DROSOPHILA MELANOGASTER

A MODEL ORGANISM FOR RESEARCH

 CLASSIFICATION
Kingdom : Animalia
Phylum :Arthropoda
Class : Insecta
Order : Diptera
Family : Drosophilidae
Genus : Drosophila
Species : Melanogaster

HABITAT
 Found in almost all the temperate regions of the world.
 Commonly found in vineyards, trash heaps all around the world, and
 MORPHOLOGY
 Color : Yellowish in color with several black bands across
the abdomen.
 Size : Small about ⅛ inch in length.
 Sexual Dimorphic species ( male and female can be easily
distinguished on the basis of several morphological differences).
 As in other insects, there are 3 main body parts ( tagmata ):
Head , Thorax and Abdomen
 HEAD, THORAX AND ABDOMEN

Head is movable and has the following sensory Organs:

1) Vision : Compound eyes and ocelli
2) Olfactory: Antennae , maxillary palps
3) Gustation : Proboscis
4) Hearing : Antennae
5) Touch : Numerous mechano sensory bristle
 A pair of eyes are located on the external head capsule.
 Each compound eye contains 750 units called Ommatidia.
Primary pigment cells of wild type flies have reddish pigment.
 Ocelli is found on the dorsal aspect of head as triplet – 2 lateral and 1 medial together forms triangle.
 Ocelli and compound eye provides two anatomically separate & functionally different visual pathways.
ANTENNAE
 A pair of movable antennae is located between the compound eyes.
 Antennae is made up of 6 segments/antennomeres :
Single segment of scapel
Two segments of Pedicel
Three segments of flagellum
 Antennae serves as a sensory organ.
MOUTH (Sucking and Sponging type)
 Mouth of Drosophila consist of :
Maxillary palpa: located ventral to the antennae
contains olfactory receptor neuron
Proboscis : Adult fly’s feeding appendages.
Labella : involved in food detection and intake.
Labrum : Food canal, through which liquids are pumped into
the gut.

THORAX
Consist of 3 parts :
A) Prothorax : Anterior , carry one pair of legs.
B) Mesothorax : Middle, carry one pair of leg and one pair of wings.
C) Metathorax : Posterior, carry one pair of legs and one pair of halters
( modified wings ).
WINGS
Wings contain wing cells and wing veins

HALTERES
 Reduced hindwing and acts as a balancing organ.
 Consists of 3 parts scabellum , pedicel and capitellum.

LEGS
 The three pairs of legs called prothoracic leg,
mesothoracic leg and metathoracic leg are present.
 Each associated with a corresponding thoracic segment,
have a similar structure but differ in their relative size, i.e., they become
progressively larger from anterior to posterior.
 Male and female legs do not differ. But in prothoracic legs only
forelegs of males carry Sex combs, rows of thick dark bristles on the
first tarsal segment.
 Each leg is composed of five parts, from proximal to distal :
Coxa, trochanter, femur, tibia, and tarsus.
 Tarsus is composed of five tarsomeres.
ABDOMEN
 Both sexes have a pattern of dark transverse stripes on
the dorsal side of each abdominal segment.

 The abdomen of female has 8 segments, with 7 being

clearly visible. In male the last 3 abdominal segments are
fused, so that only 5 segments are clearly visible.

 Females have an abdomen with a pointed tip while

the male abdomen is rounded; additionally, the male
abdomen tends to curl inwards.

