Drosophila

development
Suman Bhattacharjee
Drosophila life cycle
Embryology
overview
Development
al
Progression :
Gene
interplay
 Anterior and posterior
system

Bicoid
Nanos
Anterior system by Bicoid gen
Posterior system by nanos, caudal and Oskar
gene
Terminal axis determination by Torso
Dorso-ventral system - ventral signal
Dorso-ventral system - ventral signal
Dorsal signaling by
Gurken and
Torpedo
Microtubule rearrangement : A key player
Determining initial polarity by
interaction with the follicle cells

Towards anterior Towards posterior Towards dorsal

Gastrulation
Germ band
extension
Parasegm
ents
Gastrulation - organiser
Para-segments produce
Imaginal discs
A thickening of the epidermis of an
insect larva which, on pupation, develops
into a particular organ of the adult insect.

9 pairs (18) + 1 fused pair

Predetermined for developmental fate
Epidermis ------------------ Invaginates
Thickening. (Partly folded)
Body
segmentation
proteins
Movement of mRNA and
proteins change from the
oocyte to the Embryo
Body segmentation
 Genes Affecting Body
Segmentation
Products of Maternal Bicoid, Nanos, Oscar
Effect Genes establish
gradients in the egg

Gap genes define

broad areas and
regulate Pair-rule
genes

Pair-rule genes refine

segment locations and
regulate Segment Polarity
genes

Segment Polarity
genes determine Homeotic genes
segment boundaries define the role of
and orientations each segment
Development
al
Progression
SEGMENTATION IN
DROSOPHILA
Maternal Effect Genes

Control development
Segmentation Genes of Segments

Homeotic Genes
WHAT HAVE WE LEARNT SO FAR
 SEGMENTS OF A DROSOPHILA
All arthropods are segmented. The body of Drosophila melanogaster is built from 14 segments.

 3 segments make up the head with its antennae and mouth parts.
 3 segments make up the thorax. Each thoracic segment has a pair of legs. In Drosophila (and other flies), the
middle thoracic segment carries a single pair of wings; the hind segment a pair of halteres.
 8 abdominal segments.
 SEGMENTATION GENES – Formation of
‘Molecular Blueprint’

Gap
Genes

Pair Rule
Genes

Segment
Polarity
Genes

Segmentation is a stepwise exercise that divides the embryo up into ever smaller units. Its like for cutting a cake into a large
number of equal slices, you would first cut it into large chunks then progressively cut each chunk into smaller slices.
MUTANTS OF SEGMENTATION GENES
 Examples
Gap genes Krüppel (Kr) Pair-rule genes fushi tarazu (ftz)
Secondary
knirps (kni) odd-paired (opa)
hunchback (hb) odd-skipped (slp)
giant (gt) sloppy-paired
(slp)
tailless (tll) paired (prd)
huckendein (hkb)
buttonhead (btd) Segment engrailed (en)
empty spiracles polarity genes wingless (wg)
(ems)
hedgehog (hh)
Pair-rule genes hairy (h) fused (fu)
Primary
even-skipped armadillo (arm)
(eve)
runt (run) patched (ptc)
gooseberry (gsb)
 PAIR RULE GENES
First Indication of Segmentation
 The transcription patterns of these genes are striking in that they divide the embryo into the areas that are
the precursors of the segmental body plan

 One vertical band of nuclei (the cells are just beginning to form) expresses a pair-rule gene, then another
band of nuclei does not express it, and then another band of nuclei expresses it again. The result is a
“zebra stripe” pattern

 How are some nuclei of the Drosophila embryo told to transcribe a particular gene while their neighbors
are told not to transcribe it? The answer appears to come from the distribution of the protein products of
the gap genes

 Three genes are known to be the primary pair-rule genes — hairy, even-skipped, and runt—are essential
for the formation of the periodic pattern, and they are directly controlled by the gap gene proteins. The
enhancers of the primary pair-rule genes are recognized by gap gene proteins, and it is thought that the
different concentrations of gap gene proteins determine whether a pair-rule gene is transcribed or
not
PERIODIC EXPRESSION OF PAIR RULE GENE
even-skipped

eve ftz
 EXPRESSION OF SEGMENT POLARITY GENES
 Once cells form, interactions take place between the cells which are
mediated by the segment polarity genes

 Through this cell-to-cell signaling, cell fates are established within

each parasegment.

