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PHEROMONES: ECTO-HORMONES
Chemical messengers that are transported outside the body that have the potential
to evoke physiological or behavioral changes in another individual of the same species.

Types of pheromones:
1. Signalers: cause short term behavioral changes. (mating signal,
aggression, lordosis).

2. Primers: cause physiological change at the long run. Affect mainly

HPG axis.

While most mammals have VNO in addition to the olfactory organ, humans
Lack it.

Some groups claim that there is a functional VNO in humans:

Vomeropherine (PPD- pregna-4,20-diene-3,6-dione) direct introduction into the VNO
had changed FSH/LH pulses pattern in humans.

Berliner et al. 1996

PHEROMONES ACT MAINELY THROUGH THE VNO-AOB BUT ALSO

THROUGH THE MO SYSTEM.

IN THE END OF THE ROAD THE HYPOTHALAMUS GETS SIGNALS FROM

BOTH ORIGINS.
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flank marking

Mating behavior

hamster

rat male

Plural breeder

Singular breeders

Rodents, humans

Marmosets

The male effect

ewe

Social and reproductive strategies:

Plural breeding VS. Singular breeding

Rats, Humans

Marmosets

In some plural groups synchronization of

menstrual/estrous cycle have been observed:

Rodents:
The Lee-Boot effect (Van der Lee & Boot, 1955, Acta Physiol. Pharmacol.
Neerl. 4: 442)
Grouping several female mice in a cage results in suppression or
modification of their
estrus cycle (delay of puberty and suppression of estrus; effect is female
to female)
The Whitten effect (Adv. Reprod. Physiol. 1: 155)
involves (1) estrus induction, (2) estrus synchronization, and (3)
suppression of estrus in females housed in large group.
The induction of synchronized estrus by urinary cues of conspecific male
mice in females with group-dependent estrus suppression.
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Plural Breeders:

Humans:
The most known phenomenon between females is the :

Cycling Synchronization Effect

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The coupled oscillator mechanism

Ovulatory pheromone lengthens cycle, and
follicular pheromone shortens cycle (Stern &
McClintock, 1998).

(0-2 days
after LH
surge)

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Ovulatory phase pheromones lengthen

Follicular stage

Follicular phase pheromones shorten

Follicular and luteal phases.

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In rats: All female environment tends to suppress the estrus c

Extension of estrus cycle

Induction of spontaneous
pseudopregnancy

Prolongation of quiescent period

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However, although the phenomenon are well described, the neuroend

correlates of female to female effect in plural breeding group species
been published.

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Singular breeders:
Reproduction is limited to a single female in a group.
In this case, a dominant female inhibits the sexual behavior of
subordinate Females. Oocytes are arrested in follicular stage.
Marmosets- primates with developed VNO-

Subordinate female has low LH levels comparing to dominant female.

GnRH levels and pulsatility seems to be unchanged.

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* The effect is not GnRH dependent, what then is the mediator?

* Stress is one of the key factors, ACTH and glucocorticoids
levels are high in subordinate female.
* Opioids, released in response to stress, mediate the
suppression of
LH in subordinate females. Naloxone prevented the LH
decrease.
* In another experiment it was shown that subordinate
females have demonstrated enhanced sensitivity to the
inhibitory influences of E feedback on LH secretion.

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Proven to be sufficient

Cues from dominant female exert 1. increased sensitivity to E negative feed

2. Inhibition of LH secretion from the pit

Both lead to hypogonadotropism

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Male and female pheromonal interaction

Humans:

Pheromonal related neuroendocrine changes in humans brain haven

well studied.

Pheromonal effect on mood and perception in females revealed that

androstenone is perceived as a hedonic scent and that it improves m
of the recipient female just before and during ovulation, e.g. there is
in sensitivity to male pheromone depending on the menstrual stage.

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Animal model

1. The Whitten effect- Induction and synchronization of estrous (ovula

among unisexually grouped females in the presence of a male.

2. The Vandenbergh effect- (Vandenbergh, 1969, Endocrinol. 84: 658)

Onset of puberty in young female mice can be accelerated by phero
contained in the urine of adult males .

3. The male effect- enhancement of ovulation during anestrous with t

appearance of a male.
4. Female urine causes a rapid induction of LH release and mounting
behavior in male.

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Females mice, exposed to male urine or isolated pheromones, show h

estrous cycle frequency (especially in groups).

Jemilio et al. 1986

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The Flank marking behavior of the Hamster

Scent marking behavior. The hamster rubs its flank
against
an object to spread pheromones.
Gonadal hormone dependent type of behavior
Used to communicate a variety of social information
relevant to reproduction and agonistic behavior.
Males- castration reduces the frequency of flank
marking, T restores it.
Females- Frequency correlates with the estrous cycle
and levels of E and P during the cycle. (Female mice
frequency of urine scent marking is not correlated
with changing levels of gonadal hormones).

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Number of flank marks are more frequent in diestrus2 and proestru

and low in diestrus1 and estrus stages.

