
precisedistancebetweenthelensandretinathatispartiallyregulatedbyintraoculareye
pressure.Specializedtrabecularmeshworkcellsintheanteriorsegmentoftheeyegenerate
intraocularpressure.

Theevolutionofthevertebrateeyehaslongservedasoneofthemostintriguingproblemsof
modernevolutionaryanddevelopmentalbiology.WhenDarwinthoughtofhistheoryof
evolution,herecognizedpotentialdifficultiesinitsapplicationtotheproblemofeye
evolution.InTheOriginofSpecies,Darwinsummarizedhisconcernsinthechapter
DifficultiesoftheTheoryinwhichhediscussesOrgansofExtremePerfectionand
Complication.Modernbiologyhasmadeenormousprogressinitsefforttoexplainthe
processofeyeevolution.Datafromgenetic,biochemicalandembryologicalstudieshave
providedthecurrentgenerationofresearchersanindelibleamountofinformationtoanalyze.
Here,wewillbringourfocustorecentgeneticsstudiesthatarebeginningtogreatlyenrich
ourunderstandingoftheevolutionoftheeye.

Thesubjectofeyeevolutiongainedsignificantmomentumduetothepioneeringstudiesof
W.GehringandhiscoworkersattheUniversityofBasel(seeGehringandIkeo,1999;
Gehring,2004,2005)Anumberofadditionalexcellentreviewarticleshaverecentlycovered
thistopic(Arnheiter,1998;Baileyetal.2004;Fernald,2000,2004;Kozmik,2005;Landand
Nilsson,2002;Nilsson,2004;PiatigorskyandKozmik,2004;TomarevandPiatigorsky,1997;
TrueandCaroll,2002;Wistow1993,1995).Similarly,anumberofmonographshave
examinedgeneticnetworksrelevanttoeyedevelopment(seeDavidson,2000;Wilkins2001;
SchlosserandWagner2004).


StudiesofthesimplestvisualorgansareconsistentwiththeDarwinianprototypeeye.Very
primitivevisionsystems,suchasthoseofthelarvaltrematodeworm,consistonlyofasingle
photoreceptorcellandasurroundingpigmentcell(seeGehringandIkedo,1999).Theworm
senseslightasaresultofthecoordinatedactionsofbothcells:thepigmentedcellscreensthe
photoceptercellsaccesstolightbyvaryingdegreestoenablethelattersabilitytodetect
fromwherethelightiscoming(seeArnheiter,1998;NilssonandLand,2002).Slightlymore
complexvisionsystemsbelongtocertainformsofplanariansthatexhibitthree
photoreceptorsalongsideonepigmentedcell(seeGehring,2004).Theexistenceofsuch
distinctvisualsystemsrangingfromsystemsincapableofprovidingspatialvisiontothose
capableoflightdetection,andfinallytoeyeswithcomplexcentralnervoussystemsprovides
ampleopportunitytoinvestigatethereasonsfortheirvaryinglevelsofcomplexity.An
analysisofthisinformationshouldenrichexistingtheoriesonevolutionandbroadenour
understandingofoculardevelopmentinanumberofmodelsystems.

Herein,wewillfocusonthreeaspectsofeyeevolutionrelatedtoourongoingworkto
understandlensdevelopment:

1. Whataretheessentialproteinsforvisionandfromwheredotheyoriginate?
2. Evolutionfromtheprototypeancestoreyetothepresentdaycomplexeyefoundin
vertebrates.
3. Whatarethegenesthatregulatetheformationoftheeyeandhowdothesegenes
functionindifferentspecies?

1.Whataretheessentialproteinsforvisionandfromwheredotheyoriginate?

