Metamorphosis

Metamorphosis
post embryonic extension of the
developmental potential
involves dramatic changes in habit,
habitat, morphology, physiology and
behaviour of youngone that is
transformed into the adult
 INTRODUCTION
• Direct development: Hatched organism (juvenile)
resembles an adult, except that it is small in size.
Only growth and sexual development occur after
hatching
• Viviparity: Hatched organism grows and develops
inside the body of the mother. Remaining growth
takes place after birth.
• Indirect development: Egg develops into a
strikingly different organism (larva) from the adult.
Metamorphosis occurs in at least 17 phyla of the animal kingdom,
including Porifera, Cnidaria, Platyhelminthes, Mollusca, Annelida,
Arthropoda, Echinodermata, and Chordata.
 TYPES OF METAMORPHOSIS

• Retrogressive metamorphosis: Change of more

complex organization into simple organization
(Ascidians)- Adults: sessile, no locomotary
organelles.

• Progressive metamorphosis: Change of simple

organization into more complex organization
(Amphibians and Insects)
RETRAGRESSIVE METAMORPHOSIS
(TUNICATES)
PROGRESSIVE METAMORPHOSIS (AMPHIBIANS)
PROGRESSIVE METAMORPHOSIS (INSECTS)
Process of metamorphosis implies several
universal events:
• Destruction of specific larval structures
(abdominal legs of caterpillars, gills and tail of
tadpoles)
• Adaptive remodelling of tissues that persist in
the adult stage (nervous system, excretory
organs)
• Development OR Construction of structures
unique to adults (wings in insects, lungs in
amphibians)
• Most spectacular metamorphosis seen
occurs in insects and amphibians under
precise hormonal control.
Metamorphosis
in Insects
 INSECT METAMORPHOSIS
• Special form of moulting
• Moulting-casting off an old cuticle and the
acquisition of a new one
• Change of cuticle is necessary for growth and
metamorphosis to occur, because the fully
sclerotinized cuticle is rigid and does not
expand
 INSECT METAMORPHOSIS
• Molting period is punctuated by two events, apolysis and
ecdysis, that define insect development.

• Apolysis: Separation of the epidermal cells from the

cuticle, marks the beginning of the molt and the next
instar.
• Ecdysis: marks the beginning of the next stadium.

• At the apolysis following the second instar, the insect

enters the third instar but is still in the second stadium
until after ecdysis.
 INSECT METAMORPHOSIS

• Instar is defined as the period between two

apolyses and begins when the insect first
becomes detached from its old skin
• Instar that is hidden under the old, unshed cuticle
before ecdysis is referred to as the pharate
instar.
• Stadium represents the interval between one
ecdysis and the next.
Insects are hard-bodied, they cannot grow larger
gradually. Instead they grow larger in steps by shedding
the hard exoskeleton for a brief period of expansion. The
brief periods between or within stages are called molts.
Insects are soft-bodied and vulnerable during this time.
Cicada Molting Exoskeleton
 AMETABOLOUS DEVELOPMENT
• Direct Development
• No larval stage
• Pronymph stage immediately after hatching
• Young insects look like small adults but genitalia are
absent.
• After each molt, they are bigger, but unchanged in
form.
• Both the immatures and adults occupy the same habitats.
 Without Metamorphosis
adult
nymphs
egg

Without meta

First type is "without" metamorphosis (ametamorphosis or

ametabolic). The young resemble adults except for size. Eg.
Silverfish (Thysanura) and Springtails (Collembola).
 HEMIMETABOLOUS
DEVELOPMENT
• Also called incomplete/gradual metamorphosis
• Immatures lack wings and genitalia and the extent
of their metamorphosis is the development of
these structures in adults.
• Both have compound eyes, but those in adults
may have more optical units, or ommatidia.
• Also show differences in cuticular structure.
• Nymphs eat the same food that the adult insect
eats
 Gradual Metamorphosis
Second type is "gradual" metamorphosis seen in grasshoppers
(Orthoptera), termites (Isoptera), thrips (Thysanoptera), and true bugs
(Hemiptera). Life cycle starts as an egg, but each growth, or nymphal
stage looks similar, except it lacks wings and the reproductive capacity
that the adult possesses.
Gradual meta