 Encoded proteins are constituents of the Wingless and Hedgehog

signal transduction pathways
 One row of cells in each parasegment is permitted to express the
Hedgehog protein, while the other expresses the Wingless protein

 Activation of Engrailed gene - cells express Hedgehog protein

 In turn, engrailed gene is activated when cells have high levels of the
Even-skipped or Fushi tarazu transcription factors
 Even skipped (Fushi tarazu) -> engrailed -> Hedgehog = anterior
 Engrailed transcription marks the anterior boundary of each
parasegment
 The wingless gene is activated – presence of Sloppy-paired protein.
Marks posterior boundary
 Sloppy-paired protein -> wingless = Posterior
Unique expression of genes in different parasegments of Drosophila embryo
Anterior and posterior
boundary determination
in each body segments
EXPRESSION OF Engrailed GENE
 Metamorphosis –
Molecular approach
OUTLINE
• Introduction
• Broad-complex transcription factors
• Signalling of metamorphosis by Ecdysone
• Signalling of metamorphosis by Juvenile hormone (JH)
• conclusion
• Insect metamorphosis can be classified into three modalities:
• ametabolan (no changes),
• hemimetabolan (progressive changes) and
• holometabolan (dramatical changes at the end of the cycle).

• The metamorphic changes are mainly regulated by two hormones: the

moulting hormone, which promotes the moults, and the juvenile
hormone (JH), which represses the transformation into the adult..

• The action of these two hormones is mediated by a number of

transcription factors, and the molecular mechanisms regulating the
expression of these and of the corresponding target genes are finally
refined by the action of micro ribonucleic acids (miRNA)
 Ecdysone and Juvenile hormone interplay
• Ecdysone: Pulses of 20-hydroxyecdysone initiate each of the major
developmental transitions, including both larval molting and
metamorphosis
• Juvenile hormone (JH) prevent precocious metamorphosis allow the
larva to undergo multiple rounds of molting until it attains the proper
size for metamorphosis
• JH is classically viewed as an antimetamorphic hormone a high titer of
JH in the early larval instars directs ecdysone to initiate molting, while
the absence of JH during the final instar allows ecdysone to trigger the
morphological changes of metamorphosis

Metamorphosis
Molting Ecodysone
Juvenile (JH)
Broad-complex transcription factors
• Important group of
transcription factors
involved in metamorphosis
is that known as Broad-
Complex (BR-C). The BR-C
family of transcription factors is
especially interesting because
they have different ways of
action in hemimetabolans and
holometabolans.

• Functional studies in Drosophila

melanogaster and the
lepidopterans Manduca sexta
and Bombyx mori have shown
that BR-C factors are essential
for the transformation of last
instar larvae into pupae
 Role of Ecdysone
• Ecdysone is a direct regulator of transcription and, like other steroid
hormones acts through a nuclear receptor pathway

• The ecdysone receptor is heterodimer of the nuclear receptors EcR and

USP that binds to the ecdysone response element (EcRE)
• Ecodysone (EcRE) -> ecdysone receptor and USP -> activates
Transcription factors (HR3, HR4, HR39, E75, E78, FTZ-F1, etc.)

• Upon binding to their heterodimeric receptor composed by two nuclear

receptors, the ecdysone receptor (EcR) and the ultraspiracle (USP)
activate the expression of a hierarchy of transcription factors (HR3, HR4,
HR39, E75, E78, FTZ-F1, etc.) that regulate the expression of the target
genes underlying the cellular changes associated to molting and
metamorphosis
 Function of JH
• The multifunctional nature of JH has impeded the characterization of
its signaling mechanisms.

