How do cells become different

from one another?

Objectives
1. Findouthowcellscantelleachotherapart;cell
fatedecisions.
2. HowDrosophiladeterminesanteriorposterior
3. Findoutwhatisamaternalmutantsanda
maternalscreenare.
4. Whatisnecessaryandwhatissufficient.
5. HowisaMorphogen
6. HowisaGradient

Drosophila malenogaster

Drosophila malenogaster

FlyAnatomy
Egg

Anterior

Posterior

FirstInstarLarva

Removeanteriorpolecytoplasm?

Conclusions?

Conclusions?

TheAxisisEstablishedUnder
MaternalControl

MaternalEffectMutations
MutationsthatdoNOTresultindamagetothe
motherbuthaveeffectsonheroffspringthatcan
notberescuedbywildtypesperm.

MaternalEffectScreen

Nophenotype
becauseitis
recuedby
mothers+allele

Whatexperimentwilltellyou
Bicoidisnecessaryforthe
anterior?

MutationsAffectingthe
AnteriorPosteriorAxis

BcdisRequiredfortheFormationof
AnteriorStructures

PhenocopiesRemovalofAnteriorCytoplasm

Whataretheexperimentsto
showBicoidissufficient?

TransplantationofWildtypeAnteriorCytoplasm
CanRescueBcdMutants

QuantitativeRelationshipBetweentheNumber
ofBicoidGenesandthePatternoftheEmbryo

From:TheMakingofa
Fly,Lawrence

Hypothesis:Bcdestablishesthe
anteriorposterioraxisthroughthe
establishmentofamorphogen
gradient.

TransferofBcdRNAisSufficienttoInduce
AnteriorStructures

BcdMayActasaMorphogen

Morphogen
Aninducingfactorthatcanevokemore
thanonecellstatefromtheresponding
tissue.

TransferofBcdRNAisSufficienttoInduce
AnteriorStructures

BcdMayActasaMorphogen

Gradient
Theasymmetricdistribution
ofaproteinorprotein
activityinatissue.

TransferofBcdRNAisSufficienttoInduce
AnteriorStructures

BcdMayActasaMorphogen

Hypothesis:Bcdestablishesthe
anteriorposterioraxisthroughthe
establishmentofamorphogen
gradient.
Prediction:Bcdmustbea
cytoplasmicdeterminantlocalized
totheanteriorpole.

HowdowetestthisPrediction:
1. AttheRNAlevel?
2. Attheproteinlevel?

AttheRNAlevel?
Recitationthisweek
Drawadiagramofhowtoconductawholemountinsitu
andincludethefollowing:
1.Aschematicofthebcdgenelabeling5primeend,3
primeend.
2.TheresultingRNAprobelabeling5primeend,3prime
end.
3.Alistofpositiveandnegativecontrols.
4.Drawthedrospholiaeggcoloringintheresultofthebcd
probeandalllistedpositiveandnegativecontrolsand
statewhythecontrolsgaveyourpredictedpattern,

Attheproteinlevel?
1.Howwouldyoudothis?
2.Whatwouldyouuseaspositiveandnegativecontrols
andwhataretheycontrolling?
3.Predictedresultsincludingthatofcontrols.