 Male external genitalia (epandrium) are larger, more

complex, and darker than the female external genitalia
(genital plates and ovipositor).
 SEXUAL DIMORPHISM
FEMALE MALE
 Larger in size  Smaller in size
 Tip of the abdomen is elongated  Tip of abdomen is rounded
 Abdomen has eight segments with 7 being clearly  Last three abdominal segments are fused , so only 5 segments
visible. are clearly visible.
 Abdominal tip is less darker than male  Abdominal tip is more darker than female
 Genital appears less darker  Male genital plate appears rougher and darker
 Sex comb is absent  Possess a sex comb on the 1st tarsal segment of 1st pair of
legs
 Testes in larvae & early pupae larger than ovaries in female
 Ovaries is smaller in larvae
FEMALE MALE
Lifecycle of Drosophila melanogaster
 Drosophila melanogaster have a short life cycle (2-3 weeks). For instance, at 20oC, the life cycle is completed in 14
or 15 days, but at 25oC, the cycle lasts about 10 days.
 They are holometabolous insects, life cycle is divided into 4 parts- embryo, larvae, pupa, adult.
 Egg produces larva, which eats and grows and at length becomes pupa and the pupa, in turn develops into an imago or
adult.
MATING-
 Female flies are unable to mate for several hours after becoming an adult
 If there is black spot or “meconium”, then the flies are considered as newly emerged and unmated flies because
meconium is the remains of their last meal before pupation.
 Drosophila males display a number of behaviours to achieve reproductive success. This includes:
Following the female
Tapping her with his forelegs
Contacting her genitalia with his mouthparts
Singing a species-specific courtship song using wings
Bending his abdomen to copulate
 Female flies agree by opening vaginal plate for copulation
 After successful copulation, mated females become temporarily sexually unreceptive to further copulatory attempts.
 EGG
Usually 0.5mm long
Penetration of spermatozoa into egg occurs through a small opening or micropyle, in the conical protrusion at the anterior
end, as the egg passes through the uterus.
Only one is fertilized.
Spermatozoa is stored by the female after mating.
After sperm enters, meiotic divisions are completed and the egg nucleus (female pronucleus) is formed.
The sperm nucleus and the egg nucleus then come into position side by side to produce the zygote nucleus, which divides
to form the first two cleavage nuclei.
Eggs are laid by the mother shortly after they are penetrated by the sperm.
A female adult lay 50-75 eggs per day for a total of approximately 400-500 eggs in 10 days.
 LARVAL STAGE
White, segmented, worm-shaped burrower with black mouth parts (jaw hooks) in the narrower head
region.
Pair of spiracles for tracheal breathing.
Young larvae must periodically shed skin in order to grow into adult size
After hatching from the egg undergoes two moults, so that the larval period consists of three instars.
During each period between moults, the larva is called an instar, i.e. the first instar is between
hatching and the first moult.

Both the size of the larva and the number of teeth on the dark coloured jaw hooks are an indication of
which instar the larva has reached.

After the second moult, the larva (now third instar) feed until ready to pupate.

At this stage, the larva crawls out of the food medium onto a relatively dry place, ceases moving, and
everts its anterior breathing spiracles.
Larvae adhere at dry place using the glue protein secreted by their salivary glands, following which they
undergo the last moult to begin the pupal stage
Pupa
 The most drastic changes in the transformation appears during pupal stage.
 The larva first leaves food and crawls onto the sides of the culture bottles for a
suitable place for pupation and comes to rest.
 The larva is now very sluggish, it everts it's anterior spiracles and become
motionless.
 Soon after everting its anterior spiracles, the larval body shortens and the cuticle
becomes hardened and pigmented.
 After 4 hours, a headless and wingless prepupa forms. This stage is followed by the
formation of the pupa with everted head, wing pads, and legs.
 Adult structures that have been present as small areas of dormant tissues since the
embryonic stage start to develop and show themselves.
 These tissues are called imaginal discs or anlagen and they are developed and reach
their full maturity during pupation.
 They go through histogenesis and form the extremities such as eyes, mouth parts,
antennae and genital apparatus, the body wall of imaginal head, thorax and abdomen.
Histolysis also occurs during pupal stage and salivary glands, the fat bodies, the
intestine and the muscles are destroyed.
 Adult
 Most of the structures of the adult fly develop during the larval period and take their final form during
eclosion, the act of emerging from the pupal case.
 The fruit fly is light in coloration, with expanded wings and an elongated abdomen.
 Within a few hours, the fruit fly darkens, extends its wings and expands its abdomen.
 Adults exhibit a typical insect anatomy, including compound eyes, three-part bodies (head, thorax, and
abdomen), wings, and six jointed legs. The various types of bristles and hairs found on the body.
 Approximately 48 hours after emerging from the puparia, female fruit flies are sexually mature and can
begin breeding and laying eggs.
 But the females wait for 10 to 12 hours
 Adult fruit flies are fertile for the entirety of their life spans.
 The removed flies are called virgins.
 Why are Drosophila such good model organisms?

 Many different characteristics of Drosophila melanogaster make it an ideal model organism. The major benefit of
using Drosophila is that there are also no ethical issues surrounding their use, which is common for mammal models
such as monkeys.

 1. Short lifespan
One of the main advantages is its short life cycle, allowing for a large quantity of flies to be produced within a short period.
Development of an adult fly only takes 10 days from fertilization. The female fly can produce up to 1500 eggs in its lifetime
thereby providing a constant supply of new Drosophila for genetic studies.

 2. Minimal culturing requirements

Another advantage of Drosophila is that they are very small and thus, very easy to maintain. Due to their small size and
minimal requirements, many Drosophila can be raised and tested within a small laboratory which does not have access
time, space or funding.
3.Genetic manipulation
Genetic factors also make this fly an ideal model organism. Drosophila melanogaster only has four pairs of
chromosomes compared to 23 pairs in humans. This simplicity was one of the reasons why they were first
used in genetic studies. Drosophila genes could be mapped easily to investigate genetic transmission.