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E replacement to OVX females

restores cyclicity of flank marks

Progesterone addition inhibits

the stimulatory effect of E

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Flank marking behavior is driven by the activity of

arginine- vasopressin in the mPOA

AVP injection to the mPOA- and only AVP- induces number of flank mar
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V1R antagonist inhibits the AVP induction effect.

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* Female hamsters, implanted with E capsules marked more in res

to AVP comparing to untreated females. i.e. Estradiol influences fl

marking by altering the sensitivity or responsiveness of the mPOA

AVP.

* c-FOS activation studies of flank marking behavior pointed to the

CAN (central amygdala nucleus) and PAG as part of the neural circ

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Based on observation in rats and mice E induces number

of AVP-R (AVP 1aR) (not yet in hamster).

Rat mPOA

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Summary:
1. Flank marking in the hamster female is a way to pheromonally
inform the conspecific male about her sexual status.

2. High levels of E, which occur in late diestrus and proestrus, induc

flank marking frequency.
3. AVP stimulates flank marking frequency by acting in the mPOA.
4. Possibly, E induces AVPaR1 receptor synthesis in the mPOA and
sensitizes the mPOA to AVP.

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The male effect:

similar to Whitten effect but in this case, females

are in anestrus. Exposure of female to male or his odor (fleece) out
of the breeding season activates LH secretion and synchronization
of ovulation. Best known in sheep and goats.

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The odor signal in ewe is perceived by the MO system rather than

VNO.

When chemical anosmia was

applied:

Considerably less females

Responded to male odor.
LH pulses frequency,
amplitudes
and levels were low
comparing to control.

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Further support for the

MO tract of the male
effect was obtained by
temporal local
inactivation of the
Cortical nucleus of the
Amygdala or the VMN by
Lidocaine.

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Only inactivation of the

cortical nucleus of the
Amygdala impaired
LH increase in response
to male.

Gelez & Fabre-Nys, 2004.

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The role of GnRH in pheromonal sensing:

So far, it was shown that sexual related phermone signal results in

an increase in LH pulasatility and circulation levels in females.

Although not shown here yet, the same phenomenon is observed in

But, is GnRH involved in this activation?

(Recall that GnRH is not involved in the hypogonadotropism of marmoset subordinate

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An attempt to answer whether GnRH mediates LH elevation in male mice wh

introduced to female odor

90 min
45 min

GnRH expression in the POA

did not increase when exposed
to female odor. (We already now that
LH levels increase).
*However,GnRH was upregulated in
response to an intruder male.
Similar results were obtained with
the syrian hamster (Richardson et al.
GCE, 2004)

Bedding

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But mating behavior is induced by ICV injection of LHRH

to VNX males. (LHRH is in the circulation and can be transported to
many regions in the brain).

How does GnRH mediate the detection of female odor and male sexual beha
Is there another source of GnRH in the brain?

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Fernandez-Fewell& Meredith, Physiol and Behav, 1995

The Terminal nerve:

* Is a complex of neurons and nerve
fibers that extends from the nasal
cavity to the brain along the course
of the vomeronasal nerves .
* It by-passes the AOB and enters
the forebrain behind the olfactory
bulbs.
* It follows the route by which the
precursors of all forebrain GnRH
(LHRH) neurons migrate into the
brain during embryonic
development, and remains in
adults as an additional neural
connection from nose to brain that
is rich in GnRH containing cells and
fibers.
* The terminal nerve is present in
all vertebrates whether or not they
have a VNO and it has been
suggested that the nerve fibers
apparently innervating the human
VNO are in fact NT fibers.

NT GnRH neuron terminals ends in the

median eminence.

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GnRH neurons in the NT project

centrally into the ventral forebrain
and periphally into the nasal
chemosensory mucosa.

GnRH receptors are exspressed

by chemosensory neurons in
VNO and MOE.

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GnRH alters the response of chemosensory neurons to stimulation by

modulation of voltage/receptor gated channels.

Using patch clamp technique,

GnRH bath increases sodium
coductances in olfactory
neurones

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Hypothetical model for GnRH modulation

Neural and hormonal inputs

control the amount of GnRH
available in and secreted by NT GnRH
neurons.
*GnRH diffuses and influences
the response of chemosensory neurons
to odor
*Stimulation of GnRH-R activates second
Messenger pathways that alter the
odor-induced membrane potential and the
signals to be sent to the brain.
*However, the link between NT GnRH and LH
change hasnt been shown yet.

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Terminal nerve damage impairs the mating behavior of the

male hamster

The results are not really convincing..

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Take home messages:

Pheromones have a role in supporting reproduction, thus increasing
the chances for successful mating.

Pheromones are not the only factors. Other sensory factors,

memory and learning co-integrate to ensure the success of reproduc

Pheromones work in accordance with the type of social organization.

The mechanism by which LH secretion is affected is not fully underst

in many of the cases I presented today.

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