egg nymphs adult

Pronymph Nymph
Instar 1 Nymph Instar
2

Adult
Hemimetabolous Development
in Cockroach
Life cycle of a hemimetabolous insect, showing the eggs,
nymphs of the five instars, and the adult bug
 Complete Metamorphosis
Third type is "complete" metamorphosis. Life cycle has the four
stages of egg, larva, pupa, and adult. Each stage is quite distinct.
Found in butterflies (Lepidoptera), beetles (Coleoptera), flies
(Diptera), and bees, wasps, and ants (Hymenoptera).

egg larvae pupa adult

 Growth of Larval Instars of
Silkworm

First
Instar
Fifth
Instar

Third 5000-fold
Instar
increase!
Larva (Instar 1) Larva (Instar 2)

Pupal molt

Imaginal molt
Pupa
Adult
Heteromorphosis or hypermetamorphosis:
Successive larval instars occupy different habitats
and display a marked anatomical variation.
This is a deviation from more conventional
holometaboly with two or more larval forms with
active first-instar larvae and more inactive grub like
late instar larvae
Some Coleoptera, Hymenoptera, Neuroptera, and
all Strepsiptera,.
 IMAGINAL DISCS
• In holometabolous insects, the transformation
from juvenile into adult occurs within the pupal
cuticle.
• Most of the old body of the larva is systematically
destroyed by APOPTOSIS
• New adult organs develop from undifferentiated
nests of cells called the IMAGINAL DISCS.
 IMAGINAL DISCS
Within any larva, there are two distinct
populations of cells:
• i) The larval cells, which are used for the
functions of the juvenile insect, and
• ii) The thousands of imaginal cells,
which lie within the larva in clusters,
awaiting the signal to differentiate.
 Types of Imaginal Cells

Clusters of
Imaginal Histoblast
Imaginal
Discs Nests
cells
Histoblast Nests
• Histoblast nests are groups of imaginal
cells that will form the adult abdomen.

Clusters of Imaginal Cells

• Within each organ they will proliferate to
form the adult organ as the larval organ
degenerates.
 Imaginal Discs

• Cells of the imaginal discs will form the

cuticular structures of the adult, including
the wings, legs, antennae, head, thorax
and genitalia.

• Imaginal discs can be seen in the newly

hatched larva as local thickenings of the
epidermis.
Imaginal Discs in Drosophila
Control of Insect
Metamorphosis
 Wigglesworth’s Experiments
Tiny first instar
First instar Fused to develops cuticle,
decapitated molting 5th body structure and
(Head removed) instar genitalia of adult

Conclusion: Blood-borne hormones are responsible

for the induction of metamorphosis
Corpora At next molt, 3rd
allata instar turns into
removed precocious
from 3rd adult
instar
Corpora Corpora 5th instar molts
allata allata into extremely
removed transplanted large “6th instar”
from 4th into 5th instead of adult
instar instar

Conclusion: Corpora allata, near the insect brain,

produces a hormone that counteracts tendency to
undergo metamorphosis (JUVENILE HORMONE)
Normal 5th Normal “6th
instar adult instar”
 Demonstrated
involvement of
hormone from
anterior body &
timing of
release.

Confirmed
location of
hormone in
brain.

Kopec’s simple ligation & debraining experiments (ca. 1919).

 HORMONES OF INSECT
METAMORPHOSIS
• Sequence of events in metamorphosis is under
precise hormonal control (Kopec).
• Five major insect hormones:
• Prothoracicotrophic hormone (PTTH)
• Juvenile hormone (JH) or Neotinin
• Ecdysone or Ecdysterone or -ecdysone (active
form) or 20-hydroxy ecdysone (20E) (Butenandt
& Karlson 1954)
• Eclosion hormone
• Bursicon
 PTTH Juvenile Ecdysone Eclosion Bursicon
Hormone

