Looking for the appropriate size:

genetics under control

Crazy about Biomedicine May 2013

Ana Ferreira
Development and Growth Control Lab
 Summary
I. Genetics

Definition
Mendelian Genetics

Drosophila melanogaster: The Fruit Fly

Historical view of the fly

Drosophila as a model organism

II. Developmental Biology

Definition
Historial view

III. Growth Control:

The different parameters

Our system: the fly wing
Systemic vs Organ-autonomous Growth Control
Size Control and Human Disease
I. Genetics
 Genetics

is a discipline of biology, is the science of genes, heredity, and

variation in living organisms

Genetics deals with the molecular structure and function of

genes, gene behavior in the context of a cell or organism, patterns
of inheritance from parent to offspring, and gene distribution,
variation and change in populations

GENETICS + ORGANISM EXPERIENCES

=
FINAL OUTCOME
Mendelian and Classic Genetics

Gregor Mendel
(1822 - 1884)

studied the nature of inheritance in plants

observed that organisms inherit traits by way

of discrete units of inheritance, which are now
called genes

traced the inheritance patterns of certain

traits in plants and described them
mathematically
 Discrete Inheritance and Mendels Laws

studied the segregation of heritable traits in pea plants

29,000 pea plants

Grow easily, develop pure-bred strains, and control their pollination

Pisum sativum
Discrete Inheritance and Mendels Laws
 Discrete Inheritance and Mendels Laws

Dominant trait
Alleles: is one of a number of alternative forms of the same gene
Discrete Inheritance and Mendels Laws
 Discrete Inheritance and Mendels Laws

3:1 ratio

diploid species: each individual has two copies of each gene, one inherited from each parent

organisms with two copies of the same allele

of a given gene are called homozygous

organisms with two different alleles of a given gene are called

heterozygous
Discrete Inheritance and Mendels Laws

homozygous heterozygous homozygous

(WW) (Ww) (ww)

Purple Purple White

 Discrete Inheritance and Mendels Laws

homozygous heterozygous homozygous

Genotype (WW) (Ww) (ww)

(set of alleles)

Phenotype Purple Purple White

(observable traits)

W W
one allele is called other allele is called
dominant recessive
Discrete Inheritance and Mendels Laws
Discrete Inheritance and Mendels Laws
Discrete Inheritance and Mendels Laws

3:1 ratio
Discrete Inheritance and Mendels Laws
 Discrete Inheritance and Mendels Laws

1 The Law of Dominance: In a cross between contrasting homozygous

individuals, only one form of the trait will appear in the F1 generation -
this trait is the dominant trait
 Discrete Inheritance and Mendels Laws

1 The Law of Dominance: In a cross between contrasting homozygous

individuals, only one form of the trait will appear in the F1 generation -
this trait is the dominant trait

2 The Law of Segregation: when any individual produces gametes, the

copies of a gene separate so that each gamete receives only one copy
(allele) - a gamete will receive one allele or the other
 Discrete Inheritance and Mendels Laws

1 The Law of Dominance: In a cross between contrasting homozygous

individuals, only one form of the trait will appear in the F1 generation -
this trait is the dominant trait

2 The Law of Segregation: when any individual produces gametes, the

copies of a gene separate so that each gamete receives only one copy
(allele) - a gamete will receive one allele or the other

3 The Law of Independent Assortment: alleles responsible for different

traits are distributed to gametes (and thus the offspring) independently
of each other
Drosophila melanogaster
Drosophila melanogaster: the fruit fly
Drosophila melanogaster: the fruit fly
Historical view of Drosophila

Charles W. Woodworth
(1865 - 1940)

1900 First to breed Drosophila in the Lab

Historical view of Drosophila

Thomas Hunt Morgan (1866 - 1945)

1900 Started to work with Drosophila
(study of mutation)

1910 First mutation was found (white)

1911 Genes are on chromosomes

1933 Nobel Prize in Physiology or Medicine

for the role played by chromosomes in
heredity
Historical view of Drosophila
Historical view of Drosophila

Hermann Joseph Mller

(1890 - 1967)

1946 Nobel Prize in Physiology or Medicine

for the discovery of the genetics effects
of Radiation (X-ray mutagenesis)
 Historical view of Drosophila

Eric Wieschaus Janni Nusslein-Volhard Edward B. Lewis

(1947 - ) (1942 - ) (1918 - 2004)

1995 Nobel Prize in Physiology or Medicine for revealing

the genetic control of embryonic development
 Historical view of Drosophila

Jules A. Hoffmann
(1941 - )

Bruce A. Beutler Ralph M. Steinman

(1957 - ) (1943 2011)

2011 Nobel Prize in Physiology or Medicine for the discovery

of the dendritic cell and its role in adaptive immunity
Why Drosophila melanogaster is such a good model organism ?
Why Drosophila melanogaster is such a good model organism ?

