Patterning Early Dm Embryo

Patterning AP axis: Gap gene expressions

Establishment of parasegments
Readings: Chapter 2 Wolper, pp52-65
Muhammad Tariq

2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

HOW Dpp gradient is established from uniform Dpp levels:

2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

Patterning AP axis: gap gene expressions

Expression of gap genes divides AP into broad regions

Embryo is still acellular when these first zygotic genes are active
All code for txn factors

Initiation of gap gene expression

bicoid gradient in AP axis

bicoid..........hb.........giant, krppel, knirps, expressed in sequence

Gap gene proteins and mutants

mutants miss large body sections

short lived proteins anf half life in minutes

2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

Relationship between bicoid and hb

Bicoid provides positional signal for anterior exp of hb
Zygotic expression of hb- anterior half of embryo
Zygotic expression superimposes above maternal hb
hb suppressed by nanos in posterior end

2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

Hb gradient and other gap genes: txn factors

Activation and repression

hb txn factor: acts as a morphogen

krppel activated by bicoid and low hb, repressed at high hb
below threshold krppel is not active, so a precise and
localized krppel band

Dose dependent expression of Kr

bicoid- only maternal hb....kr is even more anterior

Overlapping regions of gap genes:

cross inhibition
anterior border of Kr 4-5 nuclei posterior to nuclei that
express giant

A-P axis: number of unique regions on the

Basis of overlapping and graded distribution
of different txn factors
2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

How to analyze this relationship, how to determine Hb gradient?

experiment

2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

Establishment of parasegments: activation of pair rule genes in A-P axis

First visible sign of segments: transient grooves on surface of embryo after gastrulation
these grooves are actually parasegments, total 14 PS
ps is fundamental unit of segmentation in Dm

Segment = posterior region of one PS and

anterior of next
PS in thorax and abdomen = delimited by activity
of pair rule genes
Each of pair rule gene is expressed in a series of
Seven transverse stripes along embryo
Stripes in alternate PS
Even skipped define odd numbers
Ftz defines even numbered ps
2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

Striped gene expression pattern present before cellularization

Eve: odd segments
Ftz: even segments

Eve: anterior margin of stripe corresponds to anterior boundary of parasegment

Domains of other pair-rule genes may cross parasegment boundaries
2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

Gap gene activity positions stripes of pair rule genes

Striped expression patterns appear gradually: initially Eve is fuzzy but eventually
acquires a sharp anterior margin

Each stripe is specified independently: e.g 2nd stripe of eve

2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

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Gap gene activity positions stripes of pair rule genes

Normal expression iof bicoid, and three gap genes specify second eve stripe
bicoid & bb: activate eve however dont define the boundaries
kruppel, giant specify boundaries based on repression of eve

Each cis-regulatory site controls expression in a different stripe:

Q: How would you determine it experimentally?

2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

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Establishment of Segmentations and Compartments:

Gap genes

pair-rule genes

segmentation genes

Pair rule genes define anterior boundaries of all 14 ps

Since pair-rule genes like gap genes are transient, how ps boundaries are fixed?
Segmentation genes: unlike gap and pair rule gene, diverse group of genes
No obvious relationship to each other in their protein products
Expressed in all 14 transverse stripes in response to pair rule genes

Pair rule genes act in cellular environment rather than in blastoderm?

Why?
Engrailed (En). Segmentation gene expressed in anterior region of every ps
En is a selector gene
2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

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Engrailed (En): unlike gap and pair rule genes en expressed throughout fly life
Engrailed (En): first appears as 14 stripes at cellularization
Engrailed (En) expression controlled by pair rule genes
Ftz: en is missing in even numbered ps
Inject fluorescent compound in
embryo
Activate single cell witth UV beam

3 cells layer
Anterior region of a ps defines boundary of a cell-lineage restriction
Cells and their descendants from ps never move into adjacent one: genetic
control
Domains of lineage restriction: compartment
Contains all the descendants of cells present when compartment is set
2/11/2014
BIO 315 - Dev. Bio.; Muhammad
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up
Tariq; Lecture 7

Cell-lineage restriction of anterior region of ps carried over to segments

Segment: divided into anterior and posterior compartments

2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

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Genetically mosaic flies: tool to visualize compartment boundaries

Minute technique: cells divide faster, much more
Wing clones
10 div in imaginal discs after blastoderm

No structural features dictate

compartment boundar
Patterns of wing is not
dependent on cell lineage

en: no boundary left

posterior to anterior transform

2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

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Segmentation genes stabilize ps boundaries:

Seg. genes set up a focus of signaling at the boundary that patterns segment
Larva: each segment has well defined anterior and posterior pattern seen on ventral
Epidermis of abdomen
Anterior region has denticles, posterior region is naked
Each segment has unique pattern

segmentation genes alter dentical patterns

e.g.
wg- whole ventral abdomen covered with denticles
anterior region of each segment is duplicated in mirror image
hh- also similar phenotype

2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

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Denticle patterns depend on correct establishment and maintenance

of embryonic ps boundaries

Establishment of ps boundary: Wg, Hh, En involvement

Sometime after ps boundary has been established, a deep groove develops at the
posterior edge of each en expressing stripe and marks edge of each segment
2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

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Interaction between En,

Wg and Hh at compartment
boundaries
Interaction divides continuous epithelium
of abdomen into a succession of anterior
and posterior compartments

Wg moves posteriorly over the

compartment boundar as well as moves
anteriorly to the cells where expressed
Posterior move: short distance as
degraded than anterior distance
Wg repress genes for cuticle formation

2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

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Patterning within compartment boundaries

Epidermis of each abdominal segment is subdivided into a number of type: at least 5
in anterior and 3 in the posterior compartment

Bristles, pigmentation and gene expression

2/11/2014

BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

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BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

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BIO 315 - Dev. Bio.; Muhammad

Tariq; Lecture 7

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