4. Anatomical features
Drosophila has anatomical features [such as wings and eyes] which allow for easy characterization. These
genetic markers can be easily identified under a microscope.
Behaviors such as eating, mating, and sleeping that are observed in humans are also seen in Drosophila.
Therefore, the possible effect of genetic upon human behavior can also be assessed.
5.Genetic similarity with human genome
Many Drosophila genes are homologous to human genes and are studied to gain a better understanding of
what role these proteins have in human beings. Much research about the genetics of Drosophila over the last
50 years has resulted in a wealth of reference literature and knowledge about hundreds of its gene.
 HISTORY OF DROSOPHILA RESEARCH

 William E. Castle (Harvard University) used Drosophila in 1901, incorporating them into his work on the genetics of coat color in both mice
and guinea pigs. He started using Drosophila due to their small size and faster breeding rate.

 Frank E. Lutz (Cold Spring Harbor) selected wing venation changes, looking for interruptions, forking, or changes in the width of the veins.
As was characteristic of this Darwinian approach, for each generation all the offspring were tabulated for the varieties of subtle changes among
the total offspring.

 Thomas H. Morgan (Columbia University) inbred Drosophila for generations. His research led him to conclude that the white-eye
characteristic, which he named the white gene, must be carried on the X chromosome11 which, in turn, led to the discovery of sex-linked
inheritance.

 Hermann Joseph Muller won the Nobel Prize in 1946 in Physiology for the conclusion that ionizing radiation causes mutations such as
chromosomal rearrangements. Muller proved that treating the sperm of Drosophila with X-rays produced mutations in genes that were stably
inherited and could be followed through three or four further generations.

 Seymour Benzer first formed the basis of our understanding of circadian rhythms in Drosophila. In this assay, he quantitatively measured the
behavioral responses of flies to a light stimulus.


 TIMELINE OF DROSOPHILA
RESEARCH
 Genetics of Drosophila
 Drosophila melanogaster has four sets of chromosomes. the X and Y sex chromosomes, three pairs of autosomal
chromosomes 2 and 3, and 4. Chromosome 4 is very small, dot-like, and is known as the Dot chromosome.

 Female flies carry two X chromosomes and males carry a single X and Y chromosome. Females and males carry
two sets of autosomal second, third, and fourth chromosomes.

 The X chromosome, Y chromosome and Chromosome 4 are acrocentric whereas chromosomes 2 and 3 are
metacentric

GENOMICS
 Drosophila melanogaster genome sequencing was completed in March 2000.
 The Drosophila melanogaster genome sequencing project was led by the Berkeley Drosophila Genome Project
(BDGP) and the Whole Genome Shotgun effort was carried out in collaboration with Celera Genomics.
 The Drosophila genome is about 180 Mb in size and codes for approximately 13,600 genes
 SEX DETERMINATION
X Chromosome Autosome set X: A ratio Sex
 The Y chromosome is not necessary for the viability of
3 2 1.50 Metafemale
Drosophila; however, XO males lacking the Y chromosome
are sterile. In flies, the Y chromosome is not involved in 4 3 1.33 Metafemale
determining sex. Rather, it contains genes active in forming
sperm in adults. 4 4 1.00 Normal

 Furthermore, XXY flies are female, indicating that the Y 3 3 1.00 Normal female
chromosome plays no role in sex determination in
2 2 1.00 Normal female
Drosophila.
 Sex is determined by the ratio between the X chromosome 2 3 0.66 Intersex

and autosome in Drosophila.

1 2 0.50 Normal male

1 3 0.33 Metamale
 Gynandromorphs
 Gynandromorphs are individual animals that have both genetically male
and female tissues and often have observable male and female
characteristics. They may be bilateral, appearing to divide down the middle
into male and female sides, or they may be mosaic, with
patches characteristic of one sex appearing in a body part characteristic of
the other sex.
 In Drosophila, this can happen when an X chromosome is lost from one
embryonic nucleus. The cells descended from that cell, instead of being XX
(female), are XO (male). Because there are no sex hormones in insects to
modulate such events, each cell makes its own sexual “decision.” The XO
cells display male characteristics, whereas the XX cells display female
traits. This situation provides a beautiful example of the association
between insect X chromosomes and sex.
 Applications of Drosophila in different fields
 D. melanogaster acts as a model organism for biological research in studies of genetics physiology, developmental biology
(embryology), microbial pathogenesis, evolution, etc.