Herein,wesummarizestudiesthatshowthatmoleculesessentialforvisualsystemsin
variousanimallineagesarebothstructurallyandfunctionallyrelatedtoproteinsfoundin
othertissuesandorgans.
Thekeymoleculeforvisionisrhodopsinbecauseitisthemoleculethatdetectslight.
Rhodopsinisaseventransmembrane(7TM)proteinhighlyexpressedinretinal
photoreceptors.Microbialopsins(bacteriorhodopsins),withstructuralbutnotsequence
homologywithvisualrhodopsins,arefoundincertainprotistswheretheyfunctionaslight
drivenprotonpumps(seeGehring,2004;2005).Thereissignificantstructuralhomology
betweenbacteriorhodopsinsandchemoattractantreceptorsregulatedbycAMP.Thereare
additionalphotosensitivemoleculesinunicellularalgaeincludingcryptochromes,
phototropsinsandphotoactivatedadenylylcyclases(seeNilsson,2004).Inadditionto
photoreceptors,otherphotosensitivecells,suchasretinalganglioncellsthatsetthecircadian
clock,arepresentintheretina.Inphotosensitiveiriscellsandhypothalamicneurons,
melanopsinsdetectlight(seeArnheiter,1998).Genesencodingrhodopsinsandothergenes

encoding7TMproteinscanbeanalyzedfortheirevolutionaryoriginanddistributionin
individualspeciesthatdoordonotpossesseyes.

Twosignaltransductioncascadesoperatedownstreamofrhodopsin.Arhabdomerictypeof
photoreceptorusesphospholipaseCinthecascade.Inthisinstance,lightcauses
depolarizationofthephotoreceptormembranepotential.Thesephotoreceptorsaremostly
foundininvertebrateeyes.Incontrast,ciliaryphotoreceptors,foundinvertebrates,employ
phosphodiesteraseandsignaltransductionyieldstothehyperpolarizationofthe
photoreceptormembranepotential(seeNilsson,2004).Cnidariansalsohaveciliarytype
photoreceptors.Sinceboththesecascadesareusedforsignaltransductioninothercelltypes,
itisevidentthatgenesencodingproteinsusedtotransducelightsignalsareevolutionarily
relatedtoothergenes.

Anoculartissuethatincreasesourknowledgeofeyeevolutionisthelens.Lensfocusesthe
lightontheretina.Althoughthelensisnotfoundinthesimplestformoftheeye,itisa
requirementforthecameratypeeye.Therefore,theevolutionaryhistoryofthelensreveals
informationregardingthetypesofimprovementsinvisionnecessaryforcolorvisionand
functionofthevisualcortex.Cameratypeeyesarefoundinevolutionarilydiverse
vertebrates,cephalopodsandcnidarians.Althoughthesethreetypesoflenseshavedifferent
embryologicalorigins(seeTomarevandPiatigorsky,1996),theyappearsimilar.This
characteristicreflectstheirsharedexpressionofwatersolubleproteinsfromthecrystallin
family.Highlevelsofexpressionarerequiredforlenstransparencyanditsrefractiveindex.
Differentmembersofthisfamilymediatelensfunctionintheaforementionedphyla.All
vertebratelensescontainandcrystallins.Twocrystallins,Aand B,arestructurally
andfunctionallyrelatedtoubiquitouslyexpressedsmallheatshockproteins.Sixcrystallins
aredistantlyrelatedtomicrobialstressrelatedproteins(seeWistow,1995).Theyalsoshare
similaritieswithsevencrystallinsandaresometimesreferredtoascrystallins.
Invertebratelensesandsomevertebratelensesalsocontainaspecialclassofenzyme
crystallins.Enzymecrystallinsareidenticalorderivedfrombasicmetabolicenzymes.Avian
genomescontainapairofcrystallingenes.Chicken2crystallinbutnot1crystallinisa
functionalenzymeargininosuccinatelyase(ASL)(seePiatigorsky2003;Wistow1995).
Examplesofadditionalenzymecrystallinsarecrystallin/NADPquinonereductasein
guineapigandcrystallin/aldehydedehydrogenase1inelephantshrew(seeWistow,1995).

Genomicstudiesofcrystallingenesidentifiedgeneduplicationandchangesintheir
regulatorymechanismsaffectingtheirexpressioninsideandoutsideofthelens.Crystallins
servedasthefirstexampleoftheevolutionarystrategy,termedgenerecruitment(see
Piatigorsky2003).Crystallingenerecruitmentbymodifyingthetranscriptionalprogramof
preexistinggenesappearstobeamajormoleculardrivingforceoflensevolution.
Evolutionarystudiesofgenesthatcontrolbraindevelopmentandhigherbrainfunction

furthersupporttheroleofchangesintranscriptionalprogramsduringevolution(see
PichaudandDesplan,2002;Khaitovichetal.2005).

