Instinctive Behaviour

•Instinct is the inherent disposition of a Living

organism toward a particular behavior. Instincts are
unlearned, inherited fixed action patterns of responses
or reactions to certain kinds of stimuli.
•Examples of instinctual fixed action patterns can be
observed in the behavior of animals, which perform
various activities (sometimes complex) that are not
based upon prior experience and do not depend on
emotion or learning, such as reproduction, and
feeding among insects.
•Other examples include animal fighting, animal
courtship behavior, internal escape functions, and
building of nests.
• Instinctual actions - in contrast to actions based
on learning which is served by memory and
which provides individually stored successful
reactions built upon experience - have no
learning curve, they are hard-wired and ready to
use without learning, but do depend on
maturational processes to appear.
• Biological predispositions are innate biologically
vectored behaviors that can be easily learned.
For example in one hour a baby colt can learn to
stand, walk, and run with the herd of horses.
• Learning is required to fine tune the neurological
wiring reflex like behavior.
• True reflexes can be distinguished from instincts
by their seat in the nervous system; reflexes are
controlled by spinal or other peripheral ganglion,
but instincts are the province of the brain.
• Technically speaking, any event that initiates an
instinctive behavior is termed a key stimulus (KS)
or a releasing stimulus. Key stimuli in turn lead to
innate releasing mechanisms (IRM), which in
turn produce fixed action patterns (FAP).
• More than one key stimulus may be needed to
trigger an FAP. Sensory receptor cells are critical
in determining the type of FAP which is initiated.
• For instance, the reception of pheromones
through nasal sensory receptor cells may trigger
a sexual response, while the reception of a
"frightening sound" through auditory sensory
receptor cells may trigger a fight or flight
response.
• Instinctive behavior can be demonstrated
across much of the broad spectrum of animal
life. According to Darwin's theory of evolution
by natural selection, a favorable trait, such as
an instinct, will be selected for through
competition and improved survival rate of life
forms possessing the instinct. Thus, for
evolutionary biology, instincts can be
explained in terms of behaviors that favor
survival.
• Any repeated behavior can be called "instinctual." As
can any behavior for which there is a strong innate
component. However, to distinguish behavior beyond
the control of the organism from behavior that has a
repetitive component
• innate behavior = behavior determined by the "hard-
wiring" of the nervous system. It is usually inflexible, a
given stimulus triggering a given response. A
salamander raised away from water until long after its
siblings begin swimming successfully will swim every
bit as well as they the very first time it is placed in the
water. Clearly this rather elaborate response is "built
in" in the species and not something that must be
acquired by practice.
• A number of criteria were established which
distinguishes instinctual from other kinds of
behavior. To be considered instinctual a behavior
must a) be automatic,
• b) be irresistible,
• c) occur at some point in development,
• d) be triggered by some event in the environment,
• e) occur in every member of the species,
• f) be unmodifiable, and
• g) govern behavior for which the organism needs no
training (although the organism may profit from
experience and to that degree the behavior is
modifiable).
• The absence of one or more of these criteria
indicates that the behavior is not fully instinctual.
• Instincts do exist in insects and animals as can
be seen in behaviors that can not be changed by
learning. Psychologists do recognize that humans
do have biological predispositions or behaviors
that are easy to learn due to biological wiring, for
example walking and talking.
 Fixed Action Patterns
• FAP are stereotypic and inborn behavior. They
are genetically determined and present a
complex interaction with environmental stimuli.
The Dutch ethologist Niko Tinbergen, Nobel
Prize, was one of the first to study FAP in
vertebrates, such as seagulls, ducks and so on.
• A classical example of FAP studied by Tinbergen
is the goose behavior of picking eggs up.
• When the female notices an egg outside the nest
(key stimulus) , it begins a repeated movement
to drag the egg with its beak and neck.
• However if the egg slides off or if it is removed by
the researcher, the goose continues to repeat
the stereotypic movements even if the egg is
absent, until it reaches the nest, when then it
does it all over again.
• Therefore, FAP seems to correspond to a fixed
neural circuitry elicited by the overall trigger
stimuli.