1. Genetical Studies
 D. melanogaster was among the first organisms used for genetic analysis.
 Presently, it is one of the most widely used and genetically best-known of all the eukaryotic organisms.
 Many basic principles of heredity such as sex linked inheritance, epistasis, multiple allelism and gene mapping are based on
studies in Drosophila.

2. Pathological studies
 About 75% of known human disease genes have a recognizable match in the genome of D. melanogaster. About 50% of fly
protein sequences have mammalian homologs. So D. melanogaster is being used as genetic model for several human diseases
such as Parkinson disease, Huntington's disease and Alzheimer's disease.
 The fly is also being used to study mechanisms underlying aging and oxidative stress diabetes and cancer, as well as drug
abuse.
3.Developmental Studies
 Embryogenesis in Drosophila is peculiar that the cleavage occurs in the syncitium so the early embryo contains
approximately 5000 to 6000 nuclei in an unseparated cytoplasm of the embryo.
 During gastrulation and early development, embryo undergoes well characterized morphogenetic movements including
germ band extension, formation of endoderm (gut), etc. due to which they are used in developmental studies.

4. Immunological studies
 Humoral immune response in Drosophila is a systemic response.
 It is mediated through the Toll and imd pathways which help in detecting the microbes.
 The Toll pathway in Drosophila has homologues of Toll- like pathways in mammals. Due to these similarities they are used
in immunological studies.

5.Behavioural studies
 Ron Konopka and Seymour Benzer (1971) described mutation- caused behaviour pattern changes in Drosophila. They
reported that the mutants have faster or slower or broken rhythms. They proposed that these mutations affect a group of
genes and their products which comprise biological clock.

 In recent years, Drosophila have been widely used in cancer studies and drug discovery process.
CURRENT RESEARCHES ONGOING ON
DROSOPHILA IN UNIVERSITY OF DELHI
 Surajit Sarkar – Neurobiology, Developmental genetics
 Mallikarjun Shakarad – Experimental evolution; life history trade offs, developmental rates and aging.
 R.Rajagopal –Gut microbe diversity and their functional role in insects.
 Dau Dayal Aggarwal – Genetics and population genetics
 Namita Aggarwal – Neurodegenerative diseases like Huntington’s disease

 This is not an exhaustive list

 Drosophila Culturing
 Drosophila melanogaster has proved to be an easily cultured and robust laboratory animal.
 Basic fly handling equipment includes a binocular microscope with a good light source, an etherizer or a CO2
plate for anesthetization, a fly pusher, an aspirator, a pounding pad, and a morgue. For most purposes flies can be
kept at room temperature , but one or two constant temperature rooms, preferably humidified, or incubators are
generally useful and are necessary for some techniques.
 Adult flies as well as larvae feed on fruit juices and yeast constitutes an important part of their diet .Therefore
Drosophila may be raised on any fermenting medium.
 The different types of medium routinely used for breeding Drosophila include cornmeal medium, banana jaggery
medium, sucrose dextrose medium and maltose corn medium. The composition of the food predominantly includes
sugar, yeast extract, dextrose and corn flour. Nipagin is also added to prevent bacteria and mold from growing on
food.
 They can be bred in glass bottles to obtain large numbers of the progeny.
A typical bench set up for work with Drosophila: (a)
stereomicroscope, (b) CO2 regulator, (c) chamber for
anesthesia, (d) paint brush, (e) porous pad connected to A bottle containing Drosophila larvae and
CO2 , (f) a fly morgue: a bottle containing methanol, and pupae
(g) a cold light source.
 Limitations of using Drosophila in genetics

 Despite the advantages of using Drosophila to investigate human diseases, there are
also several disadvantages. Firstly, the anatomy of the brain and other major organs
within the fly are very different from humans.
 Other issues include lack of methods to measures behavioral tendencies, lack of in-
depth cognitive abilities, lack of an adaptive immune system and dramatically
different drug effects when compared to human studies.
REFERENCES

 https://doi.org/10.1016/j.cub.2018.11.034
 Atlas of Drosophila Morphology ( University of Cambridge) .
 Wonder animal model for Genetics studies : Drosophila M. – it’s life cycle and breeding method : A review.
 Experiments with Drosophila for Biology course ( Indian Academy of science Benglore)
 University of Kentucky: Drosophila ( fruit fly )
 https://doi.org/10.1007/978-981-13-0529-0_1
 Doi: 10.1146/annurev.genom.4.070802.110323
 Developmental Biology by Scott F. Gilbert
 https://www.news-medical.net/life-sciences/Drosophila-as-a-Model-Organism.aspx
 https://doi.org/10.1016/S1369-7021(11)70113-4
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