Secreted Brain Corpora Prothor- Corpora Brain,

by allata acic cardiaca nerve
gland cord
Chemical Small Fatty Chole- Peptide Protein
nature protein acid sterol
deriv.
Acts on Retention Induces Shedding Harden-
Primary prothorac- of larval Molting of cuticle ing of
Role ic gland to characters new
secrete cuticle
ecdysone
 ENDOCRINE CONTROL OF
MOLTING
• Basic hormonal mechanism is identical
whether the molt is larval-larval, larval-adult,
larval-pupal, or pupal-adult.
• What does differ during metamorphosis is the
nature of the cuticle that the insect molts to
and that is regulated by the presence or
absence of juvenile hormone.
• In the presence of a high concentration
of JH, the epidermal cells secrete a
larval cuticle.
• In the presence of a low concentration
of JH, the epidermal cells secrete a
pupal cuticle.
• When JH concentration is zero or
negligible, the epidermal cells secrete
the adult or imaginal cuticle.
 Concentration of JH

Larva
Pupa
Adult
 Functions of Juvenile Hormone In
the Adult
The concentration of JH rises once the adult insect
emerges.
In the adult, JH serves the following functions:

• In the males of some insect species, juvenile

hormone promotes development of accessory
sexual organs.

• In many female insects, it induces yolk synthesis

and promotes maturation of the egg.
 ECDYSONE
• There are two major pulses of 20E during
metamorphosis.
• The first pulse occurs in the third instar larva and
triggers the initiation of “prepupal” morphogenesis
of the leg and wing imaginal discs as well as the
death of the larval hindgut. The larva stops eating
and migrates to find a site to begin pupation.
• The second 20E pulse occurs from 10-12 hours
later and tell the prepupa to become a pupa. The
head inverts and salivary glands degenerate.
 Second
pulse
First
pulse

The Concentration of Ecdysone during

Metamorphosis of Drosophila
 Eclosion hormone and bursicon
are responsible for promoting the
terminal phase of molting process

• Shedding of the • Bursicon acts

cuticle of the to harden and
pupa is triggered
sclerotize the
by the eclosion
hormone. new cuticle
(b) Flow diagram of the events
initiating molting in an insect.
(c) Flow diagram of the events
of insect metamorphosis as
regulated by a decrease in the
production of juvenile hormone.
Endocrine control of
molting in insects having
complete metamorphosis.
Juvenile hormone and
ecdysone interact to
control molting and
pupation.
Many genes are activated
during metamorphosis, as
seen by puffing of
chromosomes.
Puffs form in sequence
during successive molts.
Changes in cuticle thickness
and surface characteristics
are shown at right.
Changing activities of the epidermis during the fourth and fifth larval instars and prepupal
(pharate pupal) development in relation to the hormonal environment. The dots in the epidermal
cells represent granules of the blue pigment insecticyanin. ETH (ecdysis triggering hormone);
EH (eclosion hormone); JH (juvenile hormone); EPI, EXO, ENDO, deposition of pupal epicuticle,
exocuticle, and endocuticle, respectively. The numbers on the x-axis represent days.
Metamorphosis
in Amphibians
Extent of Metamorphosis in Amphibians

• Anurans • Urodeles
• Frogs • Salamanders
• Toads • Newts
Undergo drastic Metamorphosis is
metamorphosis. not as drastic.
Tail is not lost.
Tail is resorbed.
Limbs poorly
Limbs well
developed; are
developed; are efficient
efficient jumpers swimmers.
on land.
In amphibians (frogs), metamorphosis is
generally associated with the changes that
prepare an aquatic organism for a primarily
terrestrial existence

A gill-breathing tadpole larva subsisting on a

herbivorous diet is transformed in to lung-
breathing, carnivorous adult.
 Tadpole with
external gills
Eggs

Tadpole with
internal gills
Adult Frog

Tadpole with
hind limb buds
Froglet

Tadpole with
Tadpole with all legs
hind legs
TADPOLE LARVA CHARACTERS

• Herbivorous
• Mouth has horny beaks with rows
of horny teeth
• On each side of head a fold of skin,
the operculum, which covers the gill
• Epidermis with large pigment cells
• Intestine is long and coiled
• Ammonotelic
• RBC form from the kidney
• Visual pigment in retina is
porphyropsin
 Metamorphic Changes