• Female goose behavior of picking eggs up. When it sees an egg outside
the nest (key stimulus), it begins a repeated movement of dragging the
egg with its beak and neck. However, if the eggs slides off or if it is
removed by the researcher, the goose continues to repeat the
stereotypic movements even if the egg is absent, until it reaches the
nest, when then it does it all over again. FAP seems to correspond to a
fixed neural circuitry elicited by the overall trigger stimuli
 Releasers of Instinctive Behavior
• So once the body is prepared for certain types of
instinctive behavior, an external stimulus may be
needed to initiate the response. N. Tinbergen
(who shared the 1973 Nobel Prize with
Konrad Lorenz and Karl von Frisch) showed that
the stimulus need not necessarily be appropriate
to be effective.
• During the breeding season, the female three-
spined stickleback normally follows the red-
bellied male (a in the figure) to the nest that he
has prepared.
• He guides her into the nest (b) and then
• prods the base of her tail (c).
• She then lays eggs in the nest.
• After doing so, the male drives her from the
nest, enters it himself, and fertilizes the eggs
(d).
• Although this is the normal pattern, the
female will follow almost any small red object
to the nest, and
• once within the nest, neither the male nor any
other red object need be present.
• Any object touching her near the base of her
tail will cause her to release her eggs.
• It is as though she were primed internally for
each item of behavior and needed only one
specific signal to release the behavior pattern.
• For this reason, signals that trigger instinctive
acts are called releasers.
• Once a particular response is released, it usually
runs to completion even though the stimulus has
been removed.
• One or two prods at the base of her tail will
release the entire sequence of muscular actions
involved in liberating her eggs.
• Chemical signals (e.g., pheromones) serve as
important releasers for the social insects: ants,
bees, and termites. Many of these animals emit
several different pheromones which elicit, for
example, alarm behavior, mating behavior, and
foraging behavior in other members of their
species.
• The mammary glands of domestic rabbit mothers
emit a pheromone that releases immediate
nursing behavior by their babies (pups). A good
thing, too, as mothers devote only 5–7 minutes a
day to feeding their pups so they had better be
quick about it.
Male robin Female robin
• The studies of Tinbergen and others have shown
that animals can often be induced to respond to
inappropriate releasers.
• For example, a male robin defending its territory
will repeatedly attack a simple clump of red
feathers instead of a stuffed robin that lacks the
red breast of the males.
 Female flicker

Male Flicker
• The male common flicker Colaptes auratus a
type of woodpecker found throughout North
America has a black stripe or moustache
extending from its bill along the sides of its
head.
• It is presence of this moustache on a male
entering another’s territory that is specific
releaser for aggressive behaviour.
• If such moustache is painted on a female of a
mated pair male actually attacks his mate. If
moustache is removed his aggression ceases.
• Although such behavior seems inappropriate to
our eyes, it reveals a crucial feature of all animal
behavior: animals respond selectively to certain
aspects of the total sensory input they receive.
• Animals spend their lives bombarded by myriad
sights, sounds, odors, etc.
• But their nervous system filters this mass of
sensory data, and they respond only to those
aspects that the evolutionary history of the
species has proved to be significant for survival.
• Ethologists, those who study animal behavior,
believe that every species have routine
movements that appear to be automatic in a way
that relates to their structural systems
• . Konrad Lorenz, one of the leading scholars in
this field, names these patterns as "Fixed Action
Patterns”.
• Further defining instinctive behavior, ethologists
found particular characteristics, which include
inherent structured systems and the adaptive
functions.
• Inherent structured systems are highly correlated
with innate activity; many behaviors of animals are
sufficiently unvarying and provide as particular
characteristics of bodily structures.
• For example, the web spinning movements of
spiders are a direct usage of its bodily construction.
Or, the burrowing habits of marine worms employ
operations of structure.
• Such movements that are typical to instinctive
behavior include, eating, care of body surface,
escape from predators, social behavior, and sexual
interaction.
• Most of these innate activities involve the particular
usage of a physical structure that is specific to each
species.