Short Life Cycle (Temperature Dependent 10 days @ 25C)

Each Female lays 400-500 eggs

Easy to maintain in the Lab (low cost)

Suitable of Genetic Manipulation

Extensive set of genetic tools available

Simple karyotype: 4 pairs of chromosomes (3 autosomes + sexual chromosomes)

Gene Sequence Conservation with humans: 60%

Functional conservation of regulatory and biochemical pathways with humans

Why Drosophila melanogaster is such a good model organism ?

Short Life Cycle (Temperature Dependent 10 days @ 25C)

Each Female lays 400-500 eggs

Easy to maintain and manipulate in the Lab (low cost)

Suitable of Genetic Manipulation

Extensive set of genetic tools available

Simple karyotype: 4 pairs of chromosomes (3 autosomes + sexual chromosomes)

Gene Sequence Conservation with humans: 60%

Functional conservation of regulatory and biochemical pathways with humans

Why Drosophila melanogaster is such a good model organism ?

Short Life Cycle (Temperature Dependent 10 days @ 25C)

Each Female lays 400-500 eggs

Easy to maintain and manipulate in the Lab (low cost)

Suitable of Genetic Manipulation

Extensive set of genetic tools available

Simple karyotype: 4 pairs of chromosomes (3 autosomes + sexual chromosomes)

Gene Sequence Conservation with humans: 60%

Functional conservation of regulatory and biochemical pathways with humans

Why Drosophila melanogaster is such a good model organism ?

Short Life Cycle (Temperature Dependent 10 days @ 25C)

Each Female lays 400-500 eggs

Easy to maintain and manipulate in the Lab (low cost)

Suitable of Genetic Manipulation

Extensive set of genetic tools available

Simple karyotype: 4 pairs of chromosomes (3 autosomes + sexual chromosomes)

Gene Sequence Conservation with humans: 60%

Functional conservation of regulatory and biochemical pathways with humans

Why Drosophila melanogaster is such a good model organism ?

Short Life Cycle (Temperature Dependent 10 days @ 25C)

Each Female lays 400-500 eggs

Easy to maintain and manipulate in the Lab (low cost)

Suitable of Genetic Manipulation

Extensive set of genetic tools available

Simple karyotype: 4 pairs of large chromosomes (3 autosomes + sexual chromosomes)

Gene Sequence Conservation with humans: 60%

Functional conservation of regulatory and biochemical pathways with humans

Why Drosophila melanogaster is such a good model organism ?

Short Life Cycle (Temperature Dependent 10 days @ 25C)

Each Female lays 400-500 eggs

Easy to maintain and manipulate in the Lab (low cost)

Suitable of Genetic Manipulation

Extensive set of genetic tools available

Simple karyotype: 4 pairs of large chromosomes (3 autosomes + sexual chromosomes)

Gene Sequence Conservation with humans: 60%

Functional conservation of regulatory and biochemical pathways with humans

Why Drosophila melanogaster is such a good model organism ?
Drosophila melanogaster Life Cycle

Growth Phase
Drosophila melanogaster: why is such a potent genetic organism ?

Genome fully sequenced

Mutant animals are readily obtainable

Huge amount of transgenic lines available

Targeting gene expression in a temporal and spatial fashion

Targeting gene expression: Gal4-UAS System

Driver line Responder line

Big collection of both Driver and Responder Lines available

Temperature Dependence of the Driver Line
Targeting gene expression: Gal4-UAS System
Targeting gene expression: Gal4-UAS System
II. Developmental Biology
Developmental Biology
 Historical Perspective The first steps

Aristotle
(384 322 AC)
Study of the Development of the chick

The semen of the male provides the form or

soul and the female the unorganized matter
(menstrual blood) allowing the embryo to
grow: EPIGENESIS

Theory of Preformationism: organs with their own

shape expand

Theory of Spontaneous Generation: life of

invertebrates emerges from non-living matter
(nothing)
 Historical Perspective - Renaissance