2.Evolutionfromtheprototypeancestoreyetothepresentdaycomplexeyefoundin
vertebrates.

Theissueoftheevolutionaryoriginofthevisualsystemcanbedividedintoatleastthree
phases.Inthefirstphase,weneedtoexplaintheformationofphotosensitivecells(s).Inthe
secondphase,weneedtoaddresstheformationofthemostprimitivevisualsystem.Finally,
theformationoftheprimitiveeye,promptsustoexplainhowthissystemledtothe
developmentofeyesofvaryingcomplexity.Herein,wewillfollowthehypothesisofthe
monophylogeneticoriginofvisualsystemsasproposedbyW.Gehring(seeGehringand
Ikeo,1999;Gehring,2004;2005)andpresentdatathatweakenspolyphyleticmodelsofeye
evolutionproposedearlier(seeNilssonandLand,2002).

ADarwinianprototypeeyethatconsistsofasinglephotoreceptorcellandasurrounding
pigmentcell(seeabove)isactuallyfoundincertainflatwormsandallowsdirectionalvision
(seeGehring,2004;2005).Ithasbeenproposedthatpigmentcellsmayinfactbeevolutionary
precursorstophotoreceptorcells(seeArnheiter,1998).Anintriguingpossibilityexiststhat
pigmentedskincellsperformedthermoregulationandphotoprotectionpriortotheirability
todetectlight.Anancestralanimalwithmelanopsinexpressingpigmentcellsinitsskin
mighthaveevolvedintoanorganismwitharhodopsinexpressingpigmentcellbygene
duplicationandmutagenesisfollowedbyevolutionaryselection.

ToaddressDarwinssproblemwithOrgansofextremePerfectionandComplication,
Gehringarguesthattheevolutionofaneyeprototypewouldseemtobeahighly
improbablestochasticevent,sinceselectioncanonlyworkafterthevariouscomponentsare
assembledintoaprototypethatisatleastpartiallyfunctionalasaphotoreceptororgan(see
Gehring,2004).Hence,thehypothesisofthepolyphyleticoriginoftheeye,arisingasmuch
as65timesindependentlyinhistory(seeNilssonandLand,2002),isincompatiblewiththe
basicconceptofevolutionbynaturalselection.Incontrast,molecularstudies(seenext
paragraphs)arecompatiblewiththemonophylogenicoriginoftheeye.

Finally,animportantissueconcernsthereconciliationoftheevolutionarymodeloftheeye
withfossilrecords.Studiesoffossilssuggestthatthemostactiveperiodineyeevolution
occurredjustpriortoandcontinuingintotheCambrianexplosion(543520millionyearsago)
(seeNilsson,2004).FossilsfromthepreCambrianerashownoobviousrecordsofbilaterians,
butcnidariansappeartohavebeenabundant.Concerningthetimeneededfortheevolution

ofacameralikeeye,atheoreticalmodelbyNilssonandPelgeryieldedtheestimatethatless
than400,000generationsareneededforsuchaprocess.Hence,lessthanhalfamillionof
yearsisthetimeneeded,afractionofthepreCambrianandCambrianeras,togeneratea
complexeyefromitsprototype.

3.Whichgenesregulateeyeformationandhowdothesegenesfunctionindifferent
species?

Identificationofregulatorygenesthatcontroleyedevelopmentcontributessignificantlyto
ourthinkingabouttheevolutionoftheeye.Gehringsargumentaboutthemonophylogenic
originoftheeyeissupportedbyaseriesoffindingsthatevolutionaryconservedgenePax6
controlseyedevelopmentinanumberofvertebratesandinvertebrates.APax6relatedgene,
PaxB,wasfoundincnidariansandwasimplicatedineyeandstatocystdevelopment(see
Kozmik,2005).Inmammalianembryos,Pax6isexpressedineveryimportantoculartissue.
Pax6isexpressedinthesurfaceectodermthatgivesrisetothelensandcornealepithelium.
Pax6isalsoexpressedintheneuroectodermthatformstheneuroretina,retinalpigment
epitheliumandpartsoftheiris.Pax6istransitionallyexpressedinneuralcrestcellsthatform
thecornealendothelium,keratocytes,partsoftheirisandtrabecularmeshwork.However,
Pax6isnotexpressedinvertebratephotoreceptors.InDrosophilacompoundeye,
Pax6/eyeless,isexpressedintheeyeimaginaldisk,anepithelialsheetofcellsthat
differentiateintophotoreceptorsandothercellsoftheadultflyeye.Evidencewasobtained
thateyelessbindstoorthologousregionsofmultipleDrosophilarhodopsinpromotersduring
itsreexpressionphaseinmaturephotoreceptors(Shengetal.1996).