• Not just simple responses to an external stimulus
play a role in instinctive behavior; instinctive activity
involves sequences of behavior that run a
predictable course. These behaviors may last
seconds, minutes, hours or even days.
• Exemplifying this, we can refer to a particular
species of digger wasp, which finds and captures
only honeybees
• With no previous experience, a female wasp will
unearth an intricate burrow, find a bee, paralyze it
with a careful and precise sting to the neck, pilot
back to her discreet home, and, when the latter has
been supplied with the correct number of bees, lay
an egg on one of them and seal the chamber.
• The female wasp's whole behavior is designed so
that she can function in a single specialized way.
• Ethnologists believe that this entire behavioral
sequence has been programmed into the wasp
by its genes at birth thus resulting the high
correlated sequences between heredity and
instinctive behavior
• Given that instinctive behavior supposes to be
hereditarily based, and therefore shaped by the
forces of natural selection, it follows that most of
the outcomes of instinctive activity contribute to
the preservation of an individual or to the
continuity of the species; instinctive activity
tends to be adaptive, which implies the
alteration of a living organism to its surroundings
• There are two different types of adaptation;
one, which involves the accommodation of an
individual organism to a sudden change in
environment and the other, occurs during the
course of evolution and hence is called
evolutionary adaptation.
• Overall, one of the main distinctive features of
instinctive activity is the ability to react to an
external stimulus the correct way the first
chance (and every time thereafter) the animal
receives.
• This feature distinguishes this particular behavior
from what ethnologists call learned behavior,
which scientists have discovered are actions that
take place from conditioning an animal to learn
the right way.
 Genetic basis of egg laying
behaviour in Aplysia
• Aplysia is a gastropod mollusk of the order
Opisthobranchia, meaning “gills behind” (from
Greek opisthen for behind, plus branchia for
gills) and referring to the location of the gill of
Aplysia behind the heart. Aplysia is often called
the “sea hare” because the sensory tentacles
on the top of its head, called rhinophores, are
somewhat reminiscent of rabbit's ears. Unlike
other gastropods, Aplysia doesn't have a large
external shell in which to retreat.
• It has a small, flat, vestigial shell, the
consistency of cardboard, covering its viscera
and nominally protecting the heart and other
internal organs. The mantle, or main muscular
region on its back, is modified into 2 sections
that look a little like wings, called parapodia,
which cover the gill and help to channel water
over it so that the animal can breathe. Water
that has passed over the gills to extract
oxygen is directed out from between the
parapodial flaps via a tube, also modified from
the mantle, called the siphon.
• Nerve Central
• Aplysia californica are often studied by
neurobiologists because of the simplicity of
their nervous system. An animal's entire
nervous system contains about 20,000
neurons, making it easier to study than that of
higher animals like human beings, which can
have more than 100 billion neurons (in an
adult brain).
• Today we saw something strange on our sea
hare (we call it Shrek !). It is a strange long thing
(like a tentacle) on the head (front of the sea
hare). It does not seem to bother him, because
he still moves around. At the end of this tentacle
thing is a white piece, still attached to the
tentacle. To me it nearly looks like the fetus of
something (similar features). See attached
photo.
• We would appreciate it if you could explain to us
what that is !
• ANobel Cause
• Dr. Eric Kandel, a professor at Columbia
University, in 2000 was awarded with the Nobel
Prize in Physiology or Medicine for his work using
Aplysia californica to demonstrate fundamental
ways in which nerve cells alter their
responsiveness to chemical signals to produce a
coordinated change in behavior.
• Experiments can often determine if behaviors
have a genetic basis:
• Aplysia experiments show both nervous system
and endocrine system control body responses.
– a. Egg-laying behavior of Aplysia snail involves
sequence of movements.
– b. After copulation, snail extrudes long string of egg
cases.
– c. Takes egg case string in mouth, covers it with
mucus, waves head to wind string to attach it to
rocks, etc.
• The reproductive behaviour of sea hare Aplysia
involves a number of Fixed Action Patterns such
as the coordinated action involved in the egg
laying.