Leonardo da Vinci
(1452 - 1519) Dissection of human corpses

Drawings of the vascular and system

First drawing of the human fetus in the

utero

Views of a Fetus in the Womb

Leonardo da Vinci, ca. 1510-1512
Historical Perspective - Renaissance
 Historical Perspective - Renaissance

Antonie van Leeuwenhoek

(1632 - 1723)
Discovered the microorganisms: animacules

Discovered the spermatozoa

now that I have discovered that the

animalcules also occur in the male seed of
quadrupeds, birds and fishes, I assume with
even greater certainty than before that a
human being originates not from an egg but
from an animalcule that is found in the male
semen
 Historical Perspective - Renaissance

PREFORMATIONISM

organisms develop from

miniature versions of themselves

Nicolaas Hartsoeker in 1695

 Historical Perspective - Renaissance

Reiner de Graaf
(1641 - 1673)

Discovered the follicles of the ovary (known as

Graafian follicles), in which the individual egg
cells are formed

Rejecting the preformationism

 Historical Perspective

Ernst Haeckel
"ontogeny recapitulates phylogeny (1834 - 1919)

Recapitulation Theory /
Embryological Parallelism

developing from embryo to

adult, animals go through
stages resembling or
representing successive
stages in the evolution of
their remote ancestors
Historical Perspective

Karl Ernst von Baer

(1792 - 1876)

Opposing view that the early general forms

diverged into four major groups of specialized
forms without ever resembling the adult of
another species
 Historical Perspective

August Weismann Germ plasm theory

(1834 - 1914)
inheritance only takes place by means of the germ
cellsthe gametes

Other cells of the bodysomatic cellsdo not

function as agents of heredity
 Historical Perspective

Experimental Embryology

Wilhelm Roux
1888 Experiment destroying the frog embryo (in the two cells stage)

Hans Driesch
1892 Separates de early 4 cells stage embryo of the sea urchin

Hans Spemann and Hilde Mangold

1918-1924 Transplants of cells from one embryo to another induced particular
tissues or organs embryonic induction. Nobel Prize in 1935
Are Developmental Biology and Genetic Linked ?
III. Growth Control
How are differences in size achieved ?
 What determines differences in size ?

Size of an organ/animal = number of cells + size of the cells + space between cells

Size of an organ/animal = number of cells + size of the cells similar

Cell Number Cell Division

+
Cell Death

Cell Number
+
Cell Size
Cell Growth
Cell Size
 What determines differences in size ?

Cell Division / Proliferation: increase in cell number by one cell (the

"mother cell") dividing to produce two "daughter cells"

Cell Death / Apoptosis: is death of a cell in any form, mediated by an

intracellular program (DNA fragmentation and protein degradation)

Cell Growth: increase in cell mass (protein synthesis and organelle

biogenesis) Cell Cycle
How organs achieve a particular size and pattern ?
Drosophila imaginal discs: proliferative tissues
 Drosophila wing imaginal disc

Embryo 20-30 cells

Larvae

wing
Adult

50,000 cells
notum
Drosophila wing imaginal disc development
Body Size Regulation
 Systemic vs organ-autonomous growth control

Long range signaling molecules Cell autonomous growth promoters

(hormones)
Environmental factors Morphogens, signaling molecules
(nutrition)
Systemic growth control

SYSTEMIC GROWTH CONTROL

GROWTH RATE DEVELOPMENTAL TIMING

(moults+pupariation)
 Systemic growth control

DEVELOPMENTAL TIMING

Ecdysone

Ring gland

Fat body
Insulin
Brain
nutrients Gut

Hemolymph (fly blood)

FEEDING
GROWTH
 Organ-autonomous growth control

Transplants Experiments: when a small organ is transplanted into an adult

organism it grows to its normal final size (even in between different species)

Regeneration Experiments
 Size Control and Human Disease

Cancer: Organ hypertrophy

tumor initiation, or atrophy
metastasis

Growth Pathways
Insulin pathway
dMyc oncogene
Hippo pathway
TGFb signaling (Dpp)
Wnt signaling (Wg)

Diabetes Regeneration
and and Stem
Obesity Cell Biology

Drosophila was, is and will be important for Human Biology

 Thank you

Development and Growth Control Lab

Crazy about
B omedicine
Transformation in flies