• JH is pleiotropic and controls not only development but also

sexual behaviour, pheromone production, caste determination,
diapause, migration, and the synthesis of female yolk proteins and
male accessory gland proteins

• In regulating these diverse processes JH appears to utilize multiple

pathways, some of which involve the activation of gene expression
while others are transcription-independent
 JH receptor mutation
• Activity of JH in many insects influences larval development so
mutation of JH receptor would have severe consequences for larval
development
• In Drosophila, JH is essential to pre-adult development as its removal
by genetic ablation of the corpora allata results in lethality around
the time of head development and defects in larval development
including precocious apoptosis of the fat body

Methoprene-tolerant (MET) is
identified as a potential JH receptor
This was confirmed in the red flour beetle
Tribolium castaneum, where removal of
Met expression produces a premature
and lethal initiation of pupation
Metamorphosis is regulated by Hormones and environmental factors
Flower development in
Arabidopsis
 Flower development in Arabidopsis

1. Flowering time gene or heterochrony – conversion from vegetative meristem to floral meristem
2. Flower meristem identity gene – Regulates the formation of flower
3. Cadastral gene – Whorl formation in flower
4. Homeotic genes – Regulates structure and position of whorl
Flower development in Arabidopsis
Autonomous and conditional
specification
Cell specification : Autonomous,
conditional and syncytial
specification
• Autonomous specification results from cell-intrinsic properties , which arise from a
cleavage of a cell with asymmetric cytoplasmic determinants or morphogenetic
determinants.
• Thus, the fate of the cell depends on factors segregated into the cytoplasm during
cleavage. Autonomous specification happens right from the beginning, when egg
cytoplasm is partitioned asymmetrically. It is interesting to note that this contribution
is entirely maternal , since the sperm donates only a negligible amount of cytoplasm.
• Conditional specification happens by cell-extrinsic cues such as cell-cell contact and
temporal-spatial signaling (wnt, notch, fgf, bmp, hedgehog, etc). This depends a lot on
the relative position of the cell in the embryo.
• . Syncytial specification - This type of a specification is a hybrid of the autonomous
and conditional that occurs in insects. This method involves the action of morphogen
gradients within the syncytium. As there are no cell boundaries in the syncytium, these
morphogens can influence nuclei in a concentration-dependent manner.
Autonomous and conditional specification
 One example
• Ectoderm, and more specifically, neuroepithelium is widely regarded
as the default program in vertebrate development . That is, in the
absence of any other differentiating signal, a stem cell will proceed
towards neuroepithelial fate.
Spemann Mangold experiment : Organizer concept
 Spemann Mangold experiment : The result

• The remarkable results:

• the transplanted tissue developed into a second
notochord
• neural folds developed above the extra notochord
• these went on to form a second central nervous system
(portions of brain and spinal cord) and eventually
• a two-headed tadpole.
 Spemann Mangold experiment : The mechanism
• Cells on the ventral side of the blastula secrete a variety of
proteins such as bone morphogenetic protein-4 (BMP-4)
• These induce the ectoderm above to become epidermis.
• If their action is blocked, the ectodermal cells are allowed
to follow their default pathway, which is to become nerve
tissue of the brain and spinal cord.
• The Spemann organizer blocks the action of BMP-4 by
secreting molecules of the proteins
• chordin and
• noggin
• Both of these physically bind to BMP-4 molecules in the
extracellular space and thus prevent BMP-4 from binding
to receptors on the surface of the overlying ectoderm cells.
• This allows the ectodermal cells to follow their intrinsic
path to forming neural folds and, eventually, the brain and
spinal cord.
 Spemann Mangold experiment : The explanation

• In the Spemann/Mangold experiment,

transplanting an organizer to the
ventral side provided a second source
of chordin. This blocked BMP-4
binding to the overlying ectoderm and
thus changed the fate of those cells to
forming a second central nervous
system rather than skin.
Hensen’s Node :
What is it ?
Hensen’s Node : Maturation of
Embryo
Hensen’s Node : Functional role
 Nieuwkoop center : What is
it ?
• Dorsal most vegetal most region

The Nieuwkoop Center is the dorsal- and

vegetal-most cell of the early blastula. It gives rise
to the Primary Organizer, which is the dorsal lip
of the blastopore (DLB). The Primary Organizer has
a dorsalizing effect, and together with the Sperm
Entry Point (SEP) gives rise to the dorsal/ventral
axis. Dorsalized tissue gives rise to somites and
pronephric tubules.
 Nieuwkoop center : Functional Role

Lateral and ventral  Dorsal

Ectoderm  Neural Tissue
and brain
Limb Development in
vertebrates
AER – Apical ectodermal ridge (AER)
ZPA - zone of polarizing activity
(area of mesenchyme)
Limb Development : Overview
AER – Apical ectodermal ridge (AER
ZPA - zone of polarizing activity
(area of mesenchyme)
FGF Cell signaling
The Mechanism