• When snail is about to lay eggs it stops feeding
and moving. Physiological changes involve are
heart and respiratory rates increase and the
muscles of the reproductive ducts contract as the
animal expels a string of eggs.
• The record holding sea hare laid a string 17,520
cm long at the rate of 41,000 eggs/minute
• When the egg string first appears, the snail grasps
it in a fold of its upper lip and, waving it’s head in
a stereotyped pattern, helps to pull the string out
and winds it into an irregular mass.
• During this process the egg string is coated with
sticky mucus. Finally the tangled egg string is
attached to a rock or other firm support with a
characteristic wave of the head.
• The neuro endocrine link between the genes and
egg-laying behaviour is studied by Dr. Eric Kandel
– d. Scientists isolated egg-laying hormone (ELH);
triggers egg-laying even without mating.
– e. ELH is 36-amino acid protein; diffuses into
circulatory system causing smooth muscle
contractions resulting in egg expulsion.
– f. Using recombinant DNA techniques, researchers
isolated ELH gene; gene product is protein with
271 amino acids that cleaves into 11 products
including ELH.
• The neuroendocrine link between the genes and
egg laying behaviour is known.
• Two small proteins, peptide A and peptide B are
produced by the atrial gland in the reproductive
system.
• When these peptides are injected in to an
animal, they stimulate two clusters of neurons
on top of the abdominal ganglion, called bag
cells, to release other peptides.
The bag cell neurons are located in two clusters of
~200-400 cells each at the rostral end of the
abdominal ganglion of Aplysia (Kupferman, 1967;
Kupfermann & Kandel, 1970; Pinsker & Dudek,
1977).
Their primary function is to act as a master switch
for a series of reproductive behaviors that
culminates in egg laying.
The electrophysiological state of these neurons
determines whether egg-laying behaviors will
occur in response to environmental stimuli.
• The series of reproductive behaviors is
triggered by the release of several
neuropeptides (Arch & Smock, 1977; Pinsker
& Dudek, 1977).
• Their major neurotransmitter is egg-laying
hormone (ELH), a 36-amino acid peptide that,
like many other neuropeptides, is amidated at
its C-terminus, conferring resistance to
extracellular proteases (Chiu et al., 1979;
Scheller et al., 1983). (Rothman et al., 1983a).
• ELH has actions on numerous other neurons in
the abdominal ganglion (Branton et al., 1978), as
well as on peripheral targets, particularly the
ovotestis
• ELH is synthesized as part of a larger precursor
(pro-ELH, Fig. 1), which also encodes several
smaller neuroactive peptides, that are termed -, -
and -BCP (bag cell peptides).
• Figure 1: Organization of pro-ELH, the precursor
to ELH and the small BCPs.
These smaller BCPs are also secreted locally to
alter the firing patterns of neurons in the
abdominal ganglion,
but they do not undergo any posttranslational
modification
and their actions are rapidly terminated by
proteases in the extracellular space
(Rothman et al., 1983b; Mayeri et al., 1985;
Pulst et al., 1986 & 1987; Sigvardt et al., 1986;
Kauer et al., 1987; Fisher et al., 1988;
Nagle et al., 1990).
• Several other pepdides, including -BCP are also
secreted during afterdischarge (Hatcher &
Sweedler, 2008),
but their functions are not yet understood.
• Interestingly, different combinations of
neuropeptides are packaged into different
secretory granules and are targeted to different
branches of the bag cell neurons (Sossin et al.,
1990; Li et al., 1998),
• presumably in order to coordinate the multiple
actions of these neurons on both peripheral and
central targets.
• Secretion of ELH and the BCPs from the bag cell
neurons normally occurs in response to a
prolonged burst of action potentials, termed an
afterdischarge. In the absence of stimulation, the
cells maintain negative resting potentials and
display no spontaneous electrical activity
• Brief electrical stimulation (5-15 sec) of an
afferent input from the head ganglia causes the
cells to depolarize by 15-20 mV and to generate
an afterdischarge that lasts for about thirty
minutes.