ThefoundationforGehringsevolutionarymodelisthatPax6controlsrhodopsinexpression.
Studiesofcnidarian(jellyfishTripedalia)PaxBindeedhaveshownbindingtotheDrosophila
rhodopsinregulatoryelement(Kozmiketal.2003).Evolutionoftheeyecanbeexplained
usingtheintercalarymechanism(seeGehringandIkedo,1999).Darwinsprototypeofthe
eyehastwogenes,Pax6anditstarget,ancestralrhodopsin.Anincreasinglymorecomplex
eyecanbegeneratedbyintercalationofnovelgenesintothisrelationship.Forexample,Pax6
directlyregulateslenscrystallins(seeCveklandPiatigorsky,1996).Pax6bindingsitesare
showninvertebrateandinvertebratecrystallingenepromotersasshowninFig.1.More
importantly,taxonspecificcrystallinssuchaschicken1andguineapigcrystallin/NADP
quinonereductasewereshowntobedirectlyregulatedbyPax6(seeCveklandPiatigorsky,
1996).AninsertionofagenomicfragmentcontainingPax6bindingsites5ofthecrystallin
promoter(seeWistow,1995)orintothe3rdintronoftheancestorcrystallingeneis
illustratedinFig.2.Itisalsopossibletohypothesizethataregulatoryregioninonegene
encodingancestralsmallheatshockproteinwasconvesrtedintoapax6bindingsitesbyjust
threenucleotidesubstitutions(seeFig.3).Therecruitmentsofenzymecrystallinsare

relativelyrecentevolutionaryeventstolensfunctioninspecificanimallineagesandprovide
directevidenceforintercalaryevolution.

Furthermore,Pax6regulatestheexpressionofspecializedregulatorygenesrequiredforeye
formationinthemouseandfly.Themostdocumentedexamplesarehomeoboxcontaining
genes,Six3/sineoculis(so),andstructurallyunclassifiedgenes,Mab21,thatareregulatedby
Pax6inthesetwodiversesystems(seeLang,2004;Michautetal.2003).Thus,currentdata
showthatthekeyregulatorygeneforeyedevelopmentPax6/eyelessregulatesexpressionof
manyregulatoryandstructuralproteinsrequiredforvision.Newdatafrommolecular
studiesofvisualsystemdevelopmentinbothvertebratesandinvertebratescombinedwith
analysisofgenomesfromthesespeciesandspeciesthatdonothavevisualsystemswill
generateadditionaldatatofurtherourunderstandingoftheevolutionofvision.

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2006

Figure 1. A diagrammatic summary of promoters encoding vertebrate and invertebrate crystallins.

Maf responsive element, MARE; retinoic acid responsive elements, RARE; and cAMP responsive
elements, CRE are shown in orange, light yellow and yellow, respectively. Note that all these promoter
contain Pax- and large Maf-binding sites (see Cvekl et al. 2004).

Figure 2. Diagram of -, - and -crystallin genes (adopted from Wistow, 1993;1995). -crysallin
locus is transcribed from two promoters. The proximal, lens specific promoter contains MARE/Pax6binding sites. Avian -crystallin loci contain insertion of a lens-specific enhancer in the third intron.
This insertion is absent in mammalian genomes.
Duck -crystallin locus has TATA-box in the promoter like the majority of crystallin promoters. Pax6binding sites in this locus remain to be studied.

Figure 3. A model to explain conversion of ancestral small heat shock protein gene promoter into a
Pax6-responsive promoter. Note that a strong Pax6-binding site contains motifs recognized by heat
shock transcription factors and AP-1 proteins.