Adaptive
Destructive Constructive
Remodelling
 Destructive or Regressive Changes

• Tail resorption
• Internal gills, gill clefts and peribranchial cavities
disappear.
• Opercular fold falls off.
• Horny teeth, ventral suckers, labial fringes and horny lining
of jaws are shed.
• Lateral line sensory system disappears.
• Some blood vessels disappear.
• Apoptosis inducing enzyme, CASPASE-9 is important in
causing cell death of different organs
 Destructive or Regressive Changes
• Autolysis
• Reduction of the organs by the action of
amoeboid macrophages
• Many epithelial cells are dependent on their
attachment to the ECM to prevent apoptosis.
The rapid apoptosis that occurs with the loss
of ECM attachement is called as ANOIKIS
• Degrowth
• Reduction in the body mass
CONSTRUCTIVE OR PROGRESSIVE CHANGES

New structures that were absent in the larva develop in the adult.
• Limb development
• Tongue is developed from the floor of the mouth
• Bony skull
• Lungs
• 3-chambered heart
• Middle ear and tympanic membrane
• Eyelids and nictitating membranes in the eye.
 ADAPTIVE CHANGES
Some organs function in both the tadpole and the adult, but
undergo some degree of alteration and differentiation to
better suit the needs of the adult.
• Hardening, keratinization and pigmentation of the skin.
Multicellular mucous and serous glands develop.
• Widening of the mouth, re-shaping of the snout, and re-
positioning of the eyes (lateral position to front of
head).
• Trunk becomes narrower than the head.
 ADAPTIVE CHANGES
• Intestine is reduced from 9 times the body length to twice
the body length (carnivorous mode of life). Guided by the
enzyme called metaloproteinase stromelysin-3
• Tongue becomes more muscular.
• Gill arches become modified into the hyoid apparatus
• Brain becomes highly differentiated.
• Cartilage skull changes to bony skull
• Middle ear and tymphanic membrane develops
Intestine before and after
metamorphosis
contralateral contralateral & ipsilateral
 BIOCHEMICAL CHANGES
• Porphyropsin (retinene2) to rhodopsin (retinene1)
• Tadpole haemoglobin (HbF) to adult haemoglobin (HbA) (binds
oxygen more slowly and releases it more rapidly)
• Erythropoiesis shifts from liver or kidney to bone marrow
• Peptic activity starts in the stomach for the digestion of animal
tissue.
• Endocrine function of pancreas starts. It begins to secrete insulin
and glucagon
• Reorganization of the liver for production of appropriate enzymes
of the ornithine urea cycle (synthesize enzymes necessary to
create urea from CO2 and NH3)
• Synthesis of melanin and serotonin in skin
 STAGES OF METAMORPHIC EVENTS
• Grant (1978)
• Premetamorphic period- tadpole growth with little
metamorphosis (little hind limb growth takes place)
• Prometamorphic period: Body grows at a reduced
rate. The ratio of hind limb length to body length
increases rapidly
• Metamorphic climax: Cessation of growth
accompanied by rapid changes in limbs, tail, head
and internal organs.
Premetamorphic Tadpole

Prometamorphic Tadpole
(growth of hindlimbs)

Onset of the Metamorphic Climax

(Eruption of forelimbs, retraction
of tail fin)

Climax stages
 HORMONAL CONTROL OF
AMPHIBIAN METAMORPHOSIS
Guder-natsch (1912) - by thyroid hormones

Experiment: If tadpoles are fed on diets of various tissues,

ovary, liver, thymus, brain, pancreas, spleen, pituitary, and
thyroid — those fed on thyroid gland develop limbs and
lose their tails long before the others.