• The nature of the synaptic input is poorly
understood, but it has been suggested that only
a few neurons receive direct afferent input and
that excitation subsequently spreads to the
remainder of the electrically coupled cells
(Mayeri et al., 1979).
• In addition, once the afterdischarge is underway,
peptide release occurs in response to both influx
from the extracellular space and release from
intracellular stores (Arch, 1972; Stuart et al.,
1980; Loechner et al., 1990; Wayne, 1995;
Wayne et al .,1998; Michel & Wayne, 2002).
• In addition ELH acts as hormone that causes the
smooth muscles of the reproductive ducts to
contract and expel the egg string.
• It may also cause the changes in heart and
respiratory beat.
• Simultaneously, the bag cell factor, an inhibitory
transmitter that decreases the rate of specific
neurons (L2, L3, L4, and L6), and beta bag cell
factor, an excitatory transmitter that increases
the firing rate of other neurons (L1 and R1).
•
• The stimulation or inhibition of particular
neurons presumably coordinates the egg laying
behaviour pattern ( Scheller and Axel 1984).
• How does genes fit in to this scenario? The ELH
gene is translated into a large protein that is
broken down into the shorter functional
proteins, ELH, alpha bag factor and beta bag
factor.
• Thus a single gene is activated, and because its
products act as neurotransmitters, and
hormones,
• That gene can orchestrate a complex behavioural
sequence.
• Further more if gene is activated, all the
components of behaviour pattern appear, as a
typical of a Fixed Action Pattern.
• It was the sequence of bases on ELH gene that
revealed the mechanism of this gene’s control of
egg laying behaviour, but sequencing of ELH
gene involved many steps.
• First the ELH gene had to be identified and
cloned.
• To accomplish this an Aplysia DNA library was
created. A DNA library is made up of clones of
different recombinant plasmids containing
fragments of all the DNA of an organism.
• A cDNA library is a collection of cloned cDNA
(complementary DNA) fragments. cDNA is
produced from fully transcribed mRNA found in
the nucleus and therefore contains only the
coding regions of an organism.
• While information in cDNA libraries is a powerful
and useful tool since gene products are readily
identified, the libraries lack information about
enhancers, introns, and other regulatory
elements found in a genomic DNA library.
• cDNA is created from mRNA with the use of an
enzyme known as reverse transcriptase. In
eukaryotic cells, a poly-(A) tail (consisting of a
long sequence of adenine nucleotides)
distinguishes mRNA from tRNA and rRNA and
can therefore be used as a primer site for
reverse transcription.
• Several methods exist for purifying RNA such as
trizol extraction and column purification. Once
mRNA is purified, oligo-dT and random primers
can be used with reverse transcriptase to create
cDNA templates.
• Restriction endonucleases and DNA ligase are
then used to clone the sequences into bacterial
plasmids.
• The cloned bacteria are then selected , commonly
through the use of antibiotic selection. Once
selected, stocks of the bacteria are created which
can later be grown and sequenced to compile the
cDNA library.
• The purpose of a PCR (Polymerase Chain
Reaction) is to make a huge number of copies of
a gene. This is necessary to have enough starting
template for sequencing.
• The cycling reactions :
• There are three major steps in a PCR, which are
repeated for 30 or 40 cycles. This is done on an
automated cycler, which can heat and cool the
tubes with the reaction mixture in a very short
time.
 – Denaturation at 94°C :
• During the denaturation, the double strand melts
open to single stranded DNA, all enzymatic
reactions stop (for example : the extension from
a previous cycle).
– Annealing at 54°C :
•
The primers are jiggling around, caused by the
Brownian motion. Ionic bonds are constantly
formed and broken between the single stranded
primer and the single stranded template.
• The more stable bonds last a little bit longer
(primers that fit exactly) and on that little piece of
double stranded DNA (template and primer), the
polymerase can attach and starts copying the
template. Once there are a few bases built in, the
ionic bond is so strong between the template and
the primer, that it does not break anymore.
– extension at 72°C :
This is the ideal working temperature for the
polymerase. The primers, where there are a few
bases built in, already have a stronger ionic attraction
to the template than the forces breaking these
attractions
• Primers that are on positions with no exact
match, get loose again (because of the higher
temperature) and don't give an extension of the
fragment.
• The bases (complementary to the template) are
coupled to the primer on the 3' side (the
polymerase adds dNTP's from 5' to 3', reading
the template from 3' to 5' side, bases are added
complementary to the template)
•
• Because both strands are copied during PCR,
there is an exponential increase of the number
of copies of the gene. Suppose there is only one
copy of the wanted gene before the cycling
starts, after one cycle, there will be 2 copies,
after two cycles, there will be 4 copies, three
cycles will result in 8 copies and so on.
• Primers sets may be designed using standard
primer design algorithms without any
modification.
• As with all PCR amplifications, however, the
specific reaction conditions for each set must be
optimized, particularly primer concentration,
annealing temperature and magnesium chloride
concentration.
• However, many primer sets fail to amplify the
desired template despite all attempts to optimize
the reaction conditions and a new set of primers
must be designed and synthesized.
• Considering the time and cost of designing and
optimizing primer sets along with the relatively
large number of candidate genes that are
identified by microarray studies, the
development of a central repository for primer
sets, reaction conditions and even the actual
oligonucleotides would benefit all investigators
involved in genomics and other types of
experiments.
• Analysis of Cloned DNA Sequences
• Once DNA has been cloned and amplified, it
needs to be analyzed and characterized. Two
common ways to do this are restriction enzyme
mapping (and its variation, the Southern Blot)
and DNA sequencing, both of which rely upon a
technique known as gel electrophoresis.
• Gel electrophoresis takes advantage of the
overall negative charge possessed by nucleic
acids, due to the phosphates in the backbone.
• Because nucleic acids are negatively charged,
they will migrate in an electric field from the
negative electrode to the positive electrode
(because like charges repel each other).
• If the electric field and the nucleic acids are run
in a semisolid matrix (i.e. a 'gel' usually
composed of agarose, a gelatin-like matrix, or
polyacrylamide, a chemically cross-linked
meshwork), then the nucleid acids will migrate at
a speed inversely proportional to their size.
• In other words, large molecules will migrate
slowly, because they will be held up by the gel,
and small molecules will migrate quickly, because
they can pass more easily through openings in
the meshwork of the gel.
• Practically speaking, a mixture of DNA molecules
can be loaded into a depression in a gel (the
depression is called a 'well'), the gel is
surrounded by an electrolyte solution, and
electric current is passed through the gel.
• The fragments of DNA in the mixture will
separate out according to size, and can be
visualized using a DNA-specific stain, such as
ethidium bromide, which binds to DNA and
glows orange under UV light.
• Once cloned, DNA fragments can be
characterized by restriction enzyme mapping, by
DNA sequencing, or by gene expression studies.
• Next the plasmids bearing ELH gene had to be
identified. This was done by making cDNA from
mRNA in the bag cells and from mRNA in non
neural cells and then labeling each with radio
active marker so that it could be identified.
•
• The cDNA from both sources was hybridized with
Aplysia DNA ineach clone of recombinant
plasmids. The key to hybridization is the
specificity of base pairing.
• Because the bag cells produce ELH, they will
have many mRNA molecules coding for ELH.
• Therefore cDNA from bag cells will have the gene
for ELH. Non neural cells do not produce ELH
and consequently cDNA made from their mRNAs
will lack the ELH gene. As a result the
recombinant plasmid that hybridized with cDNA
fro m bag cells only and not from other cells.
• The sequence of bases in ELH gene revealed that
it codes for a large poly-protein precursor that is
chopped in to several functional proteins.
• The sequence of bases on ELH gene codes for
271 amino acids, but ELH itself has only 36
amino acids.
• A single gene codes for a large protein that is
broken down at least three smaller active
proteins with different roles in producing a
behaviour pattern.
• The role of the genes in generating the this FAP
is to produce peptides that act locally as as neuro
transmitters or distantly as hormones.
• These peptides stimulate or inhibit specific
structures to coordinate a stereotyped behaviour
pattern.