Conclusion: A secretion of the thyroid gland is responsible for

triggering metamorphosis.
Removal or destroying the thyroid rudiment from
the early tadpoles (thyroidectomy), the larvae
never metamorphosed, instead they become
giant tadpoles. Allen (1916)
Hormones Involved in Amphibian Metamorphosis

Thyroxine (T4) Tri-iodothyroxine (T3)

 SEQUENCE OF EVENTS
1. Thyroid gland secretes a large complex protein-
thyroglobulin, which is made of T4 (Thyroxine)
2. T4 is released into the blood from the thyroid gland
3. Uptake of T4 by target tissue
4. Conversion of T4 into active hormone T3 by the
target tissue
5. Binding of T3 to receptors in the nucleus (Thyroid
hormone receptors)
6. Changes in gene expression
7. Degradation of T3 in the target tissue
 Conversion of T4 to T3

• Occurs in the cells of the target tissue

• Catalyzed by enzyme: Type II deiodinase
which removes 1 iodine atom from outer
ring of T4 to convert it to T3.
 I
e seI
y p
T i na
i o d
De

T4 T3
 Action of T3

•T3 exerts its effects by binding to thyroid hormone

receptors (TRs).
•TRs are members of the superfamily of steroid
hormone receptors.
•This family of receptors function as transcription
factors by regulating the messenger RNA levels of
specific genes.
 Pre- Pro- Climax
metamorphosis metamorphosis
 Degradation of T3

• Occurs within the cell of the target tissue

• Catalyzed by enzyme Type III deiodinase
• The enzyme removes 1 iodine atom from the
inner ring of T3 and converting it into an inactive
compound
• Eventually metabolized to amino acid tyrosine.
 Role of Prolactin
• Hormone prolactin inhibit the up-regulation of TRα
and TRβ mRNAs.
• As a result the tail is not resorbed, and the adult-
specific keratin gene is not activated .
• Injections of prolactin stimulate larval growth and
inhibit metamorphosis.
Thyroid hormones triiodothyronine (T3) and thyroxine (T4) regulate the metamorphosis of an aquatic
frog tadpole into a semiterrestrial or terrestrial adult. The anterior pituitary secretes thyroid-
stimulating hormone, which regulates thyroid gland activity. During the premetamorphosis (tadpole)
stage, the pituitary and thyroid glands are relatively inactive. This keeps the concentration of thyroid-
stimulating hormones, T3 and T4, at low concentrations. The high prolactin concentration in tadpoles
stimulates larval growth and prevents metamorphosis. During metamorphosis, the concentrations of the
thyroid hormones markedly increase, and prolactin decreases. These hormonal fluctuations induce rapid
differentiation, climaxing in the adult frog.
 TISSUE REACTIVITY
• Various organs of the body respond differently to
a single thyroxin agent
• Same stimulus will cause certain tissue to
degenerate and others to grow and differentiate
• Such diverse reaction of tissues is due to
• Competence of tissues
• Threshold value of different larval tissues
 Threshold concept
• Kollros (1961)
• Different larval tissues have different threshold
value (different concentrations) for thyroxine.
 THRESHOLD CONCEPT
Shortening of
Dilute intestines
solution of
thyroid Accelerated
hormones hind limb
growth

High Tail regression

concentration of before hind limbs
thyroid hormones are formed.
 HETEROCHRONY
• Changes, over evolutionary time, in the rate or timing of
developmental events.
• Two types-
(a) Allometry- Different organs of the body growing at
different rates
(b) Pedomorphosis or Juvenification- Change in which the
adults of a species retain traits previously seen only in
juveniles. A paedomorph is an organism that keeps
physical and mental characteristics of its childhood in its
adulthood
 Paedomorphosis
• Two types
• Neoteny- Somatic cells develop slower
whereas germ cells develop in normal time.
Axolotl larva of Ambystoma (tiger salamander)
• This larva does not undergo metamorphosis in
nature because its pituitary gland does not
release the TSH
 Paedomorphosis
• Progenesis- Somatic cells develop normal
time whereas germ cells develop in faster time.
Eg. Bolitoglossa occidentalis (tropical
salamander)
• Direct development: Some animals
accelerate their development by abandoning
their larval forms. Eg. Eleutherodactylus coqui
 Paedomorphosis
• Two types
• Obligatory- Permanent larvae. The developing
tissue fails to respond to the TH. Eg. Necturus
• Facultative- Temporary larvae. Larvae remains
aquatic, gill breathing, and fully reproductive,
unless the water begins to dry up. Eg. Axolotl
larva of Ambystoma (tiger salamander)
 Neoteny

White Axolotl Necturus larva

Progenesis

Bolitoglossa occidentalis